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Study of the effect of uremic metabolites on erythrocyte glycolysis
Study of the Effect of Uremic Metabolites on Erythrocyte Glycolysis By
Attempts
have
been
JEAN
made
effect of uremic metabolites rocyte glycolysis. In blood persons, glucose filtrate
E
the rate is normal. of uremic
XTENSIVE
M. MORGAS AND ROBERTE. MORGAN to study
the
upon erythfrom uremic
of disappearance of However, the ultrablood
produces
inhibi-
tion
of
glucose
erythrocytes. in
uremic
The blood
utilization failure may
by
normal
to observe be
due
to
this the
younger erythrocyte population. The mechanism and possible significance of this inhibition
remain
to be shown.
STUDIES ha\xs ken reported in recent vears concerning essential metabolic processes in the erythrocvte. Erythrocvte metabolism is entirely anaerobic.] The energy derived from glycolysis and stored ;LS the “pump” which exadenosine triphosphate (ATP) lr, ‘: necessary to maintain trudes sodium and water from the cell and retains the necessary high potasGllrn concentration.“,” The maintenance of the cell membrane is also energ\’ rccpiring4 Recent studies of certain congenital hemolvtic anemias ha\rct illustrated the ervthrocvte fragilitv which occurs as a result of glucose-fiphosphate dehvdrogenase; and pyruvate kinasc” deficiency, both leading to impaired glycolvsis. The anemia of severe rlremia has been shown to 1)(x hemolvtic in na&e,’ which suggests the possibility of a metabolic defect. The present study was therefore designed to determine: (1) the rate at whiclI glucose is utilized by erythrocytes from uremic persons under varied conditions: and (2) the effect of ultrafiltrate of blood from llremic persons on normal red blood c,ell metabolism.
630
J.M.
MORGANAND R.E. MORGAN
after 2 hours of incubation. Each incubation was carried out in duplicate. The rate of disappearance of glucose was assumed to be the erythrocyte utilization rate. To prepare the ultrafiltrates, sera from normal and uremic persons was collected and frozen until processed. Each serum was placed in a boiled cellophane membrane and centrifuged in the Tori-Bara apparatus at 2700 RPM for 16-24 hours at 5” C. The ultrafiltrate was kept frozen until utilized for study. At that time 100 mg. per cent glucose was added
and the ultrafiltrate
rected
to 40 per cent and incubations
was mixed
with
fresh
were
carried
erythrocytes.
The
hematocrit
was
cor-
out as indicated.
RESULTS
The glucose utilized by erythrocytes from 10 normal subjects, determined in native plasma, was measured on several occasions. The average for the group of 17 duplicate determinations was 9.6 mg. per cent/hour, with limits of 618 mg. per cent/hour. The data are summarized in table 1. The final pH determinations \,aried from 7.44 to 7.85. The initial blood sugar concentrations were between 34 to 102 mg. per cent with an average of 65 mg. per cent. The rate of disappearance of glucose in blood from azotemic persons was not significantly different from that in the normal group. The average for this group of 9 persons, several studied on more than one occasion, was 10.3 mg. per cent. Initial blood-sugar concentrations averaged 113 mg. per cent, with a range of 34 to 148 mg. per cent. The data are summarized in table 2. Three of the patients studied suflered from acute renal failure and 6 had longstanding moderate to severe uremia. The blood urea nitrogen (BUN) concentrations varied from 39 to 237 mg. per cent, and all but 1 patient had some degree of anemia. No correlation was apparent between either the BUN 01 creatinlne concentration and the rate of glucose utilization. When erythrocytes from healthy donors were incubated in ultrafiltratc from the same or other healthy persons, the rate of glucose utilization was not significantly different from that of the erythrocytes in their native plasma (specified as “control”). The rate of utilization averaged 104 per cent of the control, with all initial blood sugars between 135 and 144 mg. per cent and the final pH’s varying from 7.48-7.84. The ultrafiltrate from azotemic persons was distinctly inhibitory by comparison, with the rate of glucose disappearance averaging only 55 per cent of the control. The data are summarized in table 3. The initial blood sugars varied from 139 to 185 mg. per cent and the final pH from 7.4 to 7.66 mg. per cent. Three of the ultrafiltrates were obtained from patients shown in table 2 (O.T., P.S. and B.C.). Five others were from selrere chronic uremics and one was from an acute renal failure patient just prior to hemodialysis. Clinically, the patients in this group were more symptomatic as a result of renal failure than most of the patients summarized in table 2. An attempt was made to incubate the erythrocytes from uremic persons in normal ultrafiltrate. However, the results were highly variable. In 4 trials, the rate of disappearance of glucose was 36, 107, 121 and 283 per cent of that in the native plasma. Marked discoloration of the ultrafiltrate was noted after
centrifllgation
the cause
of the
of the
first
low utilization.
sample.
This
Since
severely
suggested uremic
that blood
hemolysis may
have
was an
LlRE:JfI(:
METABOLITES
6.31
C:LU(:OLYSlS
Table l.-Glucose Disappearance Rate Blood-Leukocytes Excluded-Hemutocrit 40 Per Cenf
Normal H. Y.
ON EHTTHRO~:\iTE
PH’
*H’
(1)
7.50 7.60
7.31 756
15 .5 1Z!..li
(2)
7.48
7..50 7.65
8.0 x.0
7.53 I31
G.U.
10.0
7.49 7.48
7.48
10.0
11. H.
i I)
7.34 753
7..54 7..55
7.0 7.r,
J.\\.
il)
7.*57 7.57
7.62
12.0 12.0
I. H.
1. I..
7.44 7.44
i Ii
7.32 7.5 1
I’) \\‘.I’.
I>. (i.
S.0 fi.0
7.60 7.60
fi.0 X.0
7..5H
7.49 7..51
7.60
7.67 7.67
7.X.5
IK(l
7.%
1X.0
II)
7.52
7.39
IO.0
12)
7.68 7.6”
7.85 7.70
IB.(l
(1)
7.60 7.39
7.64 7.61
X.,5 6.5
I”)
7.68 7.68
7.81 7.81
12.5
c:. ‘I.,
ill
I’. ‘;.
I
.1. I,.
II)
l-7.0
x.0 7..iO
7.30 7.,1x
I )
Awragf~:
of 400 400
1)lood. The control
IO.0 IO.0
7.33
7..56
7.0
7.41
7..58
7.0
7.ris 755
7.41 7.44
9.0
7.55
7.6 I
CJ.6
MOSM,
or more
290 MOSM ). the occurrence In addition.
14.0
7.65
(2)
osmolality
I “.(I I7.0
12)
( 1)
7.64
mg.
(compared
of hemolysis
per
samples
cent showed
H.0
to the
normal
of urea
was
a utilization
added
to aliquots
hour and the llrea loaded
samples
of 10.5 and 11 mg. l~r
addition
of urea
the acute
of
of normal
of 8.5 and 7.5 mg. per cent;
concluded
that
osmolalitv
is not surprising.
cent/hour.
did not duplicate
It was
the inhibition.
632
J. M. hIORGAN
Table Z.-Glucose Disappearance Blood-Leukocytes Excluded-Hematocrit
Azotemic PH’
PH’
G’
GZ
G.D.
7.50
7.50
152
142
5.0
1.50
7.55
152
136
8.0
7.51
7.59
146
146
7.57
7.49
88
60
14.0
7.50
7.45
60
36
12.0
.___ B. C.
(1)
(2)
0. T.
(1) (2)
C. H.
P. s.
(1)
(1) (2)
H. C.
Q. W.
C. M.
J. J.
(1)
7.48
7.56
114
100
7.0
7.63
110
90
10.0
7.56
7.57
162
144
9.0
7.59
7.57
162
148
7.0
7.5.4
7.57
164
142
11.0
7.50
7.65
90
64
13.0
7.59
7.65
104
78
13.0
7..59
7.71
114
10.1
5.5
7.59
7.62
115
!I6
7.58
7.62
112
90
11.0
7.59
7.62
112
R8
12.0
(1)
7.4x
7.46
122
9x
12.0
7.42
122
9G
1:z.o
7.5x
7.58
!I4
13.0
7.fil
7.6R
XX
14.5
7.51
7.52
:34
i.5
7.51
7.52
:34
7.5
7.411
X(1
6.0
i.60
7x
7.0
7.41 7.4
J. Mel
(1)
glomerulo-
nephritis
72
184
Postoperative
x2
Acute
renal
lowing 100
Chronic
failure
fol-
pancreatitis pyelonephritis
67
237
Pyelonephritis
and
nephrosclerosis 77
Nephronclerosin
39
124
Nephmscleros:s
Post-transfusion
142
116
14.0
i.52
136
100
1x.0
acute
7.50
7.58
144
11x
13.0
recovery
7.48
7.49
72
61
10.5
7.48
7.44
72
50
11.0
7.52
7.56
94
IO.3
III m!z.
re-
142
7.53
nitrogen
acute
nal failure
7.50
Average: urea
Diagnosis Chonric
7.50
(2)
*Blood
1
40 Per Cent
BUN* ____104
!)..5
7.50
(1)
Rate
0
7.46
(1)
AND R. E. MORGAN
renal
acute failure,
phase
per CenL.
DIscussroN
Glucose utilization has been incompletely studied in erythrocytes from uremic persons, but was stated by Rees I” to be normal until severe metabolic acidosis occurs. As shown above, our findings would agree wth this. HOWever, it has been shown that decreased red cell survival time is characteristic of late uremia,7 and it is generally agreed that young cells metabolize glucose more rapidly than aged cells. li In fact, the glycolytic rate in hemolytic disease has been considered a rough index of mean cell age by Hollingsworth.12 Therefore, with the young cell population present in late uremia. a high rate of glucose utilization would be expected. The observation of a normal rate of glycolysis in itself suggests an impairment. Our preliminary attempts to demonstrate augmentation of glycolysis in uremic cells on incubation in normal ultrafiltrate were obscured by the occurrence of hemolysis. In the present study, the effects of uremic ultrafiltrate on normal erythrocytes were
633 Table 3.-Glucose Disappearance LNormal Erythrocytes in Ultrafiltrate-Hematocrit I.
Normul cells-normal 7..50 7.46: 7.65 7.55 7.3 6s 7.46 7.U
40 Per Cent
ultrufiltrote 7.54 7.50 7.48 7.58 7.84 7.!53 7.58
ultrufltrate 7.4<5 7.41 7.Fi6 7.50 7.66 7.54 7.56 7.57 7..57
751 T.-G5 7.67 7.M 7.U 7.30 7.52 7.x
18..5 6.0 6.0
i.30
172 17.5 185 IHO 1.58 140 139 15’3 l-l2
10.6
IO4X
:3.7 3.3 17.2” 6.5 12.0” 6.3 3.7
5:3 76 ($2 66 67 57 31
4.0
14
Average: “Cclla from a patient 20 mg. pvr cmt/hr. studied
to circum\,ent
on ervthrocyte The
rate
The
this
as a cause
demonstrated
that within
33
6.‘3
55%
vrrii wllosc~ norn~i~l 11tilization wxh hi&
It would
glycolysis
seem
has been
of controlled
of the
the
observed
lel~el of the
wide
of initial
3.0
that
an inhibitorv
at
effect
is present.
maintenance
of glycoIysis centration
this problem.
gIycolysis of erythiocvte
changes.“’ eludes
with polycythemia
Control
11-l 121 86 108 103 100 100
ti.0 8.5 16.7 LO.*5
133 144 136 144 140 136 138
Av~gt:
II. A’orvral cdl~-Azotemic
7
G.D.
G’
pH’
PHI
Rate
pH
in the
to be sensitive
in the
abo\re incubations
differences.
blood
Previous does
glucose
ranges. i4 Therefore,
~ILICOS~
shown
above
the
groups
limited should
data
not
effect
variation not effect
to pH CAY-
have
also
the
ratt>
in COW the
rate
of glycolysis. Previous
imestigators
ha1.e sllggested
zation
in experimental
given,
acidosis
as a cause
would
suggest
that
The observed changes
such
inhibition
in the erythrocyte a decreased
erythrocytc gators.
erythrocyte
glucose
utili-
in the dog.‘“~‘” However. since pH’s were not of the defect was not excluded. The present data
an impairment
mav
be present
in the
absence
c)f
in man.
acidosis
or (2)
decreased
uremia
There
of simple Inaintained
rate
membrane
could
on theoretical
membrane, of gIycoIvsis.
has been
grounds
decreasing
The extensively
as maintained
by LeFevre’”
bv h4auwe”
and others:
However.
glucose
to:
(1)
to gh~cose:
movement of gIucose studied by a number
seems to be some doubt as to whether
diffusion,
be attributed
its permeability
across the of investi-
entry is the result
or of facilitated all investigators
diffusion, ;LS agree that it
634
1. M.
MORGAX
AND R. E. MORGAN
is very rapid at wide ranges of plasma glucose concentration. Therefore, it seems unlikely that this would be rate limiting in erythrocyte glycolysis. However, Allison and Burn” have pointed out that the non-nucleated erythrocyte cannot produce enzymes and therefore must depend on the supply inherited from the nucleated precursor. Since active energy production through glycolysis is necessary to maintain the functional erythrocytez-4 the inhibition or depletion of essential enzymes below critical levels could lead to cell destruction. Previous work from this laboratory has suggested that uremic ultrafiltrate has an inhibitory effect on the enzyme lactic dehydrogenase. In Attempts have also been made to demonstrate an ef?ect on aldolasc but no inhibition was apparent.“” Therefore, much more work needs to be done to establish the validity of such a possibility. REFERENCES I. Murphy, J. R.: Erythrocyte J. Lab.
& Clin.
hlod.
metabolism. 55:286, 1960.
2. hlaizcls, M.: Factors in the active transport of cations. Am. J. Physiol. 112: 59,
COlySlS
Hemat.
Blood
1:131,
9:227,
of the red cell.
1954.
erythrocytes.
Science
124:484,
ficiency
in hereditary
hemolytic
anemia.
1962. 7. Joske, R. A., hlcAlistcr, J, M., and Prankerd, T. A. J.: Isotope investigations of red cell production and destruction
in
Clin.
15:511,
Sci.
chronic
renal
disease.
1956.
G. M., Mackler, H., and Ammentorp, utilization of glucose
B., Graubarth, P. A.: Rate of in erythrocytes
and leucocytes. Am. J. Physiol. 172: 295, 1953. 9. Somogyi, J.: Determination of blood sugar. J. Biol. Chem. 160:69, 1945. 10. Rees, S. B.: In Schreiner, G. E. (Ed.): Transactions of the Am. Sot. for Artificial Internal Organs, Vol. IV, p. 202. 11. Allison, A. C., and Burn, G. P.: Enzyme activity as a function of age in the
Effects glucose
of
Chem.
19%.
J.
in utiliza-
Physiol.
E.,
of erythrocytes
F.:
Low-
phosphate
glutathione
following
J. Lab.
blood.
1947.
and Jones,
take and reduced
anti
172:301,
in hulllan
169:493,
ered glucose utilization,
phrectomy.
Lab.
in erythrocytes
Am.
E.
gly-
J.
B., and Guest,
of acidosis
R. M. Glycolysis
15. Muirhead,
Hemat.
diseas;e.
45:920,
tion
J.
Erythrocytc
H., Mackler,
J. Biol.
and cle-
19:267,
W.:
G. RI.:
14. Bird,
nonspherocytic Blood
Med.
leucocytrs. 1953.
1956.
K. R., Valentin, W. N., S.: Pyruvate kinase (PK)
J.
.
Brit.
in hemolytic
13. Graubarth,
1955.
5. Carson, P. E., Flanagan, G. L., Iches, C. E., and Alving, A. S.: Enzymatic primaquin sensitive deficiency in 6. Tanaka, Miwa,
.
& Clin. metabolism of A review. Brit.
4. Ponder, E.: Present concepts structure of tthe mammalian
8. Guest,
12. Hollingsworth,
1951.
3. Prankerd, T. A. J.: The the human erythrocyte. J.
human erythrocytc. 1:291, 195.5.
up-
content
bilateral
& Clin.
ne-
Med.
51:
49, 1958. 16. -,
and -:
Lowered
glucose
utilization,
phosphate uptake and reduced glutathione of erythrocytes following uretcrocaval Exp. 17. Mawe, into
Biol. R. D.: the
Camp. 18. LcFevre,
anastamosis. 102:351, The
human Physiol.
P. D.:
red blood
cell:
Proc.
Sot.
1959. diffusion
red
cell.
47: 177, Sugar
of glucose J.
transport
structure
Cell.
&
1956. in the
activity
re-
lationships in substrates and antagonists. Pharm. Rev. 13:39, 1961. 19. Morgan, J. M., Morgan, R. E., and Thomas, G. E.: Inhibition of lactic dehydrogenase uremic blood. 1963. 20. Unpublished
by ultra-filtrate of Metabolism 12: 1051,
observations.
REMK:
I1IETAHOLITES
ON EHYTHROC:Y’L‘E GLYC:OLYSIS
Jean Al. hforgun, M.D., Assistant Professor of Medicine, Medkwl College of Alubumn, Birmingham, Alabama, and Assista& Chief, Ale,dical Sewice, Veterans Administration Ilospital, Birmingham, Alu. Present address: Medical College of South Carolina, 80 Barre St., Charleston, S. C. Hobert E. Morgan, D.D.S., M.Sc., Assistant Professor of Dentistry, School of Dentistry, Birmingham, Alabama, und Clinical lwestigutor, Veterans Administration Ilospital, Birmitlglwm, Ala. Present Address: Medical College of Sorrfh Carolina, 80 Barre St.. Charleston, S. C.
Attempts
have
been
JEAN
made
effect of uremic metabolites rocyte glycolysis. In blood persons, glucose filtrate
E
the rate is normal. of uremic
XTENSIVE
M. MORGAS AND ROBERTE. MORGAN to study
the
upon erythfrom uremic
of disappearance of However, the ultrablood
produces
inhibi-
tion
of
glucose
erythrocytes. in
uremic
The blood
utilization failure may
by
normal
to observe be
due
to
this the
younger erythrocyte population. The mechanism and possible significance of this inhibition
remain
to be shown.
STUDIES ha\xs ken reported in recent vears concerning essential metabolic processes in the erythrocvte. Erythrocvte metabolism is entirely anaerobic.] The energy derived from glycolysis and stored ;LS the “pump” which exadenosine triphosphate (ATP) lr, ‘: necessary to maintain trudes sodium and water from the cell and retains the necessary high potasGllrn concentration.“,” The maintenance of the cell membrane is also energ\’ rccpiring4 Recent studies of certain congenital hemolvtic anemias ha\rct illustrated the ervthrocvte fragilitv which occurs as a result of glucose-fiphosphate dehvdrogenase; and pyruvate kinasc” deficiency, both leading to impaired glycolvsis. The anemia of severe rlremia has been shown to 1)(x hemolvtic in na&e,’ which suggests the possibility of a metabolic defect. The present study was therefore designed to determine: (1) the rate at whiclI glucose is utilized by erythrocytes from uremic persons under varied conditions: and (2) the effect of ultrafiltrate of blood from llremic persons on normal red blood c,ell metabolism.
630
J.M.
MORGANAND R.E. MORGAN
after 2 hours of incubation. Each incubation was carried out in duplicate. The rate of disappearance of glucose was assumed to be the erythrocyte utilization rate. To prepare the ultrafiltrates, sera from normal and uremic persons was collected and frozen until processed. Each serum was placed in a boiled cellophane membrane and centrifuged in the Tori-Bara apparatus at 2700 RPM for 16-24 hours at 5” C. The ultrafiltrate was kept frozen until utilized for study. At that time 100 mg. per cent glucose was added
and the ultrafiltrate
rected
to 40 per cent and incubations
was mixed
with
fresh
were
carried
erythrocytes.
The
hematocrit
was
cor-
out as indicated.
RESULTS
The glucose utilized by erythrocytes from 10 normal subjects, determined in native plasma, was measured on several occasions. The average for the group of 17 duplicate determinations was 9.6 mg. per cent/hour, with limits of 618 mg. per cent/hour. The data are summarized in table 1. The final pH determinations \,aried from 7.44 to 7.85. The initial blood sugar concentrations were between 34 to 102 mg. per cent with an average of 65 mg. per cent. The rate of disappearance of glucose in blood from azotemic persons was not significantly different from that in the normal group. The average for this group of 9 persons, several studied on more than one occasion, was 10.3 mg. per cent. Initial blood-sugar concentrations averaged 113 mg. per cent, with a range of 34 to 148 mg. per cent. The data are summarized in table 2. Three of the patients studied suflered from acute renal failure and 6 had longstanding moderate to severe uremia. The blood urea nitrogen (BUN) concentrations varied from 39 to 237 mg. per cent, and all but 1 patient had some degree of anemia. No correlation was apparent between either the BUN 01 creatinlne concentration and the rate of glucose utilization. When erythrocytes from healthy donors were incubated in ultrafiltratc from the same or other healthy persons, the rate of glucose utilization was not significantly different from that of the erythrocytes in their native plasma (specified as “control”). The rate of utilization averaged 104 per cent of the control, with all initial blood sugars between 135 and 144 mg. per cent and the final pH’s varying from 7.48-7.84. The ultrafiltrate from azotemic persons was distinctly inhibitory by comparison, with the rate of glucose disappearance averaging only 55 per cent of the control. The data are summarized in table 3. The initial blood sugars varied from 139 to 185 mg. per cent and the final pH from 7.4 to 7.66 mg. per cent. Three of the ultrafiltrates were obtained from patients shown in table 2 (O.T., P.S. and B.C.). Five others were from selrere chronic uremics and one was from an acute renal failure patient just prior to hemodialysis. Clinically, the patients in this group were more symptomatic as a result of renal failure than most of the patients summarized in table 2. An attempt was made to incubate the erythrocytes from uremic persons in normal ultrafiltrate. However, the results were highly variable. In 4 trials, the rate of disappearance of glucose was 36, 107, 121 and 283 per cent of that in the native plasma. Marked discoloration of the ultrafiltrate was noted after
centrifllgation
the cause
of the
of the
first
low utilization.
sample.
This
Since
severely
suggested uremic
that blood
hemolysis may
have
was an
LlRE:JfI(:
METABOLITES
6.31
C:LU(:OLYSlS
Table l.-Glucose Disappearance Rate Blood-Leukocytes Excluded-Hemutocrit 40 Per Cenf
Normal H. Y.
ON EHTTHRO~:\iTE
PH’
*H’
(1)
7.50 7.60
7.31 756
15 .5 1Z!..li
(2)
7.48
7..50 7.65
8.0 x.0
7.53 I31
G.U.
10.0
7.49 7.48
7.48
10.0
11. H.
i I)
7.34 753
7..54 7..55
7.0 7.r,
J.\\.
il)
7.*57 7.57
7.62
12.0 12.0
I. H.
1. I..
7.44 7.44
i Ii
7.32 7.5 1
I’) \\‘.I’.
I>. (i.
S.0 fi.0
7.60 7.60
fi.0 X.0
7..5H
7.49 7..51
7.60
7.67 7.67
7.X.5
IK(l
7.%
1X.0
II)
7.52
7.39
IO.0
12)
7.68 7.6”
7.85 7.70
IB.(l
(1)
7.60 7.39
7.64 7.61
X.,5 6.5
I”)
7.68 7.68
7.81 7.81
12.5
c:. ‘I.,
ill
I’. ‘;.
I
.1. I,.
II)
l-7.0
x.0 7..iO
7.30 7.,1x
I )
Awragf~:
of 400 400
1)lood. The control
IO.0 IO.0
7.33
7..56
7.0
7.41
7..58
7.0
7.ris 755
7.41 7.44
9.0
7.55
7.6 I
CJ.6
MOSM,
or more
290 MOSM ). the occurrence In addition.
14.0
7.65
(2)
osmolality
I “.(I I7.0
12)
( 1)
7.64
mg.
(compared
of hemolysis
per
samples
cent showed
H.0
to the
normal
of urea
was
a utilization
added
to aliquots
hour and the llrea loaded
samples
of 10.5 and 11 mg. l~r
addition
of urea
the acute
of
of normal
of 8.5 and 7.5 mg. per cent;
concluded
that
osmolalitv
is not surprising.
cent/hour.
did not duplicate
It was
the inhibition.
632
J. M. hIORGAN
Table Z.-Glucose Disappearance Blood-Leukocytes Excluded-Hematocrit
Azotemic PH’
PH’
G’
GZ
G.D.
7.50
7.50
152
142
5.0
1.50
7.55
152
136
8.0
7.51
7.59
146
146
7.57
7.49
88
60
14.0
7.50
7.45
60
36
12.0
.___ B. C.
(1)
(2)
0. T.
(1) (2)
C. H.
P. s.
(1)
(1) (2)
H. C.
Q. W.
C. M.
J. J.
(1)
7.48
7.56
114
100
7.0
7.63
110
90
10.0
7.56
7.57
162
144
9.0
7.59
7.57
162
148
7.0
7.5.4
7.57
164
142
11.0
7.50
7.65
90
64
13.0
7.59
7.65
104
78
13.0
7..59
7.71
114
10.1
5.5
7.59
7.62
115
!I6
7.58
7.62
112
90
11.0
7.59
7.62
112
R8
12.0
(1)
7.4x
7.46
122
9x
12.0
7.42
122
9G
1:z.o
7.5x
7.58
!I4
13.0
7.fil
7.6R
XX
14.5
7.51
7.52
:34
i.5
7.51
7.52
:34
7.5
7.411
X(1
6.0
i.60
7x
7.0
7.41 7.4
J. Mel
(1)
glomerulo-
nephritis
72
184
Postoperative
x2
Acute
renal
lowing 100
Chronic
failure
fol-
pancreatitis pyelonephritis
67
237
Pyelonephritis
and
nephrosclerosis 77
Nephronclerosin
39
124
Nephmscleros:s
Post-transfusion
142
116
14.0
i.52
136
100
1x.0
acute
7.50
7.58
144
11x
13.0
recovery
7.48
7.49
72
61
10.5
7.48
7.44
72
50
11.0
7.52
7.56
94
IO.3
III m!z.
re-
142
7.53
nitrogen
acute
nal failure
7.50
Average: urea
Diagnosis Chonric
7.50
(2)
*Blood
1
40 Per Cent
BUN* ____104
!)..5
7.50
(1)
Rate
0
7.46
(1)
AND R. E. MORGAN
renal
acute failure,
phase
per CenL.
DIscussroN
Glucose utilization has been incompletely studied in erythrocytes from uremic persons, but was stated by Rees I” to be normal until severe metabolic acidosis occurs. As shown above, our findings would agree wth this. HOWever, it has been shown that decreased red cell survival time is characteristic of late uremia,7 and it is generally agreed that young cells metabolize glucose more rapidly than aged cells. li In fact, the glycolytic rate in hemolytic disease has been considered a rough index of mean cell age by Hollingsworth.12 Therefore, with the young cell population present in late uremia. a high rate of glucose utilization would be expected. The observation of a normal rate of glycolysis in itself suggests an impairment. Our preliminary attempts to demonstrate augmentation of glycolysis in uremic cells on incubation in normal ultrafiltrate were obscured by the occurrence of hemolysis. In the present study, the effects of uremic ultrafiltrate on normal erythrocytes were
633 Table 3.-Glucose Disappearance LNormal Erythrocytes in Ultrafiltrate-Hematocrit I.
Normul cells-normal 7..50 7.46: 7.65 7.55 7.3 6s 7.46 7.U
40 Per Cent
ultrufiltrote 7.54 7.50 7.48 7.58 7.84 7.!53 7.58
ultrufltrate 7.4<5 7.41 7.Fi6 7.50 7.66 7.54 7.56 7.57 7..57
751 T.-G5 7.67 7.M 7.U 7.30 7.52 7.x
18..5 6.0 6.0
i.30
172 17.5 185 IHO 1.58 140 139 15’3 l-l2
10.6
IO4X
:3.7 3.3 17.2” 6.5 12.0” 6.3 3.7
5:3 76 ($2 66 67 57 31
4.0
14
Average: “Cclla from a patient 20 mg. pvr cmt/hr. studied
to circum\,ent
on ervthrocyte The
rate
The
this
as a cause
demonstrated
that within
33
6.‘3
55%
vrrii wllosc~ norn~i~l 11tilization wxh hi&
It would
glycolysis
seem
has been
of controlled
of the
the
observed
lel~el of the
wide
of initial
3.0
that
an inhibitorv
at
effect
is present.
maintenance
of glycoIysis centration
this problem.
gIycolysis of erythiocvte
changes.“’ eludes
with polycythemia
Control
11-l 121 86 108 103 100 100
ti.0 8.5 16.7 LO.*5
133 144 136 144 140 136 138
Av~gt:
II. A’orvral cdl~-Azotemic
7
G.D.
G’
pH’
PHI
Rate
pH
in the
to be sensitive
in the
abo\re incubations
differences.
blood
Previous does
glucose
ranges. i4 Therefore,
~ILICOS~
shown
above
the
groups
limited should
data
not
effect
variation not effect
to pH CAY-
have
also
the
ratt>
in COW the
rate
of glycolysis. Previous
imestigators
ha1.e sllggested
zation
in experimental
given,
acidosis
as a cause
would
suggest
that
The observed changes
such
inhibition
in the erythrocyte a decreased
erythrocytc gators.
erythrocyte
glucose
utili-
in the dog.‘“~‘” However. since pH’s were not of the defect was not excluded. The present data
an impairment
mav
be present
in the
absence
c)f
in man.
acidosis
or (2)
decreased
uremia
There
of simple Inaintained
rate
membrane
could
on theoretical
membrane, of gIycoIvsis.
has been
grounds
decreasing
The extensively
as maintained
by LeFevre’”
bv h4auwe”
and others:
However.
glucose
to:
(1)
to gh~cose:
movement of gIucose studied by a number
seems to be some doubt as to whether
diffusion,
be attributed
its permeability
across the of investi-
entry is the result
or of facilitated all investigators
diffusion, ;LS agree that it
634
1. M.
MORGAX
AND R. E. MORGAN
is very rapid at wide ranges of plasma glucose concentration. Therefore, it seems unlikely that this would be rate limiting in erythrocyte glycolysis. However, Allison and Burn” have pointed out that the non-nucleated erythrocyte cannot produce enzymes and therefore must depend on the supply inherited from the nucleated precursor. Since active energy production through glycolysis is necessary to maintain the functional erythrocytez-4 the inhibition or depletion of essential enzymes below critical levels could lead to cell destruction. Previous work from this laboratory has suggested that uremic ultrafiltrate has an inhibitory effect on the enzyme lactic dehydrogenase. In Attempts have also been made to demonstrate an ef?ect on aldolasc but no inhibition was apparent.“” Therefore, much more work needs to be done to establish the validity of such a possibility. REFERENCES I. Murphy, J. R.: Erythrocyte J. Lab.
& Clin.
hlod.
metabolism. 55:286, 1960.
2. hlaizcls, M.: Factors in the active transport of cations. Am. J. Physiol. 112: 59,
COlySlS
Hemat.
Blood
1:131,
9:227,
of the red cell.
1954.
erythrocytes.
Science
124:484,
ficiency
in hereditary
hemolytic
anemia.
1962. 7. Joske, R. A., hlcAlistcr, J, M., and Prankerd, T. A. J.: Isotope investigations of red cell production and destruction
in
Clin.
15:511,
Sci.
chronic
renal
disease.
1956.
G. M., Mackler, H., and Ammentorp, utilization of glucose
B., Graubarth, P. A.: Rate of in erythrocytes
and leucocytes. Am. J. Physiol. 172: 295, 1953. 9. Somogyi, J.: Determination of blood sugar. J. Biol. Chem. 160:69, 1945. 10. Rees, S. B.: In Schreiner, G. E. (Ed.): Transactions of the Am. Sot. for Artificial Internal Organs, Vol. IV, p. 202. 11. Allison, A. C., and Burn, G. P.: Enzyme activity as a function of age in the
Effects glucose
of
Chem.
19%.
J.
in utiliza-
Physiol.
E.,
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F.:
Low-
phosphate
glutathione
following
J. Lab.
blood.
1947.
and Jones,
take and reduced
anti
172:301,
in hulllan
169:493,
ered glucose utilization,
phrectomy.
Lab.
in erythrocytes
Am.
E.
gly-
J.
B., and Guest,
of acidosis
R. M. Glycolysis
15. Muirhead,
Hemat.
diseas;e.
45:920,
tion
J.
Erythrocytc
H., Mackler,
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W.:
G. RI.:
14. Bird,
nonspherocytic Blood
Med.
leucocytrs. 1953.
1956.
K. R., Valentin, W. N., S.: Pyruvate kinase (PK)
J.
.
Brit.
in hemolytic
13. Graubarth,
1955.
5. Carson, P. E., Flanagan, G. L., Iches, C. E., and Alving, A. S.: Enzymatic primaquin sensitive deficiency in 6. Tanaka, Miwa,
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& Clin. metabolism of A review. Brit.
4. Ponder, E.: Present concepts structure of tthe mammalian
8. Guest,
12. Hollingsworth,
1951.
3. Prankerd, T. A. J.: The the human erythrocyte. J.
human erythrocytc. 1:291, 195.5.
up-
content
bilateral
& Clin.
ne-
Med.
51:
49, 1958. 16. -,
and -:
Lowered
glucose
utilization,
phosphate uptake and reduced glutathione of erythrocytes following uretcrocaval Exp. 17. Mawe, into
Biol. R. D.: the
Camp. 18. LcFevre,
anastamosis. 102:351, The
human Physiol.
P. D.:
red blood
cell:
Proc.
Sot.
1959. diffusion
red
cell.
47: 177, Sugar
of glucose J.
transport
structure
Cell.
&
1956. in the
activity
re-
lationships in substrates and antagonists. Pharm. Rev. 13:39, 1961. 19. Morgan, J. M., Morgan, R. E., and Thomas, G. E.: Inhibition of lactic dehydrogenase uremic blood. 1963. 20. Unpublished
by ultra-filtrate of Metabolism 12: 1051,
observations.
REMK:
I1IETAHOLITES
ON EHYTHROC:Y’L‘E GLYC:OLYSIS
Jean Al. hforgun, M.D., Assistant Professor of Medicine, Medkwl College of Alubumn, Birmingham, Alabama, and Assista& Chief, Ale,dical Sewice, Veterans Administration Ilospital, Birmingham, Alu. Present address: Medical College of South Carolina, 80 Barre St., Charleston, S. C. Hobert E. Morgan, D.D.S., M.Sc., Assistant Professor of Dentistry, School of Dentistry, Birmingham, Alabama, und Clinical lwestigutor, Veterans Administration Ilospital, Birmitlglwm, Ala. Present Address: Medical College of Sorrfh Carolina, 80 Barre St.. Charleston, S. C.