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Prostaglandin F2α disappearance and concentration in amniotic fluid and blood of women undergoing abortion with intra-amniotic hypertonic saline
PROSTAGLANDIN P2 DI S A P P E A NCE AND CONCERT TION A I O r l C Y L 0 ~ D A N D B h O O D OF W O M E N U N D E R G O I N G a B O R T iO N W ITH I a I O T I C H Y P E R T O N IC S A L I N E
C.Y. P U R l N. B A L I G A , N. V . HIN130 N1 AND K . R .
AGGAR L LA S
De p a r treen t o f R p r o d u c t x4 v e ,, B i o l o g y a n d the o f Ob s te t r ics a n d G y n a e c o i o gy All
IN
Department
I n d i a I n s t i t u t e of M e d i c a l S c i e n c e s New D e l h i - I f 0 0 1 6 , iNDIA
ABST
CT
Int ra- a m n £ o t ic a d m i n i s t ra t i o n o f 2 0 p e rc e n t h y p e r t o n i c s a l i n e c a u s e ~~',~ a s lo g n mif i c a n t increase in t h e c o n c e n t r a t i o n o P G F 2 a in the a m n i o t i c fluid o£ women undergoing second
trimester a b o r t i oQn • The increase ie n PGF2a w a s s l o w i n " .e patients with longer abortion tim Interestingly, x"n o n e case (GS), the levels o f PGF a i n c r e a s e d from 0.32 to 7.48 ng/ml w n the induction-abo tion time was 7.75 hours. H o w e v e r , no s i g n i f i c a n t c h a n g e in P G F z a l e v e l s w a s o b s e r v e d in the p e r i p h e r a l v e n o u s b l o o d d u r i n ~ i ~ d u c t i o n - a b o r t i o n interval. T h e d i s a p p e a r a n c e o f (96--~H)-PGF ~ a n d its rectah o l l i e s w a s s l o w in t h e a ~ n i o t i c f l u i d of s l i n e ad "nist e r e d c a s e s as c o a r e d w i t h the c a s e s w h e r e s a l i n e w a s not g i v e n . T h e s l o w d i s a p p e a r a n c e o f P GF2a a n d its m e t a b o l i t e s in the s a l i n e a d m i n i s t e r e d a m n i o t i c f l u i d h a s b e e n u g g e s t e d to be p a r t l y due to t e r a p i d c h a n g e in a iotic uid volume a~d increased synthesis/release o f P G F 2 a in the a iotic fluid. F r o m the a m n i o t i c f l u i d , P G P 2 a w a s diff u s e d i n t o the f o e t u s w h e r e v a r i a b l e a m o u n Z s w e r e o b s e r v e d in d i f f e t e n t t i s s u e s. The s e o b se rva t io n s sug g e s te d that f o l ! o w i n g the a d m i n i s t r a t i o n of hypertonic saline, an i nc re a s e d e n d o g e n o u s s y n t h e s i s / r e i e a s e o f P GF 2a in he a m n i o t i c f l u i d w a s o b s e r v e d w h i c h m a y a c t o n the u t e r u s l o c a l l y a n d c o n t r i b u t e to the a b o r t i f a c i e n t p r o p e r t i e s o f 20 p e r C e n t h y p e r t on i c s a I i ne.
Accept
e
u for
DECEMBER
pubiication
1975
VOL.
~epte~ber
12 NO.
6
~0,
1975
679
CONTRACE
ION
INTRODUCT ION Since the eariy 1960s intra-uterine administration of hypertonic saline has been widely used for the t e r m i n a t i o n o f m i d t r i m e s t e r p r e g n a n cy. W h e n c a r e f u I Iy and has a a d m i n is t e r e d , t h i s m e t h o d is h i g h l y s u c c e s s f u l A multitude I o w mo r b i d i t y r a t e w i t h f e w s i d e e f f e c t s . fall, foetai death, o f p o s s i b le f a c t o r s l i k e p r o g e s t e r o n e i n c r e a s e d i n t r a - u t e r in e p re s s u r e a n d o x y t o c in- I ike a c t i o n h a v e b e e n p t o p o s e d t o e i u c i d a t e t h e ah o r t i f a c i e n t ac t i o n There are varying reports on the o f h y p e r t o n i c s a l i n e [i). saline on serum hormone levels. A effect of hypertonic few s u g g e s t e d t h a t h y p e r t o n i c s a l i n e l o w e r e d s e r u m p r o ievels (2), w h i l e o t h e r s do n o t f i n d d e p l e t i o n gesterone I n o u r e a r 1 i e r s t u d i e s (4), in p r o ge s t e r o n e I e v e I s [ 3 ) . a significant decline in serum progesterone w e d i d no t f i n d a n d human p i a c e n t a l l a c t o g e n levels after i n t r a - a iotic administration of hypertonic saline, whereas PGF~= and 15(S)15-methyl-PGP2~ produced a significant f a l i in t h e i e v e i s of these hormohes during the induction-abortion interval. Extensive increase in i n t r a - u t e r i n e volume by intra-amniotic administration o f i s o t o n i c s a l i n e f a i l e d to i n d u c e a b o r t i o n (5). G u s t a v i i ( 1 ) s ug g e s t e d t h a t t he ab o r t i f a c i e n t a c t i o n of hypertonic saline was probably d u e to t h e d e g e n e r a t i o n of decidual cells. P r e i i m i n a r y r ep o r t s w i t h e x t r a - a i o t i c h y p e r t on i c saline have sho i n c r e a s e s in P G F 2 ~ l e v e ! s in t h e a m n i o t i c f l u i d (6,7). However, t h e r e is n o s y s t e m a t i c s t u d y to v e r i f y this Inc ease after intra-amniotic administration o f 200 m l o f 20 p e r c e n t h y p e r t o n i c sa!ine. It was with this aim that the disappearance of ( 9 $ - 3 H ) - P G F 2 a , a n d its m e t a b o l i t e s in the amniotic fluid of saline admini stered cases have been c o m p a r e d w i t h t h e o n e s in w h i c h s a l ine w a s n o t g i v e n , The levels of PGF2 in the a iotic fluid and blood have been m e a s u r e d a f t e r .t ...h e a inistration of hypertonic saline. The distribution o f 5 H - P G P 2 ~ in t h e f o e t a l t i s s u e s h a v e a I s o b e e n i n v e s t i g a t e d. A r e i i m in a r y r e p o r t h a s b e en p r e s e n t e d (8). T E R IAL S A N D
METHO D S
Sub ~ects
T w e n t y - t h r e e h e a I t h y w o m e n , b e t w e en 16 - 20 w e e k s g e s t a t ion, s e e k i n g m e d i c a l t e r m i n a t io11 o£ p r e g n a n c y were randomly selected for the present studies, in t h i r t e e n cases, pregnancy was terminated with intra-a iotic PGF2a w h e r e a s t h e o t h e r t e n w e r e g i v e n 200 ml o f 20 p e r c e n t s a i ine int r a-a i o t i c a I ly.
680
CEMBER
1975
VOL.
1 2 N O ....6
CONT 4
In t r a - a
Q
~
I
Q
io t i c a d m i n i s t r a t i o n
of
p
CE
ION
GF 2
With due aseptic precautions, a n n ot was per rmed er abdomen and polyethylene catheter threaded through P h e 1 8 - g a u g e n e e d l e as m e n t i o n e d in d e t a i l e a r l i ~ ~ (~)H" F i r s t d o s e o f 25 m E P G F 2 ~ w i t h 50 to 50 ~ Ci o f ( - ) P GF~was injected throug~ this catheter and t second dose of only radioinert PGF2~ was given after six hours. Q
t
~
I.
.
.
.
3
(gB-SH]-PGF2 630 to 50 :aCi), s p e c i f i c a c t i v i t y 7.5 ~ C i / ~ m o i e, in 5 ~ I s a I ine w a s i~ j e c t e d i n t o t h e a m n i o t i c f l u i d in f o u r w o m e ~ i2 h o u r s p r i o r to t h e i n j e c t i o n of 25 m g P G F 2 a . I n t r a - a n n i o t.... i", . '::..a ......d ..... m.......i ...... n......i ...... s...... t...... r....a.......t ......i ..... o....n.............o.......f...........h ..................e ......r ......t ......o...... n......i.....c ............s......a......I......i.....n ..... e................... ~,o
i
|
200 Millilitre o f 20 p e r c e n t s a l i n e w a s a d m i n i s t e r e d intra-a i o t i c a l l y o v e r a p e r i o d o f a b o u t 20 m i n u t e s in six cases. I ediately following t h a t , 30 to 50 ~ C i o £ (gs- ~ H ) - P G F 2 ~ w a s in e c t e d i n t o t h e a iotic flui through the sa~e catheter.
lntr
a
a
i o t ic
...........................
admin i s t r
-,.
o:f _~h y p e
on "
/ ...................................................... ...........: ...........................................................................................................................
As ~entioned a b o v e , 200 m l o f h y p e r t o n i c in j e c t e d i n t ra- a i o t i c a I 1y i n a n o t h e r S r o u p PGF2~ was not injected in t h i s g r o u p . Co i i e c t ion the
a
o f b Ioo d and
n i o t i¢ f l u i d inistration
a n n io t i c
saline o f fi
was women.
£i u i d
a n d b 1 o o d s a m p I e s w e r e co 1 le c t e d b e fo re of hypertonic s a l i n e or P G F 2 a a n d t h e r e -
after long
at regular i~tervals as free fiow of liquor c a t h e t e r. Ca 1 c u l a t i o n
..............................
o £ ha lf-I
till was
the patient mainta ~ed
abSrted through
or the
so
i £e
To calculate t h e h a l f - ire t i m e o f ( - S H ) ....P G F 2 and its metabolites in t h e a m n i o t i c fluid, the best fit i ines were dra by the me thod o £ two var lab I e I i~ear D e p e n d "i n g u p n t h e p a t t e r n o f d "i s a p e a r a n t e, re g r e s s i o n. single line was dra in cases where linear pattern was observed a n d t w o l i n e s in c a s e s w h e r e b i p h a s i c p a t t e r n fo i 1 o w e d. I ~ e a c h c a s e, t he c o r r e I a t i o n c o e f f i c i e n t w a s greater than 0.95. Pro~ the slope of each line the first ( a t l / 2 ) a n d s e c o n d ( S t l / 2 ) h a l f - i f ' y e s w e re c a l c u l a t e d .
DECEMBER
1975
VOL.
12 N O . 6
681
C
TR
Uvtake
E
+I+ON
o£
and
it s m e t
ol ite s
S Ue S
P o e tus
wa s
co 1i e C te d
a long
w i~ t ~h.
p. I a. c e.n t a
ii
e di a t e ly
after the patient aborted with the use of radioinert and 5H-PGF2a t It w a s t h o r o u g washed with chilled saline and d isse c te d ou t to co i Iec t a I E i s s u e s, A d r e n a i, T a in j k i d ~ e y , i i v e r, I u n g s, m u s c le, s k n p i acen t a and u~b i i i c a 1 cord were minced, washed, dissolve~ in s o i u e n e a n d c o u n t e d in t o l u e n e b a s e d s c i n t i l l a t o r . ~n t w o c a s e s , f o e t a l t i s s u e s re e x t r a c t e d w i t h ~ e t h a n o l : c h l o r o f o r m and the dried e x t r ac t
c o u n t e d. f _ .......r a
ioact
T r it i con t ent s m e a s u r e d in t h e a i i q u o t s o £ s e r and a iotic fluid s i t s in d u p l i c a t e u s i n g t o l u e n e ...b .ased scintillator with biosolv ( B e c k m a n ) as w a t e r e m u l s i f l e r ~n a N u c l e a r C h i c a g o l i q u i d sc i n t i l l a t z"o n c o te r Mark If. Remaining s les w e r e ~ r o z e n in a l i q u o t s b e l o w - 1 5 ° C . Ex r
ct..ion........a......n.....d
chroma
gr
h y ....
P G F 2 a w a s e x t r a c t e d f r o m a ~ n £ o t ic £ 1 u i d b y t h e m e tho d o C a I d w e i I e t a 1. (9). In b r iv f, n i o t ic f i ui d w a s r e n d e r e d acldlc to 4 . 0 ) w i t h O . 1 N H C I a n d e x t r a c t e d w i t h
diethxl
The
chromatographed
e~tract
was
washed
with
water,
dried
and
through
s£1ic£c acxd col ns u s i n g i n c r e a s i n g ~ethanol in b e n z e n e :e t h y l a c e t a.t.e. . . . ( 3; 2 -) .
concentrations of Eluted PGF2a was dried
and
f urther
subjected
t h in
to
1
er
chromatography in c h l o r o f o r m : t h a n o l :a c e t i c a c l d : w a t e r (90:9 : I:0.6) system (I0). PGF2a zone parallel to t h a t o f a u t h e n t i c P G F 2 a w a s e x t r a c t e d a n d co ted £or he t r i t i conte~t. Ra~ £o
no
s
PGF 2 a
0.2 to 12-ng PGF~. To r u n t h e a s s a y , about ~ 500 cp~ of 3H-PGF,~ was added to eacfi sample and then ~extracted and chro~atograPhed ~ through sil£cic acid colu s (9). The ¢luant was dried a~d made to one ml With ethanol. An a l i q u o t was taken each sa to account f o ~ ... r e c o v e r y , which fro~.65 t o 80 percent. ~he remaining s le s dried a n d ~ a d e t o O. I m l with 0.01 M Tris-HCl buffer (pH 7.4) containing 0.15 M NaCl and 0.! p e r c e n t g e l a t i n . 0.I MI diluted 1 / 4 0 0 g a v e a b o u t 55 p e r c e n t .binding)
tubes. ature
682
The for
t
es
were
30 m i n u t e s .
ently ~H_PGF vo rtexed 2~
(8000
antgser was added
and cpm)
~DEcEMBER
left
(a d i l ~ t i o n o f to all t h e a s s a y
at
-in 0.i
1975
r o o m ~te ml
buffer
VOL.
12
was
. 6
C
T R
E
I
a d d e d to all the tubes and the r e a c t i o n m i x t u r e i" n c u b~,ated at 37"C for one hour. This w a s ! l o w e d b y the a d d i t i o n o £ 0 . 1 ml o£ 1 / 1 0 0 n o r m a l r a b b l t s e r u m a~d t h e n 0.I ml o£ g o a t a n t l s e ~ to r a b b i t ga a globulin. The 01e r e a c t i o n ~ixture was left ove rnight
a~
Z to
4 "C
and
c e nt r i fuged
at
55 O D rp~
£or
40
m £ n u t es.
The pre~ :ipitate w a s d i s s o l v e d :in 0 2 ml o£ 0 I N N a O H and c o u n t e d c. To t the s e n s i t i v l t y a"n d r r o .c i b i l i t y of the a s s a y m e t h o d , d i f f e r e n t c o n c e n t r a t i o n s of r a d l o i n e ~ t PGP2a w e r e a d d e d to 3 mi of s e r u m oT ioric f l u i d and the s les extrac~ed, ¢ h T o m a t o g r a p h e d and a s s a y e d as d e s c r i b e d above. Nonspeciflc b i n d l n g was a b o u t ~ to 2 p e r c e n t en the a n t l s e r was~rep!aced w i t h n o r m a l tab it s e r u m ..... The l e a s t d e t e c t ~ b l e level o f P O P 2 a for l l n e a r l z e d curve, f r o m 0.6 to 1 0 ng~ was 0.8 ng.
RE SULTS earance
~i
of PGF2~ --~::_-
from a
iotic ..........
;::_n
fluid ~-w
~
T w e n t y flve ~ i l l i g r a m s o£ P G F 2 ~ w i t h ( 9 ~ - 3 H ) - P G F z s was a £nistered to five wo~en, o t h e r dose of r a d i o i n e r t 2S mg PCF2~ w a s g i v e n a f t e r six hours. The a m n l o t i c ffluid s a m p l e s were e x t r a c t e d and c h r o m a t o g T a p h e d to a n a l y s e PGP2a c o n t e n t at v a r i o u s t i~tervals. As sho in Fig. i, the r a t e of d i s a p p e a r a n c e of PGF was d i f f e r e n t in d i f f e r e n t cases. In the five cases- AP ~P, C P , D P , E P - 62.5, 38.6, S0.0, 45.8 and 43.8 percent, r e s p e c t i v e l y , P GFz~ d i s a p p e a r e d in 24 hours of P G F 2 a a inistratio~. On an averages 45 8 p e r c e n t PGP2a d i s a p p e a r e d in 24 h o u r s r near the tl of abortion. Dis
p
rance..Of 5 H T P G F 2 ~
and
its m e t a b o l i t e s
from
The dis p e a r a ~ c e o £ 3 H - P G F 2 ~ a~d its m e t a b o l i t e s in the a m n i o t i c £ 1 u l d of r wo~en a i n i s t e r e d w i t h N0 to S0 ~Ci o£ 5 H - P G P 2 ~ is sh n in Fig. 2. In a l l ~he ff r cases, r a p i d d e c l i n e of H - P G F 2 a and its m e t a b o l i t e s was o b s e r v e d to six hour s o£ 3H-p GP 2 a i n i s t r a t ion. T h e a v e r a g e f i r s t hal f- li fe b e i n g 5.3 + 0.7 (me an ÷ S.E.) h o u r s . 3.5 to 6.7 hours). w e v e r , £W two c a s e s , ere a iot£ d sa los c o u l d be c o l l e c t e d up to 12 h o u r s of P G F 2 a a d m l n i s t r a t i o n , he d i s a p p e a r ante off 5 H - P G F 2 ~ and its metaboiites s l o w e d do to a s e c o n d halff-life of 9 ÷ 0.6 hours.
CEMBER
1975
V O L . 12 NO. 6
CONTRACE
ION
A
~k
T
gO"
60. ~CP
,AP
i
G.
.
.
.
.
.
4
.
W:
................................
S
Figure
1.
Dis flu
...........
~ .........
! 0
U
A
S
earance of prostaglandin F from amniotic of wo~en after i n t r a - a m n i o ~ c injection.
(gs-3H)-PGF~a was injected intra-a~niotically along with 5 ~ g P G F 2 ~ in f i v e women. Another 2 5 mg d o s e o ~ PGF2m w a s g i v e n after 6 hours. niotic fluid samples were extracted and chromatographed to deter ine the amount o~ PGF2~ d i s a pp e a r a n c e a t v a r i o u s t i~ e i n t e r v a 1 s.
684
DECEMBER
1975
V O L . 12 N O . 6
NTRACE
laO00
ION
-
l0
8 000-
0
6000-
_u
O
_o
E
0
:~ e ooou
0
2
4
6
8
i0
HOURS
Figure
2.
D i s a p p e a r a n c e of C g B - 3 H ) - p r o s t a g l a n d l n F2a and its m e t a b o l i t e s from nlotlc fluid o£ w~men a ft e r
int ra- a
i ot ic
inj e c t i on.
4
( 9- 3H) -PGF2} wa s ~nj ec t d tnt ra- a i o t .ic a 1 1 ) , in a gro o four women, iotic fluid s les were ana lys ed £o PG .F 2 a a n d i t s m e t a b o i i t e s c o n t e n t a t va r i o u s t ime i n t e rv a I s. Ra d i o i n e r t PGF s not g i"v e n t o t h i s g r o u p of cases
dur
CEMBER
1975
g the course of our study.
VOL.
12 N O .
6
CONTRACE
ION
Dis
O~
e
ante
3H,
2a a
_itS_
_ta_bol
s from
amniotic
T h e dl s a p p e a r a n c e o 3H. P G F ~a. and i t s m e t a b o I i t e s f r o m t h e a m n i o t ic ffluid o f w o m e n a d m in I s t er ed 2 0 0 ml h y p e r t o n i c s a l i n e is s h o in Fig. On t h e s e m i ! o g s c a l e , in 4 of the 6 c a s e s j the d l s a p p e a r a n c e s h o w e d a b i p h a s i c p a t t e r n . The i n i t i a l d l s a p p e a r a n c e o f 3 H . P G F 2 a a n d its m e t a b o l l t e s w a s fast, an a v e r a g e f i r s t h a l f - l i f e b e i n g i 0 . 0 + 2.3 h o u r s (range 3 . 3 to 18.8 h o u r s ) . However, afterwa s, the d i s a p p e a r a n c e was s l o w e d do to an a v e r a g e s e c o n d h a l f ...l. i f e o f 24.2 ÷ 5.8 hours. In two c a s e s (LS, KS), l i n e a r dis t r l b u t ion p a t t e r n w s o b s e r v e d w i t h a h a l f - l i f e o £ IZ.0 a n d 18.8 hours, respectlvelv. and A co a r i s o n of the f i r s t a n d s e c o n d h a l f - l i f e of P G F 2 a its m e t a b o l i t e s in the a m n i o t i c f l u i d of n o r m a l a n d h y p e r t o n i c s a l i n e a d m i n i s t e r e d c a s e s is p r e s e n t e d in T a b l e I . I t c a n be s e e n t h a t the f i r s t h a l f - l i f e w a s i n c r e a s e d f r o m 5.3 + 0.7 h o u r s to 10.0 ÷ 2.3 h o u r s . H o w e v e r , this d i f f e r e n c e was n t s t a t i s t i c a l l y significant. On the o t h e r hand, a s i g n i f i c a n t i n c r e a s e ~p~0.01) in the s e c o n d h a l f - l i f e o f P G P 2 s a n d its metabo.iites w a s o b s e r v e d in c a s e s a d m i n i s t e r e d with hypertonic sallne, in b r i e f , a p r o l o n g a t i o n of the h a l f - l i f e t i m e of P G F 2 ~ a n d its metabolites w a s o b s e r v e d in c a s e s i n j e c t e d w i t h hypertonic s a l i n e PGF2~
levels
in a m n i o t i c
fluid
The levels of PGF2a were measured by a radioi noassay ich h a d a s e n s i t i v i t y o f 0.8 ng. The r e l i a b i l i t y of radioimmunoassays as a t e c h n i q u e ffor the m e a s u r e m e n t of PGP2 le-~els has y e t to be e s t a b l i s h e d (I!). H o w e v e r , the a n t i b o d i e s u s e d in the p r e s e n t s t u d i e s b i n d m o s t e f f e c t i v e l y w i t h the h o m o l o g o u s a n t i g e n - P G P z a . A b o u t 33 n g of P G F 1 330 n g of 1 5 - k e t o - P G P 2 a and m o r e t h a n 1 pg o f 1 5 , 1 4 - d i b ~ d r o ! S - k e t o - P G P 2 a w e r e r e q u i r e d for 50 p e r c e n t i n h i b i t i o n v a l u e s
(12).
The effect of hypertonic saline on PGF2~ levels in the a i o t i ¢ f l u i d is sho in Fig. 4. The l e v e l s of P G F 2 a s t a r t e d i n c r e a s i n g a f t e r the a inistration of s a l i n e . In t h r e e c a s e s (HS, IS, JS), w i t h an a v e r a g e i n d u c t i o n - a b o r t i o n time of 2 8 . 7 5 h o u r s , i n c r e a s e in P G F 2 s l e v e l s s t a r t e d a f t e r 8 - I 0 h o u r s o f saline administration, a n d n e a r the t i m e of a b o r t i o n , a s i g n i f i c a n t i n c r e a s e in P G P ~ a l e v e l s h a d o c c u r r e d In one c a s e (GS), w h e r e p r e g n a n c y e r m i n a t e d aft 7.75 h o u r s of saline a i n i s t r a t i o n , a s h a r p i n c r e a s e in P G F z a l e v e l s w a s observed. I n this case, P G P 2 a l e v e l s i n c r e a s e ~ f r o m 0.32 t o 7.48 n g / m l a m n i o t i c fluid.
1
DECEMBER
1975
L. 12 N O . 6
I0
COS) ~ t
=6"3HR t
A
~N ~ME
HR
i ;i
OS 5 J
6
.
.
.
.
.
.
ii!~i~
30.1
g 4
MS
Z
~S 0
J5
4
2
6
8
i0
12
!4
16
18
20
22
26
HOURS
~£guTe
3.
Dis i t s
pearance of f9g-3H)-prostaglandin ~ e t a b .o i i t e s £ r o ~ hyp e r t on I c s a I I ne
tered
a
iotlc
fluid
of women
F a
and !ni s -
undergoi:ng abortlon.
3 ...... p C9~-H ) GF2 was injected i d l a t e i....y a f t e r t h e a inlstrat[on o 200 m l off 20 p e r c e n t s a l l n e in s Ix wo~en • i o t Ic f l u i s [ e s wet e iys ed to d e t e £ n e t h e d l s a p p e a r a n c e o £ P G F 2 a a n d Its me t ab o i i t e s a t v a t i o u s t e int e a is 2 Rad io I n e r t P G F 2 a w a s n o t g l v e n to t h e s e c a s e s .
DECEMBER
1975
VOL.
12 NO.
6
687
b~
)
~a
b
N.S.
[0.0 + 2.3 (6)
~.3 + 0.7 (4)@
p¢O.O1
24.2 + 5.8 (3) #
9.7 + 0.6 (2)
HOURS
(MEAN + s.E)
(ME.~ + S.E) HOURS
#
In two cases [LS and KS), linear distribution of (gs-bH)-PGF2a and its metabolltes wis observed with a half-hfe of 12.0 and 18.8 hours, whereas in case (OS) sam phng ' was not possible after 6 hours.
@ The figure in parentheses is the number of observations.
** Disappearance of [98-3H)-PGF2a and its metabolites from amniotic fluid of women administered with intra-amniotic hy~ertonic saline.
* Disappearance of {98-3H)-PGF2a and its metabolites from amniotic fluid of cases where saline was not glve " n.
Level of
~ypertonic saline** tdministered cases
~orma] cases*
~ASES
SECOND HALF-LIFE
;!RST H_ALF-LIFE
EFFECT OF HYPERTONIC SALINE ON DALF-LIFEOF (! -PROSTAGLANDIN F2a AND ITS METABOLITES IN .~h?;IOTIC FLUID
tABLE I
Z
m
,q
>
3Z
CONTRACE
ION
GS A
:Y
:J~E
N$
4
8
i2
~6 H
Figure
t
0
u
R
s
Prostaglandin
F2a levels in amniotic fluid o f ,,,omen a d m i n i s t e r e d w i t h £ntra a m n i o t i c h y p e r tonic s a l i n e for t e r m i n a t i o n of p r e g n a n c y .
P G F Z a i e v e I s w e t e me a s u r e d b y r a d i o i noa s say. Ea c ~ s p 1 e wa s e x t r a ¢ t e d a ¢ d c h r o ~ a t o g r a p h e d through silicic acid columns and assayed using a n t i s e r u m r a i s e d a g a i n s t PGF coup led to po Iy£-lysine in rabbits and (9B)-PGF2a (about 7 pmole) as tracero The antibodies used in this study bind st effectively 'ath the homologous a n t i g e n - P GF 2 a -
DECEMBER
1975
VOL.
12 NO. 6
CONTR
EPTION
Is i : : b o d
2
The effect of h ertonic saline on serum levels of PGF2a in five w o m e n u n d e r g o l n g t e r m l n a t i o n of p r e g n a n c y is s h o w n in T a b l e II. No c l e a r p a t t e r n of s e r u m l e v e l s of p G F 2 , c o u l d be o b s e r v e d , in one c a s e (GS), the s e r u m levels o£ P G ~ a be for (l. Z0 n g / m l ) and a £ t e r six h o u r s of s a l l n e a d m i n l s tration (0.95 ng/ml) were more than observed in other cases. The £ n d u c t i o n - a b o r t l n - l n t e r v a l , 7.75 hours, was the l o w e s t in this case. D:istribution
of
3H
2a
and
its
:metaboIites
The di'stribution of 3 H - P G P 2 and its m e t a b o l i t e s in the various foetal compartments, a~ter intra-amniotic a inistration of a s i n g l e dose o£ 40 m g F2 , is s h o i"n T a b l e III. PGF 2~ a n d its m e t a b o l i t e s w e r e f o u a i s t r i b u t e d in d i £ f e r e n t a m o u n t s in a11 the t i s s u e s i n v e s t i g a t e d . The u p t a k e was maxl m in the u m b i i i c a l cord. S k i n and m u s c l e also h a d m o r e radioactivi .... t h a n o t h e r tissues. In three of the £our cases, the u p t a k e ot~ 5 H . P G F 2 a a n d . i t s m e t a b o l i t e s was m o r e in a d r e n a l th in l i v e . T h e s e p r e l l m i n a r y s t u d i e s also s h o w e d a t r e n d which indicated that there was a somewhat higher take of PGP a a n d i t s m e t a b o l i t e s when the foetus was exposed to PGF a for a l o n g e r p e r i o d of time, as s h o w n in case 4.
DISCUSSION The p r e s e n t i n v e s t i g a t i o n s on the m o d e of a c t i o n of i n t r a - a m R i o t i c h y p e r t o n i c s a l i n e for the t e r m i n a t i o n of s e c o n d t r i m e s t e r p r e g n a n c y s u g g e s t e d that 20 p e r c e n t saline gradually increased the levels of PGF2a in the amn £ o t flu i d a s t he t ime o £ ab o r t ion a p p roa ch e d. The d e l a y in he e l i m i n a t l o n o f P G F 2 a a n d i metabolites f r o m the ...a..m n i o t l c f l u i d o f w o m e n a i n i s t e r e d w i t h hypert o n i c s a l i n e also i n d l c a t d the p o s s i b i l l t y of i n c r e a s e d of P G F 2 ~ in t h e a iotlc fluid. observations along with t e e s u l t s ....that ...no . s gnificant c h a n g e in p e r i p h e r a l s e t l e v e l s o£ P G P 2 a w a s . o b s e r v e d d u r i n g the i n d u c t i o n - a b o r t l o n interval s u g g e s t e d that r e l e a s e d i n the a iotic~fluid y on ..the .. Ut
rU
locally
pr p e r t i e s
of
and
contribute
20 p e r c e n t
to
the
ab0rtifacient
saiine.
The h a l f life of PGF2a in longer than in blood. ...... A b o u was l e f t in c i r c u l a t i o n
the a iotic fluid is much 3 percent ....o injected PGF2 o n e and a h a l f m i n u t e o f f
DECEMBER
1975
L .... 1 2 N O .
6
""~'~
7.75 34.50 25.25 28.50
IS
~IS
IS
;S
0.32
0,65
0.48
1.20
0 Hr
0t.27
0.61
0.24
0,95
6 Hr
0.41
0.75
0.55
-
12 Hr
0.51
0,89
0.90
18 Hr
0.46
0.78
0.71
24 Hr
PROSTAGLANDIN F2a LEVELS ....................... ng/ml ........................... _
IN BLOOD OF W O R N ADMINISTE~D WITH INT~-A_~NIOTIC SALINE FOR TERMINATION OF PREGNANCY
PGF l o"els were measured by radioimmunoassay, The v a l u e s have been exp as ng/ml s e r ~ a f t e r necessary c o r r e c t i o n for losses incurred during e x t r a c t i o n and chromatograph)'.
ABORTIONINTERVAL Hr
CASE
INDUCTION-
PROSTAGLb.NDIN F, 1
rABLE II
,q
.4
3,720* " 20,000 ~,I00 1,580 2,580 2,900 4,620 8,410 ~
ADRENAL
PLACENTA
LIMBILICA] CORD
SKIN
BRAIN
I@JSCLE
LUNGS
KIDNEY
LIVER
4,090
4,900
2,520
6,220
-
5,190
16,020
930
5,600
FOETUS 2 (IAT 19.5 Hr)
5,220
6,550
5,400
5,160
1,450
8,520
"
1,160
7,290
FOETUS 3 (IAT 31.5 Hr)
11,700
4,500
19,020
1,650
18,870
23,060
3,120
12,390
FOETUS 4 (I~:T 34.0 Hr)
TISSUES AFTER !NTRA-~IOTIC )STAGLANDIN F2=
*~. . . . . . . ~ are expressed as d i s i n t e g r a t i o n s per minute per gm t i s s u e . (! ~ (40 ~Ci) was administered i n f o u r women undergoing t e r m i n a t i o n of pregnancy w l t h i n t r a - a ~ i o t i c "~ "40 mg). Foetus and p l a c e n t a were collected inediatel7 after the aborted. Foetal t i"s s u e S were washed, minced and counted for the t r i t i u m content.
FOETUS I (IAT 13.0 Hr)
TISSUES
JPTAKE OF P~OSTAGLkNDIN F2o AND ITS METABOLITES BY ' ~MINIST~TION OF RADIOINERT AND (gB-
tABLE I I I
Z
),
Z
CONTR
E
ION
its administration (15). The significant increase (p~O.O1) in the second half-life o f PGP2~ a n d i t s m e t a b o l i t e s in s a l i n e a d m i n i s t e r e d a m n i o t i c f l u i d as c o m p a r e d w i t h c a s e s in W h x~ c h s a l i n e w a s n o t given, c o u l d at l e a s t be due to c h a n g e in a m n i o t l c f l u i d v o l u m e a f t e r the a d m i n i s t r a t i o n o f 20 p e r c e n t s a l i n e a n d i n c r e a s e d e n d o g e n o u s s y n t h e s i s / r e l e a s e of P G P 2 s in the a m n i o t i c f l u i d . S i n c e the a d m i n i s t r a t i o n o f 40 g~ s o d i u ~ c h l o ide p r o d u c e s i o n i c i m b a ! a n c e in a i o t i c fluld, the b o d y f l u l d s e n t e r the a iotic 1me . c a v i ty c au s i ng a c h . a n.g e. in . . a m.n i.o t.l c . £.1 u.l d . v.o l u . m e. w i t h t" It
is
possible
that
the
change
in
volume
af
cted
the
ellmin ion p a t t e r n of P G F 2 ~ and its m e t a b o l i t e s . Brewer et al. ( i ) h a v e r e p o r t e d ~a net i n c r e a s e in niotic fluid 1 assoclated with a decreased blood volume within 2 to 4 h o u r s f o l l o w i n g i n t r a - a m n i o t i c i n e c t i o n of h y p e r t o n l c sallne. Subseque 1y, the a m n i o t i ¢ fl~ id v o l u m e s a r t e d d e c l i n i n g and W i t h i n "8 ~to I0 h o u r s , r e t u r n e d to n o r m a l levels. It has a l s o b e e n r e p o r t e d t h a t the h a l £ - l i f e o£ d e u t e r ium o x i d e i s I e s s in o I igo - h y d r a i o s t h a n in p o lyh y d r a m n i o s ~159. It thus a p p e a r s t h a t in a u d i t i o n other factors, endogenous synthesis/release Of PGF 2 and c h a n g e in the v o l u ~ e o f a m n i o t i c .fluid m a y c o n t r i b u t e to t h e s l o w d i s a p p e a r a n c e of P G F 2 ~ a n d its m e t a b o l i es in the a m n i o t i c f l u i d o£ s a l i n e a i n i s t e r e d cases. F u r t h e r m o r e , C s a p o (16) s u g g e s t e d that the i n c r e a s e in niotic f iuid volume after hy.pertonic s a l i n e t r e a t m e n t f a c i l i t a t e s a b o r t i o n b y e n h a n c i n g uterine v S i n c e it is k n o ~ that p r o s t a g l a n d i n s a r e r e I e a s e d i n r e s p o n s e t o u t e r i n e s t r e t c h , c e 1 i u i a r dama and
to
vaginal
enhanced in P G P 2 s
distention
(17,18),
i t ~is p o s s i b l e
uterine activity may also contribute c o n c e n t r a t i o n in the a m n i o t i c fluid.
that
to the
the
increase
Foetal lungs and skin have been suggested to release PGF iz the a m n i o t i c f l u i d (19,20). A s i6 g n iQ f i~ c a n t a m o u n t of ~~8) - P G F 2 a a n d its m e t a b o l i t e s w e r e f o u n d in the f o e t a i a d r e n a l ~ s k i n , liver, k i d n e y , a n d m u s c i e . However the foetus has been shown to succ b w thin a few hours of hypertonic saline a inistration (21). In the present stu~y, a s i g n i f i c a n t i n c r e a s e in P G F 2 a l e v e l s was d e t e c t e d a f t e r 8 to i0 h o u r s o f s a l i n e a i n l s t r a t i o n j in c a s e s w h e r e t h e ~i n d u e t ion- ab o r t i on in t e r v a I w a s r t h a n 24 hours. It thus a p p e a r s t h a t the i n c r e a s e in P G F 2 a l e v e l s in t h e n i o t i c f l u i d c o n t i n u e s e v e n a f t e r the d e a t h o the foetus. It j thereforep possible that other sources
also be contributing to the increase i n PGF2a l e v e ! s besides its synthesis in var ous foetal co artments. F u r th e o re, G r e en a i. ( 22 ) h a v e s u g g e s t e d that G F~ a p r e s e n t in the a m n i i l u i d ~ is r e s o r b e d into the f o e ~ a ! c i r c u l a t i o n and ~ e t a b o l i z e d by b o t h the f o e t a l t i s s u e s a n d t h e p l a c e n t a. S i n ¢ e d e c idua I ¢ e 1 ls h a v e b e en s h
DECEMBER
1975
V O L . 12 NO. 6
3
CONTRACE
to
be
ION
rich
in
PGF2~ content (25), it is possible that h y p er t o n i c sa l i n e c a u s e s r e l e a s e o f P G F Z = f r o m t h e s e c e I I s. E x t r a- a i o t i c h y p er t o n i c s a I ine has been sho to cause degeneration of he decidual cells £ i n a l l y r e s u l t l h g in t h e r e i e a s e o f P G F 2 ~ in the a m n i o t i c I u i d ( 1). T h e s e s t u d i e s a l s o p r o v i d e an e x p l a n a t i o n to o u r p r e v l o u s ~ e p p r t s [4) on the d i f f e r e n c e in t h e s e r u m h o r m o n e p a t * e r n s o f p a t i e n t s d u r i n g in d u c t i o n- a b o r t i on i n t e r v a 1 after intra-amniotic administration of hypertonic saline and prostaglandlns. T h e f a i l u r e to d e t e c t a s i g n i f i c a n t f a l i in p i a c e n t a l h o r m o n e s , p r o g e s t e r o n e and h an p l a c e n t a ! i a c t o g e n , l o n g b e f o r e t h e e x p u l s i o n of t h e f o e t u s a n d the s l g n i f l c a n t i n c r e a s e in he c o n c e n t r a t i o n o f P G P 2 s as t h e tlme of abortion approached suggested that hypertonic saline m a y n o t h a v e a d i r e c t .. a..c t i o n on the p l a c e n t a l s t e r o i d o g e n e s i s , mo p r o b a b l y it triggered PGF2~ release in the amniotic f l u 1"d . The present investigations thus l~d to the c o n c l u s i o n t h a t b e s i d e s the t o x ic act i o n o f h y p e r t on ic s a i ine o n f o e tus, it p r o d u c e s a m a r k e d i n c r e a s e .in ... t h e c o n c e n t r a t i o n o £ P G F 2 in t h e a m n i o t i c f l u i d w h i c h m a y be s i g n i f i c a n t l y c o n ribut~Ing to the a b o r t i f a c i e n t p r o p e r t l e s o f 20 p e r c e n t h y p e r t o n ~ c s a i ine.
AC
OWLEDGEMENT
This w o r k w a s s u p p o r t e d b y g r a : n t s f r o m t h e Wo r i d H e a 1 t h Organization. We a r e g r a t e f u l to Dr. Lawrenc.~ L e v i n e ,
Brandels Unlversity, antlserum. b y Dr. J.E. rk was
Massa husetts,
FE
I.
for the supply of PGF2
PGF2~ for radioi unoassay was kindly supplied Pike o~ the Upjo Co. PGF Z~ f o r the c l i n i c a l f r o m K a r o l i n s k a Ins t i t u C e , S t o c k h o l m .
ES
Gustavii, B.: Studies on the mode of action of intra a iotically and eXtra-a iotically :injected hypertonic s a I i ne in t her ap eu t Ic ab o r t i on. A ¢ t a Ob s t e t . Gyn e c o I. Sc d. S pl. 25 1-22, i973. .....
DECEMBER
1975
VOL.
12 NO.
6
CONTRACE
1
B e n g z s s o n , L. a n d C s a p o , A. : O x y t o c i n r e s p o n s e w i t h d r a w a i a n d re in £o r c e m e n t o f de f e n s e m e t h a n i s~ o f the h a n u t e r u s at m ! d p r e g n a n c y . . J. O b s t e t . ~ G y n e c o l . 8 5 : 1 0 8 3 .... 1093, 1962.
Q
Osborn, .H., G . . . . , C.P. a n d Y a one, M.E.'. Response of per plasma progesterone concentration to in t ra-amntot rtonic s a l . . J. Obstet. Gyn oi. I 0 1 : I 0 7 3 - I 0 7 7 , i968.
t
La as, K . R . , R a an, S . A . , H i n g o r a n i , V., P u r i , C . P . , a l l"g a , N. a n d L a as V.: H o r m o n a l c h a n g e s in p a t i e n t s und e r g n g t h e r a p eut i c ab o r t i o n w i t h p r o s t a g I a n d in F~ a, 15 (S) ~ "methyl-pros taglandin P2a and hypertonic sal~ne. C on t r a c ep t £ o n i 0: 517 -6 36, 1974.
4 O
B
P u l k k l n e n , M.O. a n d K i v i k o s k i , A : E f f e c t o f i n c r e a s e d " v " ty d u r l n g n o r m a i uterine vol e on u t e r l" n e actli midtrimester pregnancy. Int. J. G y n e c o l . O b s . 7:172-182, 196 9. G u s t a v i i , B. a n d G r e e n , K.: R e l e a s e of p r o s t a g l a n d i n F2 following inject!on o~ hypertonic saline for therapeutic abortion: A preliminary study. . Obstet. 5 Gynecol. Ii4:1099-ii00, 1972.
Q
J o n e s , Q.L., Clarke, A. and Shutt, D..- A mode of action O f h y p e r t oni c s a l i n e i n i n d u e ing abo r t ion. . J. Ob s t e t. 5 Gyne¢ol "!21 5 5 8 - 5 5 9 , 1975.
0
La as, E.R., Purl, C.P., Hingorani, V. a n d A g a r w a l , N.: Effect o£ hypertonic saline on prostaglandin F2~ levels and dlsappearance of prostaglandln F2~ in t h e a iotic f l u i d of~ m e n ..u..n d e r g o i n g t e r m i n a t i o n o £ p r e g n a n c y w i t h i n t r a ...a. i O t i c h y p ert o n i c s a i i n e I n t e r n a t i o n a I C o n fe r e n c e on P r o s t a g l a n l d Oi n s O M a y ~2 6 - 3 0 , 1 9 7 5 F l o r e n c e P * 200 O
8.
Ca1 ell, B.V., Burstein, S., Brock, W.A. Ra d i o i noas s ? f 5 the F p r o s t a g i d in s Endocrinol. Metab .... 3:17i-1 5, 1971.
.
0
•
1
•
and J.
Speroff, Ci i n.
L. :
Pace-Asciak, Co, life, L.S., Giilett, F . G . a n d 0KXnch, R . A . : Disappearance off pros~aglandin P~a from h an a iotlc £i u i aft e r in t r a- a i o t i c i n3 e c ~ i o n . P r o s Za g i a n d i n s 1 . ' 4 6 9 4 7 7 , 19 Z. S elsson B., en, U., Behrman, H., Granstrom, E., G r e e n i ~ K." a n d J a £ f e , B.M.: Ro d,table ~discussion on analytical methods. Adv ..... Biosci. 9.- 1 2 1 - 1 2 3 , 1973.
DECEMBER
!975
....
L.
12NO,
6
695
CONTRACE
ION
IZ.
L e v l n e , L. glandlns. 1970.
and an, Blochem.
13.
Granstrom,
E. :
Metabolism
S ub j e c
In:
"P r o s t ag land in s
s
V.H.: B±ophys.
tlgenic R e s . Co
of
activlty of prostan. 4 1 : 1 1 7 1 ~ 1 1 7 7 ,
prostaglandin in
S. B e r g s t r o m , K G r e e n a n d B. S a m u e l s s o n , s k a Ins t. S t o c k h. 1 8 - 5 7 , 1 9 7 2 . 14.
15.
o
ferale
c on t ro I
2"
W.H.O.,Karolin-
Brewer, L . L , .... J o h n s o n , W.L. and nter, C.A.: Effects of in t r a- a i o t i c in j e c t i o n o £ h y p e r t o n i c s a i In e o n u t e r i n e a c t i"v" iii t y , b i o o d a n d a m n i ot ic f l u id v o I e s, a n d e i e c t roi concentrations. . 5. O b s t e t . ~ G y n e c o l . I 0 1 : I 0 7 8 1082, 1968. Plentl,
NY. 16
in
Fe r t 11 i t7
A.A. :
Acad.
Sci.
The
dynamics
75"746-761,
of
the
a
iotic
fluid.
Ann.
1959.
Csapo, A.I. : The te ination of pregnancy by the intraa m n i o t i c i n j e c t i o n o f h y p e r t o n i c s a ! i n e. I n: "Y e a rb o o k o f O b s t et. ~ G y n e c o i." J. P. G r e e n h i i I, 1 2 6 - 1 6 ~, 1 9 6 6 .
17.
C s a p o , A ....I. : T h e p r o s p e c t s o f P G s in p o s t c o n c e p t i o n a i t h e r apy. P r o s t a g i a n d ins .3 : 245- 289, 1973.
18.
G r e e n , K., B y g d e m a n , M., T o p p o z a d a , M. T h e r o l e o f p r o s t a g i a n d i n F2C ~ in h u m a n . J.
19.
Obst.
P i p e r , P. from lung
363-38 5,
~ Gynecol.
~ n d V a n e, and other
120:25,31,
J. : T h e tissues.
19
and
Wiqvist,
N. :
4.
r e i e a s e o f p r o s t a g l a n d ins A n n . NY. A c a d Sc£. 180".
1971.
20.
J o n s s o n , C.E. a n d A n g g a r d , E . " B i o s y n t h e s i s and metabolism o£ prostaglandin E in h an skin. S t a n d . J. Clin. Lab. invest. 29:289-296 1972.
21.
K o v a c s , L., R e s c h , B., S z o l l o s ! , J. a n d H e r c z e g , J. : T h e ro I e o f £e t a i de at h in the proc e s s o f therap eut i c abo r t ion induced by intra-a iotic injection of hypertonic saline. J. O b s t e t . G y n a e c o l . Br. C o o . 77: 1 1 3 Z - 1 1 5 6 , i 70.
22.
G r e e n , K., By g d an, M. a n d W i q v i s t , N.: Kinetic and metabolic studies of prostaglandin F2~ a inistered £ntra-a iotically for induction of abortion. L i f e Sci.
14:2285-2297,
23.
1974.
K a r i m , S.H. M. : P h y s i o 1 o g i c a I r o I e o £ p ro s t a g I an d i n s in t h e c o n t ro I o p a r u r i t i o n .... an .. d m e n s t r u a t i on. J. R e p r o d . Fertil. S vl. I 6 ...... .105-119 . 1972.
DECEMBER
1975
VOL.
12 NO.
6
C.Y. P U R l N. B A L I G A , N. V . HIN130 N1 AND K . R .
AGGAR L LA S
De p a r treen t o f R p r o d u c t x4 v e ,, B i o l o g y a n d the o f Ob s te t r ics a n d G y n a e c o i o gy All
IN
Department
I n d i a I n s t i t u t e of M e d i c a l S c i e n c e s New D e l h i - I f 0 0 1 6 , iNDIA
ABST
CT
Int ra- a m n £ o t ic a d m i n i s t ra t i o n o f 2 0 p e rc e n t h y p e r t o n i c s a l i n e c a u s e ~~',~ a s lo g n mif i c a n t increase in t h e c o n c e n t r a t i o n o P G F 2 a in the a m n i o t i c fluid o£ women undergoing second
trimester a b o r t i oQn • The increase ie n PGF2a w a s s l o w i n " .e patients with longer abortion tim Interestingly, x"n o n e case (GS), the levels o f PGF a i n c r e a s e d from 0.32 to 7.48 ng/ml w n the induction-abo tion time was 7.75 hours. H o w e v e r , no s i g n i f i c a n t c h a n g e in P G F z a l e v e l s w a s o b s e r v e d in the p e r i p h e r a l v e n o u s b l o o d d u r i n ~ i ~ d u c t i o n - a b o r t i o n interval. T h e d i s a p p e a r a n c e o f (96--~H)-PGF ~ a n d its rectah o l l i e s w a s s l o w in t h e a ~ n i o t i c f l u i d of s l i n e ad "nist e r e d c a s e s as c o a r e d w i t h the c a s e s w h e r e s a l i n e w a s not g i v e n . T h e s l o w d i s a p p e a r a n c e o f P GF2a a n d its m e t a b o l i t e s in the s a l i n e a d m i n i s t e r e d a m n i o t i c f l u i d h a s b e e n u g g e s t e d to be p a r t l y due to t e r a p i d c h a n g e in a iotic uid volume a~d increased synthesis/release o f P G F 2 a in the a iotic fluid. F r o m the a m n i o t i c f l u i d , P G P 2 a w a s diff u s e d i n t o the f o e t u s w h e r e v a r i a b l e a m o u n Z s w e r e o b s e r v e d in d i f f e t e n t t i s s u e s. The s e o b se rva t io n s sug g e s te d that f o l ! o w i n g the a d m i n i s t r a t i o n of hypertonic saline, an i nc re a s e d e n d o g e n o u s s y n t h e s i s / r e i e a s e o f P GF 2a in he a m n i o t i c f l u i d w a s o b s e r v e d w h i c h m a y a c t o n the u t e r u s l o c a l l y a n d c o n t r i b u t e to the a b o r t i f a c i e n t p r o p e r t i e s o f 20 p e r C e n t h y p e r t on i c s a I i ne.
Accept
e
u for
DECEMBER
pubiication
1975
VOL.
~epte~ber
12 NO.
6
~0,
1975
679
CONTRACE
ION
INTRODUCT ION Since the eariy 1960s intra-uterine administration of hypertonic saline has been widely used for the t e r m i n a t i o n o f m i d t r i m e s t e r p r e g n a n cy. W h e n c a r e f u I Iy and has a a d m i n is t e r e d , t h i s m e t h o d is h i g h l y s u c c e s s f u l A multitude I o w mo r b i d i t y r a t e w i t h f e w s i d e e f f e c t s . fall, foetai death, o f p o s s i b le f a c t o r s l i k e p r o g e s t e r o n e i n c r e a s e d i n t r a - u t e r in e p re s s u r e a n d o x y t o c in- I ike a c t i o n h a v e b e e n p t o p o s e d t o e i u c i d a t e t h e ah o r t i f a c i e n t ac t i o n There are varying reports on the o f h y p e r t o n i c s a l i n e [i). saline on serum hormone levels. A effect of hypertonic few s u g g e s t e d t h a t h y p e r t o n i c s a l i n e l o w e r e d s e r u m p r o ievels (2), w h i l e o t h e r s do n o t f i n d d e p l e t i o n gesterone I n o u r e a r 1 i e r s t u d i e s (4), in p r o ge s t e r o n e I e v e I s [ 3 ) . a significant decline in serum progesterone w e d i d no t f i n d a n d human p i a c e n t a l l a c t o g e n levels after i n t r a - a iotic administration of hypertonic saline, whereas PGF~= and 15(S)15-methyl-PGP2~ produced a significant f a l i in t h e i e v e i s of these hormohes during the induction-abortion interval. Extensive increase in i n t r a - u t e r i n e volume by intra-amniotic administration o f i s o t o n i c s a l i n e f a i l e d to i n d u c e a b o r t i o n (5). G u s t a v i i ( 1 ) s ug g e s t e d t h a t t he ab o r t i f a c i e n t a c t i o n of hypertonic saline was probably d u e to t h e d e g e n e r a t i o n of decidual cells. P r e i i m i n a r y r ep o r t s w i t h e x t r a - a i o t i c h y p e r t on i c saline have sho i n c r e a s e s in P G F 2 ~ l e v e ! s in t h e a m n i o t i c f l u i d (6,7). However, t h e r e is n o s y s t e m a t i c s t u d y to v e r i f y this Inc ease after intra-amniotic administration o f 200 m l o f 20 p e r c e n t h y p e r t o n i c sa!ine. It was with this aim that the disappearance of ( 9 $ - 3 H ) - P G F 2 a , a n d its m e t a b o l i t e s in the amniotic fluid of saline admini stered cases have been c o m p a r e d w i t h t h e o n e s in w h i c h s a l ine w a s n o t g i v e n , The levels of PGF2 in the a iotic fluid and blood have been m e a s u r e d a f t e r .t ...h e a inistration of hypertonic saline. The distribution o f 5 H - P G P 2 ~ in t h e f o e t a l t i s s u e s h a v e a I s o b e e n i n v e s t i g a t e d. A r e i i m in a r y r e p o r t h a s b e en p r e s e n t e d (8). T E R IAL S A N D
METHO D S
Sub ~ects
T w e n t y - t h r e e h e a I t h y w o m e n , b e t w e en 16 - 20 w e e k s g e s t a t ion, s e e k i n g m e d i c a l t e r m i n a t io11 o£ p r e g n a n c y were randomly selected for the present studies, in t h i r t e e n cases, pregnancy was terminated with intra-a iotic PGF2a w h e r e a s t h e o t h e r t e n w e r e g i v e n 200 ml o f 20 p e r c e n t s a i ine int r a-a i o t i c a I ly.
680
CEMBER
1975
VOL.
1 2 N O ....6
CONT 4
In t r a - a
Q
~
I
Q
io t i c a d m i n i s t r a t i o n
of
p
CE
ION
GF 2
With due aseptic precautions, a n n ot was per rmed er abdomen and polyethylene catheter threaded through P h e 1 8 - g a u g e n e e d l e as m e n t i o n e d in d e t a i l e a r l i ~ ~ (~)H" F i r s t d o s e o f 25 m E P G F 2 ~ w i t h 50 to 50 ~ Ci o f ( - ) P GF~was injected throug~ this catheter and t second dose of only radioinert PGF2~ was given after six hours. Q
t
~
I.
.
.
.
3
(gB-SH]-PGF2 630 to 50 :aCi), s p e c i f i c a c t i v i t y 7.5 ~ C i / ~ m o i e, in 5 ~ I s a I ine w a s i~ j e c t e d i n t o t h e a m n i o t i c f l u i d in f o u r w o m e ~ i2 h o u r s p r i o r to t h e i n j e c t i o n of 25 m g P G F 2 a . I n t r a - a n n i o t.... i", . '::..a ......d ..... m.......i ...... n......i ...... s...... t...... r....a.......t ......i ..... o....n.............o.......f...........h ..................e ......r ......t ......o...... n......i.....c ............s......a......I......i.....n ..... e................... ~,o
i
|
200 Millilitre o f 20 p e r c e n t s a l i n e w a s a d m i n i s t e r e d intra-a i o t i c a l l y o v e r a p e r i o d o f a b o u t 20 m i n u t e s in six cases. I ediately following t h a t , 30 to 50 ~ C i o £ (gs- ~ H ) - P G F 2 ~ w a s in e c t e d i n t o t h e a iotic flui through the sa~e catheter.
lntr
a
a
i o t ic
...........................
admin i s t r
-,.
o:f _~h y p e
on "
/ ...................................................... ...........: ...........................................................................................................................
As ~entioned a b o v e , 200 m l o f h y p e r t o n i c in j e c t e d i n t ra- a i o t i c a I 1y i n a n o t h e r S r o u p PGF2~ was not injected in t h i s g r o u p . Co i i e c t ion the
a
o f b Ioo d and
n i o t i¢ f l u i d inistration
a n n io t i c
saline o f fi
was women.
£i u i d
a n d b 1 o o d s a m p I e s w e r e co 1 le c t e d b e fo re of hypertonic s a l i n e or P G F 2 a a n d t h e r e -
after long
at regular i~tervals as free fiow of liquor c a t h e t e r. Ca 1 c u l a t i o n
..............................
o £ ha lf-I
till was
the patient mainta ~ed
abSrted through
or the
so
i £e
To calculate t h e h a l f - ire t i m e o f ( - S H ) ....P G F 2 and its metabolites in t h e a m n i o t i c fluid, the best fit i ines were dra by the me thod o £ two var lab I e I i~ear D e p e n d "i n g u p n t h e p a t t e r n o f d "i s a p e a r a n t e, re g r e s s i o n. single line was dra in cases where linear pattern was observed a n d t w o l i n e s in c a s e s w h e r e b i p h a s i c p a t t e r n fo i 1 o w e d. I ~ e a c h c a s e, t he c o r r e I a t i o n c o e f f i c i e n t w a s greater than 0.95. Pro~ the slope of each line the first ( a t l / 2 ) a n d s e c o n d ( S t l / 2 ) h a l f - i f ' y e s w e re c a l c u l a t e d .
DECEMBER
1975
VOL.
12 N O . 6
681
C
TR
Uvtake
E
+I+ON
o£
and
it s m e t
ol ite s
S Ue S
P o e tus
wa s
co 1i e C te d
a long
w i~ t ~h.
p. I a. c e.n t a
ii
e di a t e ly
after the patient aborted with the use of radioinert and 5H-PGF2a t It w a s t h o r o u g washed with chilled saline and d isse c te d ou t to co i Iec t a I E i s s u e s, A d r e n a i, T a in j k i d ~ e y , i i v e r, I u n g s, m u s c le, s k n p i acen t a and u~b i i i c a 1 cord were minced, washed, dissolve~ in s o i u e n e a n d c o u n t e d in t o l u e n e b a s e d s c i n t i l l a t o r . ~n t w o c a s e s , f o e t a l t i s s u e s re e x t r a c t e d w i t h ~ e t h a n o l : c h l o r o f o r m and the dried e x t r ac t
c o u n t e d. f _ .......r a
ioact
T r it i con t ent s m e a s u r e d in t h e a i i q u o t s o £ s e r and a iotic fluid s i t s in d u p l i c a t e u s i n g t o l u e n e ...b .ased scintillator with biosolv ( B e c k m a n ) as w a t e r e m u l s i f l e r ~n a N u c l e a r C h i c a g o l i q u i d sc i n t i l l a t z"o n c o te r Mark If. Remaining s les w e r e ~ r o z e n in a l i q u o t s b e l o w - 1 5 ° C . Ex r
ct..ion........a......n.....d
chroma
gr
h y ....
P G F 2 a w a s e x t r a c t e d f r o m a ~ n £ o t ic £ 1 u i d b y t h e m e tho d o C a I d w e i I e t a 1. (9). In b r iv f, n i o t ic f i ui d w a s r e n d e r e d acldlc to 4 . 0 ) w i t h O . 1 N H C I a n d e x t r a c t e d w i t h
diethxl
The
chromatographed
e~tract
was
washed
with
water,
dried
and
through
s£1ic£c acxd col ns u s i n g i n c r e a s i n g ~ethanol in b e n z e n e :e t h y l a c e t a.t.e. . . . ( 3; 2 -) .
concentrations of Eluted PGF2a was dried
and
f urther
subjected
t h in
to
1
er
chromatography in c h l o r o f o r m : t h a n o l :a c e t i c a c l d : w a t e r (90:9 : I:0.6) system (I0). PGF2a zone parallel to t h a t o f a u t h e n t i c P G F 2 a w a s e x t r a c t e d a n d co ted £or he t r i t i conte~t. Ra~ £o
no
s
PGF 2 a
0.2 to 12-ng PGF~. To r u n t h e a s s a y , about ~ 500 cp~ of 3H-PGF,~ was added to eacfi sample and then ~extracted and chro~atograPhed ~ through sil£cic acid colu s (9). The ¢luant was dried a~d made to one ml With ethanol. An a l i q u o t was taken each sa to account f o ~ ... r e c o v e r y , which fro~.65 t o 80 percent. ~he remaining s le s dried a n d ~ a d e t o O. I m l with 0.01 M Tris-HCl buffer (pH 7.4) containing 0.15 M NaCl and 0.! p e r c e n t g e l a t i n . 0.I MI diluted 1 / 4 0 0 g a v e a b o u t 55 p e r c e n t .binding)
tubes. ature
682
The for
t
es
were
30 m i n u t e s .
ently ~H_PGF vo rtexed 2~
(8000
antgser was added
and cpm)
~DEcEMBER
left
(a d i l ~ t i o n o f to all t h e a s s a y
at
-in 0.i
1975
r o o m ~te ml
buffer
VOL.
12
was
. 6
C
T R
E
I
a d d e d to all the tubes and the r e a c t i o n m i x t u r e i" n c u b~,ated at 37"C for one hour. This w a s ! l o w e d b y the a d d i t i o n o £ 0 . 1 ml o£ 1 / 1 0 0 n o r m a l r a b b l t s e r u m a~d t h e n 0.I ml o£ g o a t a n t l s e ~ to r a b b i t ga a globulin. The 01e r e a c t i o n ~ixture was left ove rnight
a~
Z to
4 "C
and
c e nt r i fuged
at
55 O D rp~
£or
40
m £ n u t es.
The pre~ :ipitate w a s d i s s o l v e d :in 0 2 ml o£ 0 I N N a O H and c o u n t e d c. To t the s e n s i t i v l t y a"n d r r o .c i b i l i t y of the a s s a y m e t h o d , d i f f e r e n t c o n c e n t r a t i o n s of r a d l o i n e ~ t PGP2a w e r e a d d e d to 3 mi of s e r u m oT ioric f l u i d and the s les extrac~ed, ¢ h T o m a t o g r a p h e d and a s s a y e d as d e s c r i b e d above. Nonspeciflc b i n d l n g was a b o u t ~ to 2 p e r c e n t en the a n t l s e r was~rep!aced w i t h n o r m a l tab it s e r u m ..... The l e a s t d e t e c t ~ b l e level o f P O P 2 a for l l n e a r l z e d curve, f r o m 0.6 to 1 0 ng~ was 0.8 ng.
RE SULTS earance
~i
of PGF2~ --~::_-
from a
iotic ..........
;::_n
fluid ~-w
~
T w e n t y flve ~ i l l i g r a m s o£ P G F 2 ~ w i t h ( 9 ~ - 3 H ) - P G F z s was a £nistered to five wo~en, o t h e r dose of r a d i o i n e r t 2S mg PCF2~ w a s g i v e n a f t e r six hours. The a m n l o t i c ffluid s a m p l e s were e x t r a c t e d and c h r o m a t o g T a p h e d to a n a l y s e PGP2a c o n t e n t at v a r i o u s t i~tervals. As sho in Fig. i, the r a t e of d i s a p p e a r a n c e of PGF was d i f f e r e n t in d i f f e r e n t cases. In the five cases- AP ~P, C P , D P , E P - 62.5, 38.6, S0.0, 45.8 and 43.8 percent, r e s p e c t i v e l y , P GFz~ d i s a p p e a r e d in 24 hours of P G F 2 a a inistratio~. On an averages 45 8 p e r c e n t PGP2a d i s a p p e a r e d in 24 h o u r s r near the tl of abortion. Dis
p
rance..Of 5 H T P G F 2 ~
and
its m e t a b o l i t e s
from
The dis p e a r a ~ c e o £ 3 H - P G F 2 ~ a~d its m e t a b o l i t e s in the a m n i o t i c £ 1 u l d of r wo~en a i n i s t e r e d w i t h N0 to S0 ~Ci o£ 5 H - P G P 2 ~ is sh n in Fig. 2. In a l l ~he ff r cases, r a p i d d e c l i n e of H - P G F 2 a and its m e t a b o l i t e s was o b s e r v e d to six hour s o£ 3H-p GP 2 a i n i s t r a t ion. T h e a v e r a g e f i r s t hal f- li fe b e i n g 5.3 + 0.7 (me an ÷ S.E.) h o u r s . 3.5 to 6.7 hours). w e v e r , £W two c a s e s , ere a iot£ d sa los c o u l d be c o l l e c t e d up to 12 h o u r s of P G F 2 a a d m l n i s t r a t i o n , he d i s a p p e a r ante off 5 H - P G F 2 ~ and its metaboiites s l o w e d do to a s e c o n d halff-life of 9 ÷ 0.6 hours.
CEMBER
1975
V O L . 12 NO. 6
CONTRACE
ION
A
~k
T
gO"
60. ~CP
,AP
i
G.
.
.
.
.
.
4
.
W:
................................
S
Figure
1.
Dis flu
...........
~ .........
! 0
U
A
S
earance of prostaglandin F from amniotic of wo~en after i n t r a - a m n i o ~ c injection.
(gs-3H)-PGF~a was injected intra-a~niotically along with 5 ~ g P G F 2 ~ in f i v e women. Another 2 5 mg d o s e o ~ PGF2m w a s g i v e n after 6 hours. niotic fluid samples were extracted and chromatographed to deter ine the amount o~ PGF2~ d i s a pp e a r a n c e a t v a r i o u s t i~ e i n t e r v a 1 s.
684
DECEMBER
1975
V O L . 12 N O . 6
NTRACE
laO00
ION
-
l0
8 000-
0
6000-
_u
O
_o
E
0
:~ e ooou
0
2
4
6
8
i0
HOURS
Figure
2.
D i s a p p e a r a n c e of C g B - 3 H ) - p r o s t a g l a n d l n F2a and its m e t a b o l i t e s from nlotlc fluid o£ w~men a ft e r
int ra- a
i ot ic
inj e c t i on.
4
( 9- 3H) -PGF2} wa s ~nj ec t d tnt ra- a i o t .ic a 1 1 ) , in a gro o four women, iotic fluid s les were ana lys ed £o PG .F 2 a a n d i t s m e t a b o i i t e s c o n t e n t a t va r i o u s t ime i n t e rv a I s. Ra d i o i n e r t PGF s not g i"v e n t o t h i s g r o u p of cases
dur
CEMBER
1975
g the course of our study.
VOL.
12 N O .
6
CONTRACE
ION
Dis
O~
e
ante
3H,
2a a
_itS_
_ta_bol
s from
amniotic
T h e dl s a p p e a r a n c e o 3H. P G F ~a. and i t s m e t a b o I i t e s f r o m t h e a m n i o t ic ffluid o f w o m e n a d m in I s t er ed 2 0 0 ml h y p e r t o n i c s a l i n e is s h o in Fig. On t h e s e m i ! o g s c a l e , in 4 of the 6 c a s e s j the d l s a p p e a r a n c e s h o w e d a b i p h a s i c p a t t e r n . The i n i t i a l d l s a p p e a r a n c e o f 3 H . P G F 2 a a n d its m e t a b o l l t e s w a s fast, an a v e r a g e f i r s t h a l f - l i f e b e i n g i 0 . 0 + 2.3 h o u r s (range 3 . 3 to 18.8 h o u r s ) . However, afterwa s, the d i s a p p e a r a n c e was s l o w e d do to an a v e r a g e s e c o n d h a l f ...l. i f e o f 24.2 ÷ 5.8 hours. In two c a s e s (LS, KS), l i n e a r dis t r l b u t ion p a t t e r n w s o b s e r v e d w i t h a h a l f - l i f e o £ IZ.0 a n d 18.8 hours, respectlvelv. and A co a r i s o n of the f i r s t a n d s e c o n d h a l f - l i f e of P G F 2 a its m e t a b o l i t e s in the a m n i o t i c f l u i d of n o r m a l a n d h y p e r t o n i c s a l i n e a d m i n i s t e r e d c a s e s is p r e s e n t e d in T a b l e I . I t c a n be s e e n t h a t the f i r s t h a l f - l i f e w a s i n c r e a s e d f r o m 5.3 + 0.7 h o u r s to 10.0 ÷ 2.3 h o u r s . H o w e v e r , this d i f f e r e n c e was n t s t a t i s t i c a l l y significant. On the o t h e r hand, a s i g n i f i c a n t i n c r e a s e ~p~0.01) in the s e c o n d h a l f - l i f e o f P G P 2 s a n d its metabo.iites w a s o b s e r v e d in c a s e s a d m i n i s t e r e d with hypertonic sallne, in b r i e f , a p r o l o n g a t i o n of the h a l f - l i f e t i m e of P G F 2 ~ a n d its metabolites w a s o b s e r v e d in c a s e s i n j e c t e d w i t h hypertonic s a l i n e PGF2~
levels
in a m n i o t i c
fluid
The levels of PGF2a were measured by a radioi noassay ich h a d a s e n s i t i v i t y o f 0.8 ng. The r e l i a b i l i t y of radioimmunoassays as a t e c h n i q u e ffor the m e a s u r e m e n t of PGP2 le-~els has y e t to be e s t a b l i s h e d (I!). H o w e v e r , the a n t i b o d i e s u s e d in the p r e s e n t s t u d i e s b i n d m o s t e f f e c t i v e l y w i t h the h o m o l o g o u s a n t i g e n - P G P z a . A b o u t 33 n g of P G F 1 330 n g of 1 5 - k e t o - P G P 2 a and m o r e t h a n 1 pg o f 1 5 , 1 4 - d i b ~ d r o ! S - k e t o - P G P 2 a w e r e r e q u i r e d for 50 p e r c e n t i n h i b i t i o n v a l u e s
(12).
The effect of hypertonic saline on PGF2~ levels in the a i o t i ¢ f l u i d is sho in Fig. 4. The l e v e l s of P G F 2 a s t a r t e d i n c r e a s i n g a f t e r the a inistration of s a l i n e . In t h r e e c a s e s (HS, IS, JS), w i t h an a v e r a g e i n d u c t i o n - a b o r t i o n time of 2 8 . 7 5 h o u r s , i n c r e a s e in P G F 2 s l e v e l s s t a r t e d a f t e r 8 - I 0 h o u r s o f saline administration, a n d n e a r the t i m e of a b o r t i o n , a s i g n i f i c a n t i n c r e a s e in P G P ~ a l e v e l s h a d o c c u r r e d In one c a s e (GS), w h e r e p r e g n a n c y e r m i n a t e d aft 7.75 h o u r s of saline a i n i s t r a t i o n , a s h a r p i n c r e a s e in P G F z a l e v e l s w a s observed. I n this case, P G P 2 a l e v e l s i n c r e a s e ~ f r o m 0.32 t o 7.48 n g / m l a m n i o t i c fluid.
1
DECEMBER
1975
L. 12 N O . 6
I0
COS) ~ t
=6"3HR t
A
~N ~ME
HR
i ;i
OS 5 J
6
.
.
.
.
.
.
ii!~i~
30.1
g 4
MS
Z
~S 0
J5
4
2
6
8
i0
12
!4
16
18
20
22
26
HOURS
~£guTe
3.
Dis i t s
pearance of f9g-3H)-prostaglandin ~ e t a b .o i i t e s £ r o ~ hyp e r t on I c s a I I ne
tered
a
iotlc
fluid
of women
F a
and !ni s -
undergoi:ng abortlon.
3 ...... p C9~-H ) GF2 was injected i d l a t e i....y a f t e r t h e a inlstrat[on o 200 m l off 20 p e r c e n t s a l l n e in s Ix wo~en • i o t Ic f l u i s [ e s wet e iys ed to d e t e £ n e t h e d l s a p p e a r a n c e o £ P G F 2 a a n d Its me t ab o i i t e s a t v a t i o u s t e int e a is 2 Rad io I n e r t P G F 2 a w a s n o t g l v e n to t h e s e c a s e s .
DECEMBER
1975
VOL.
12 NO.
6
687
b~
)
~a
b
N.S.
[0.0 + 2.3 (6)
~.3 + 0.7 (4)@
p¢O.O1
24.2 + 5.8 (3) #
9.7 + 0.6 (2)
HOURS
(MEAN + s.E)
(ME.~ + S.E) HOURS
#
In two cases [LS and KS), linear distribution of (gs-bH)-PGF2a and its metabolltes wis observed with a half-hfe of 12.0 and 18.8 hours, whereas in case (OS) sam phng ' was not possible after 6 hours.
@ The figure in parentheses is the number of observations.
** Disappearance of [98-3H)-PGF2a and its metabolites from amniotic fluid of women administered with intra-amniotic hy~ertonic saline.
* Disappearance of {98-3H)-PGF2a and its metabolites from amniotic fluid of cases where saline was not glve " n.
Level of
~ypertonic saline** tdministered cases
~orma] cases*
~ASES
SECOND HALF-LIFE
;!RST H_ALF-LIFE
EFFECT OF HYPERTONIC SALINE ON DALF-LIFEOF (! -PROSTAGLANDIN F2a AND ITS METABOLITES IN .~h?;IOTIC FLUID
tABLE I
Z
m
,q
>
3Z
CONTRACE
ION
GS A
:Y
:J~E
N$
4
8
i2
~6 H
Figure
t
0
u
R
s
Prostaglandin
F2a levels in amniotic fluid o f ,,,omen a d m i n i s t e r e d w i t h £ntra a m n i o t i c h y p e r tonic s a l i n e for t e r m i n a t i o n of p r e g n a n c y .
P G F Z a i e v e I s w e t e me a s u r e d b y r a d i o i noa s say. Ea c ~ s p 1 e wa s e x t r a ¢ t e d a ¢ d c h r o ~ a t o g r a p h e d through silicic acid columns and assayed using a n t i s e r u m r a i s e d a g a i n s t PGF coup led to po Iy£-lysine in rabbits and (9B)-PGF2a (about 7 pmole) as tracero The antibodies used in this study bind st effectively 'ath the homologous a n t i g e n - P GF 2 a -
DECEMBER
1975
VOL.
12 NO. 6
CONTR
EPTION
Is i : : b o d
2
The effect of h ertonic saline on serum levels of PGF2a in five w o m e n u n d e r g o l n g t e r m l n a t i o n of p r e g n a n c y is s h o w n in T a b l e II. No c l e a r p a t t e r n of s e r u m l e v e l s of p G F 2 , c o u l d be o b s e r v e d , in one c a s e (GS), the s e r u m levels o£ P G ~ a be for (l. Z0 n g / m l ) and a £ t e r six h o u r s of s a l l n e a d m i n l s tration (0.95 ng/ml) were more than observed in other cases. The £ n d u c t i o n - a b o r t l n - l n t e r v a l , 7.75 hours, was the l o w e s t in this case. D:istribution
of
3H
2a
and
its
:metaboIites
The di'stribution of 3 H - P G P 2 and its m e t a b o l i t e s in the various foetal compartments, a~ter intra-amniotic a inistration of a s i n g l e dose o£ 40 m g F2 , is s h o i"n T a b l e III. PGF 2~ a n d its m e t a b o l i t e s w e r e f o u a i s t r i b u t e d in d i £ f e r e n t a m o u n t s in a11 the t i s s u e s i n v e s t i g a t e d . The u p t a k e was maxl m in the u m b i i i c a l cord. S k i n and m u s c l e also h a d m o r e radioactivi .... t h a n o t h e r tissues. In three of the £our cases, the u p t a k e ot~ 5 H . P G F 2 a a n d . i t s m e t a b o l i t e s was m o r e in a d r e n a l th in l i v e . T h e s e p r e l l m i n a r y s t u d i e s also s h o w e d a t r e n d which indicated that there was a somewhat higher take of PGP a a n d i t s m e t a b o l i t e s when the foetus was exposed to PGF a for a l o n g e r p e r i o d of time, as s h o w n in case 4.
DISCUSSION The p r e s e n t i n v e s t i g a t i o n s on the m o d e of a c t i o n of i n t r a - a m R i o t i c h y p e r t o n i c s a l i n e for the t e r m i n a t i o n of s e c o n d t r i m e s t e r p r e g n a n c y s u g g e s t e d that 20 p e r c e n t saline gradually increased the levels of PGF2a in the amn £ o t flu i d a s t he t ime o £ ab o r t ion a p p roa ch e d. The d e l a y in he e l i m i n a t l o n o f P G F 2 a a n d i metabolites f r o m the ...a..m n i o t l c f l u i d o f w o m e n a i n i s t e r e d w i t h hypert o n i c s a l i n e also i n d l c a t d the p o s s i b i l l t y of i n c r e a s e d of P G F 2 ~ in t h e a iotlc fluid. observations along with t e e s u l t s ....that ...no . s gnificant c h a n g e in p e r i p h e r a l s e t l e v e l s o£ P G P 2 a w a s . o b s e r v e d d u r i n g the i n d u c t i o n - a b o r t l o n interval s u g g e s t e d that r e l e a s e d i n the a iotic~fluid y on ..the .. Ut
rU
locally
pr p e r t i e s
of
and
contribute
20 p e r c e n t
to
the
ab0rtifacient
saiine.
The h a l f life of PGF2a in longer than in blood. ...... A b o u was l e f t in c i r c u l a t i o n
the a iotic fluid is much 3 percent ....o injected PGF2 o n e and a h a l f m i n u t e o f f
DECEMBER
1975
L .... 1 2 N O .
6
""~'~
7.75 34.50 25.25 28.50
IS
~IS
IS
;S
0.32
0,65
0.48
1.20
0 Hr
0t.27
0.61
0.24
0,95
6 Hr
0.41
0.75
0.55
-
12 Hr
0.51
0,89
0.90
18 Hr
0.46
0.78
0.71
24 Hr
PROSTAGLANDIN F2a LEVELS ....................... ng/ml ........................... _
IN BLOOD OF W O R N ADMINISTE~D WITH INT~-A_~NIOTIC SALINE FOR TERMINATION OF PREGNANCY
PGF l o"els were measured by radioimmunoassay, The v a l u e s have been exp as ng/ml s e r ~ a f t e r necessary c o r r e c t i o n for losses incurred during e x t r a c t i o n and chromatograph)'.
ABORTIONINTERVAL Hr
CASE
INDUCTION-
PROSTAGLb.NDIN F, 1
rABLE II
,q
.4
3,720* " 20,000 ~,I00 1,580 2,580 2,900 4,620 8,410 ~
ADRENAL
PLACENTA
LIMBILICA] CORD
SKIN
BRAIN
I@JSCLE
LUNGS
KIDNEY
LIVER
4,090
4,900
2,520
6,220
-
5,190
16,020
930
5,600
FOETUS 2 (IAT 19.5 Hr)
5,220
6,550
5,400
5,160
1,450
8,520
"
1,160
7,290
FOETUS 3 (IAT 31.5 Hr)
11,700
4,500
19,020
1,650
18,870
23,060
3,120
12,390
FOETUS 4 (I~:T 34.0 Hr)
TISSUES AFTER !NTRA-~IOTIC )STAGLANDIN F2=
*~. . . . . . . ~ are expressed as d i s i n t e g r a t i o n s per minute per gm t i s s u e . (! ~ (40 ~Ci) was administered i n f o u r women undergoing t e r m i n a t i o n of pregnancy w l t h i n t r a - a ~ i o t i c "~ "40 mg). Foetus and p l a c e n t a were collected inediatel7 after the aborted. Foetal t i"s s u e S were washed, minced and counted for the t r i t i u m content.
FOETUS I (IAT 13.0 Hr)
TISSUES
JPTAKE OF P~OSTAGLkNDIN F2o AND ITS METABOLITES BY ' ~MINIST~TION OF RADIOINERT AND (gB-
tABLE I I I
Z
),
Z
CONTR
E
ION
its administration (15). The significant increase (p~O.O1) in the second half-life o f PGP2~ a n d i t s m e t a b o l i t e s in s a l i n e a d m i n i s t e r e d a m n i o t i c f l u i d as c o m p a r e d w i t h c a s e s in W h x~ c h s a l i n e w a s n o t given, c o u l d at l e a s t be due to c h a n g e in a m n i o t l c f l u i d v o l u m e a f t e r the a d m i n i s t r a t i o n o f 20 p e r c e n t s a l i n e a n d i n c r e a s e d e n d o g e n o u s s y n t h e s i s / r e l e a s e of P G P 2 s in the a m n i o t i c f l u i d . S i n c e the a d m i n i s t r a t i o n o f 40 g~ s o d i u ~ c h l o ide p r o d u c e s i o n i c i m b a ! a n c e in a i o t i c fluld, the b o d y f l u l d s e n t e r the a iotic 1me . c a v i ty c au s i ng a c h . a n.g e. in . . a m.n i.o t.l c . £.1 u.l d . v.o l u . m e. w i t h t" It
is
possible
that
the
change
in
volume
af
cted
the
ellmin ion p a t t e r n of P G F 2 ~ and its m e t a b o l i t e s . Brewer et al. ( i ) h a v e r e p o r t e d ~a net i n c r e a s e in niotic fluid 1 assoclated with a decreased blood volume within 2 to 4 h o u r s f o l l o w i n g i n t r a - a m n i o t i c i n e c t i o n of h y p e r t o n l c sallne. Subseque 1y, the a m n i o t i ¢ fl~ id v o l u m e s a r t e d d e c l i n i n g and W i t h i n "8 ~to I0 h o u r s , r e t u r n e d to n o r m a l levels. It has a l s o b e e n r e p o r t e d t h a t the h a l £ - l i f e o£ d e u t e r ium o x i d e i s I e s s in o I igo - h y d r a i o s t h a n in p o lyh y d r a m n i o s ~159. It thus a p p e a r s t h a t in a u d i t i o n other factors, endogenous synthesis/release Of PGF 2 and c h a n g e in the v o l u ~ e o f a m n i o t i c .fluid m a y c o n t r i b u t e to t h e s l o w d i s a p p e a r a n c e of P G F 2 ~ a n d its m e t a b o l i es in the a m n i o t i c f l u i d o£ s a l i n e a i n i s t e r e d cases. F u r t h e r m o r e , C s a p o (16) s u g g e s t e d that the i n c r e a s e in niotic f iuid volume after hy.pertonic s a l i n e t r e a t m e n t f a c i l i t a t e s a b o r t i o n b y e n h a n c i n g uterine v S i n c e it is k n o ~ that p r o s t a g l a n d i n s a r e r e I e a s e d i n r e s p o n s e t o u t e r i n e s t r e t c h , c e 1 i u i a r dama and
to
vaginal
enhanced in P G P 2 s
distention
(17,18),
i t ~is p o s s i b l e
uterine activity may also contribute c o n c e n t r a t i o n in the a m n i o t i c fluid.
that
to the
the
increase
Foetal lungs and skin have been suggested to release PGF iz the a m n i o t i c f l u i d (19,20). A s i6 g n iQ f i~ c a n t a m o u n t of ~~8) - P G F 2 a a n d its m e t a b o l i t e s w e r e f o u n d in the f o e t a i a d r e n a l ~ s k i n , liver, k i d n e y , a n d m u s c i e . However the foetus has been shown to succ b w thin a few hours of hypertonic saline a inistration (21). In the present stu~y, a s i g n i f i c a n t i n c r e a s e in P G F 2 a l e v e l s was d e t e c t e d a f t e r 8 to i0 h o u r s o f s a l i n e a i n l s t r a t i o n j in c a s e s w h e r e t h e ~i n d u e t ion- ab o r t i on in t e r v a I w a s r t h a n 24 hours. It thus a p p e a r s t h a t the i n c r e a s e in P G F 2 a l e v e l s in t h e n i o t i c f l u i d c o n t i n u e s e v e n a f t e r the d e a t h o the foetus. It j thereforep possible that other sources
also be contributing to the increase i n PGF2a l e v e ! s besides its synthesis in var ous foetal co artments. F u r th e o re, G r e en a i. ( 22 ) h a v e s u g g e s t e d that G F~ a p r e s e n t in the a m n i i l u i d ~ is r e s o r b e d into the f o e ~ a ! c i r c u l a t i o n and ~ e t a b o l i z e d by b o t h the f o e t a l t i s s u e s a n d t h e p l a c e n t a. S i n ¢ e d e c idua I ¢ e 1 ls h a v e b e en s h
DECEMBER
1975
V O L . 12 NO. 6
3
CONTRACE
to
be
ION
rich
in
PGF2~ content (25), it is possible that h y p er t o n i c sa l i n e c a u s e s r e l e a s e o f P G F Z = f r o m t h e s e c e I I s. E x t r a- a i o t i c h y p er t o n i c s a I ine has been sho to cause degeneration of he decidual cells £ i n a l l y r e s u l t l h g in t h e r e i e a s e o f P G F 2 ~ in the a m n i o t i c I u i d ( 1). T h e s e s t u d i e s a l s o p r o v i d e an e x p l a n a t i o n to o u r p r e v l o u s ~ e p p r t s [4) on the d i f f e r e n c e in t h e s e r u m h o r m o n e p a t * e r n s o f p a t i e n t s d u r i n g in d u c t i o n- a b o r t i on i n t e r v a 1 after intra-amniotic administration of hypertonic saline and prostaglandlns. T h e f a i l u r e to d e t e c t a s i g n i f i c a n t f a l i in p i a c e n t a l h o r m o n e s , p r o g e s t e r o n e and h an p l a c e n t a ! i a c t o g e n , l o n g b e f o r e t h e e x p u l s i o n of t h e f o e t u s a n d the s l g n i f l c a n t i n c r e a s e in he c o n c e n t r a t i o n o f P G P 2 s as t h e tlme of abortion approached suggested that hypertonic saline m a y n o t h a v e a d i r e c t .. a..c t i o n on the p l a c e n t a l s t e r o i d o g e n e s i s , mo p r o b a b l y it triggered PGF2~ release in the amniotic f l u 1"d . The present investigations thus l~d to the c o n c l u s i o n t h a t b e s i d e s the t o x ic act i o n o f h y p e r t on ic s a i ine o n f o e tus, it p r o d u c e s a m a r k e d i n c r e a s e .in ... t h e c o n c e n t r a t i o n o £ P G F 2 in t h e a m n i o t i c f l u i d w h i c h m a y be s i g n i f i c a n t l y c o n ribut~Ing to the a b o r t i f a c i e n t p r o p e r t l e s o f 20 p e r c e n t h y p e r t o n ~ c s a i ine.
AC
OWLEDGEMENT
This w o r k w a s s u p p o r t e d b y g r a : n t s f r o m t h e Wo r i d H e a 1 t h Organization. We a r e g r a t e f u l to Dr. Lawrenc.~ L e v i n e ,
Brandels Unlversity, antlserum. b y Dr. J.E. rk was
Massa husetts,
FE
I.
for the supply of PGF2
PGF2~ for radioi unoassay was kindly supplied Pike o~ the Upjo Co. PGF Z~ f o r the c l i n i c a l f r o m K a r o l i n s k a Ins t i t u C e , S t o c k h o l m .
ES
Gustavii, B.: Studies on the mode of action of intra a iotically and eXtra-a iotically :injected hypertonic s a I i ne in t her ap eu t Ic ab o r t i on. A ¢ t a Ob s t e t . Gyn e c o I. Sc d. S pl. 25 1-22, i973. .....
DECEMBER
1975
VOL.
12 NO.
6
CONTRACE
1
B e n g z s s o n , L. a n d C s a p o , A. : O x y t o c i n r e s p o n s e w i t h d r a w a i a n d re in £o r c e m e n t o f de f e n s e m e t h a n i s~ o f the h a n u t e r u s at m ! d p r e g n a n c y . . J. O b s t e t . ~ G y n e c o l . 8 5 : 1 0 8 3 .... 1093, 1962.
Q
Osborn, .H., G . . . . , C.P. a n d Y a one, M.E.'. Response of per plasma progesterone concentration to in t ra-amntot rtonic s a l . . J. Obstet. Gyn oi. I 0 1 : I 0 7 3 - I 0 7 7 , i968.
t
La as, K . R . , R a an, S . A . , H i n g o r a n i , V., P u r i , C . P . , a l l"g a , N. a n d L a as V.: H o r m o n a l c h a n g e s in p a t i e n t s und e r g n g t h e r a p eut i c ab o r t i o n w i t h p r o s t a g I a n d in F~ a, 15 (S) ~ "methyl-pros taglandin P2a and hypertonic sal~ne. C on t r a c ep t £ o n i 0: 517 -6 36, 1974.
4 O
B
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