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Comparative effects of pyridoxine, riboflavin and thiamine on linoleate utilization in rats
BIOCHIMICA ET BIOPHYSICA
388
SHORT
ACTA
COMMUNICATIONS
BBA 53120
Comparative)effects utilization
The evidence been derived mentation acid
from
on linoleate
pyridoxine
the same
utilization
without
of both
linoleate
essential “it
that
pyridoxine
could
found
and pyridoxine,
supplementation higher
linoleate
that level
increase
present
on recovery the effect previous
fatty
depletion
of essential
control
adequate pyridoxine into
fatty
group flavin
or pyridoxine, of linoleate tube.
fatty
The third
source stomach
plus (80%
diet
of
growth.
SCHEIER
essential
fatty
acid
for 6 days produced
than did supplementa-
Acta,
of a deficiency
report,
from essential
fatty
acid. After
we have tested
in rats fed linoleate
but containing
vitamins6.
lipid
The
received
the
supplement
diet was restricted
basal
second
the daily
food
to the average
and food intake
of
with ribooil was the
and was given
intake daily
sub-
supplement
Safflower
analysis)
was
was subdivided The
plus a daily
of linoleate.
dietary diet
of riboflavin,
the same diet supplemented
chromatography period,
analyses.
as before,
45-50
with riboflavin
each group
stopped,
for liver
as
after
pyridoxine,
was the only
growth diet
Growth
which
the exception
subgroup
137 (1967) 388-390
to the vitamin
acid depletion,
by the rat, with
was sacrificed
diet.
Up
vitamin.
oil (z%),
of fat-soluble
required
daily
of whether
of another
In the present
riboflavin
coconut
In the supplementation
the vitamin-supplemented
the question
of pyridoxine.
arachidonate
lacking
by gas-liquid
rats fed the vitamin-deficient Riochim. Biophys.
results
conversion
of male rats (ZI days old, Long-Evans,
the same vitamin-deficient
100 mg linoleate.
tissue
of both
effect
acid and either
absorption
One subgroup
received
WIT-
in rats fed both
was fed the same basal diet supplemented
nutrients7
and essential
a specific
on recovery
Hydrogenated
to improve
in all other
3 groups.
fed
the rats
similar
in arachidonate greater
with
rats
in the conversion
and linoleate
on the effect
one group
group
pyridoxine.
was added
than
in the
AND CONIGLIO raised
phospholipid
casein-sucrose
g) was fed a 20%
fat,
fatty
Consequently,
obtained
arachidonate
acid depletion.
in liver
In the first experiment,
but lacking
rat
is involved
depleted
essential
supplementation,
involved
from
pyridoxine
was
and thiamine
by the changes
and a second
whole
pyridoxine.
increase
phospholipid
of no studies
essential
of riboflavin
measured
of liver
of both
has
supple-
rats supplemented
AND CONICLIO*
resulted
with
arachidonate
we know
from
have
Before
specifically
in rats previously
of KIRSCHMAN
in tissue
time,
per
pyridoxine
not
of pyridoxine
alone.
The interpretation the
arachidonate acid and
that
was
AND WILLIAMS~ a significantly
fatty
since the greater
and linoleate
that
KIRSCHMAN
pyridoxine
to arachidonate
tion with
linoleate
acid metabolisml~”
depleted
observed
pyridoxine.
appears
to arachidonate”.
suggested
more
fatty
the effect
by rats previously
of linoleate wrote,
tested
AND HOLMAN~ had
TEN AND HOLMAN
in essential
which
and linoleate
amount
of linoleate
experiments
WITTEN
had been depleted
but
riboflavin and thiamineon
for a role of pyridoxine
chiefly
and pyridoxine.
both
of pyridoxine,
in rats
by
of the rats fed
food
intake
data are given
of the
in Table
I.
SHORT
389
COMMUNICATIONS
TABLE
I
EFFECT OF DIETARY TREATMENTS
11’0 of vats
Treatment
Experiment
ON BODY
Initial weight
WEIGHT
Double depletion
(g)
time (days)
AND
FOOD
INTAKE
Weight after EFA addition
Weight at start of EFA addition
fg)
(g)
Food eaten duving EFA addition (g)
I
-B,
-EFA
7
45
28
96
-B, +B, -B,
+EFA +EFA -EFA
7 7 7
46 46 46
28 28 21
97 95 7’
1 +7 -
43 43
-B, +B,
+EFA +EFA
7 7
45 46
21 21
69 7o
i- 5 +I4
36 36
Experiment
-
2
-B,
-EFA
6
47
19
67
-
-B, +B, -B,
+EFA +EFA -EFA
6 6 6
5o 48 47
19 19 7
7o 67 7o
+1 +14
34 34 -
-B, +B,
+EFA +EFA
6 6
49 49
7 7
69 67
+9 +19
41 41
-._ Abbreviations flower oil.
TABLE LIVER
:
B,,
pyridoxine
___. B,,
;
riboflavin;
B,, thiamine;
EF.4,
essential fatty
-
acid e.g. saf-
II PHOSPHOLIPID
ARACHIDONATE
Means and standard
Liver weight Phospholipid (mg/g liver) (9)
Treatment
E,@erimelzt
VALUES
error.
Phospholipid (weight %)
arachidonate (mglliuer)
(mglg liver)
I
--IS,
-EFI
7*
4.77
27.4 -h 1.1
13.7 & 0.4
12.8 & 0.8
2.7 f
-B, +B,
+EF_k +EFr\
7 7
4.65 4.30
28.6 * I.2 33.6 & 1.1
26.1 + 1.8 28.9 + 0.6
23.9 I 0.3 30.2 + 1.5
5.5 zt 0.6 7.0 i 0.3
-B, -B,
-EFh AEFA
7 7
2.99 3.26
30.4 & 0.8 29.7 i I.5
17.1 & 1.5 28.5 & 1.4
II.0 _k 0.9 20.0 t I.9
4.0 i 0.6 6.2 17 0.5
+B,
+EFA
7
3.72
30.1 =k 0.7
26.8 *
1.0
21.5 & 1.1
5.8 k 0.3
Expevime??l
0.2
2
-B, -B, +B,
-EFh +EFX +EFA
6 6 6
2.85 3.07 2.94
34.4 i 0.4 33.9 * I.2 38.0 & 0.6
15.2 & 0.9 26.0 f 1.1 30.4 & 0.6
10.7 & 0.7 19.3 + 1.2 24.4 * 1.0
3.8 * 6.3 i 8.3 :
-B, -BI +B,
-EFX +EF,\ --EFh
6 6 6
4.05 3.34 3.10
28.7 + 2.4 30.3 f 2.8 36.0 3 1.1
12.8 -& 0.6 28.4 * 0.9 32.2 * 1.2
10.7 + 0.9 20.4 * 1.5 25.9 i 1.5
2.6 f 0.2 6.1 + 0.3 8.3 & 0.2
Abbreviations: oil. * Number
B,, pyridoxine;
B,, riboflavin;
B,, thiamine;
EFA, essential fatty.acid
0.2 0.4 0.2
e.g. safflower
of rats per group.
After 6 days of linoleate suppementation, the rats were sacrificed by decapitation without anesthesia, Liver lipids were extracted, fractionated and fatty acids determined as previously described798. In the second experiment, the same dietary procedures were used, except that thiamine depletion, rather than riboflavin depletion, was produced. Pyridoxinedepleted
rats again were the controls. Biochim. Biophys. Acta, 137 (1967) 388-390
SHORT COhfMUh'ICATIONS
390
Liver phospholipid and phospholipid arachidonate values are given in Table II. In the first experiment, the degree of arachidonate depletion was similar in both vitamin-deficient groups at the start of the supplementation period (lines I and 4). In the group previously depleted of both pyridoxine and essential fatty acid, supplementation with pyridoxine and safflower oil produced a significantly higher level of arachidonate than did supplementation with safflower oil only.The difference in arachidonate was significant when expressed as mg/liver or as mg/g liver. The difference in weight o/0 was not significant. In the group previously depleted of riboflavin and essential fatty acid, supplementation with riboflavin and safflower oil, however, produced no greater increase in arachidonate than did supplementation with safflower oil alone. Therefore, riboflavin appeared to have no effect on arachidonate level. In the second experiment, the degree of arachidonate depletion was again similar in both vitamin-deficient groups at the start of supplementation (lines 7 and IO). In the group previously depleted of both pyridoxine and essential fatty acid, supplementation with pyridoxine and safflower oil produced a significantly higher level of arachidonate than did supplementation with the oil alone, in agreement with the first experiment. The differences were significantly greater when expressed as weight %, as mg/liver, or as mg/g liver. In the group depleted of thiamine and essential fatty acid, supplementation with thiamine and safflower oil also produced a higher level of arachidonate (weight o/O,mg/liver, mg/g liver) than did supplementation with safflower oil. Consequently, no evidence for a specific effect of pyridoxine in the conversion of linoleate to arachidonate can be inferred from this type of experiment in vivo since thiamine produced similar changes. Experiments ilz vitro may give more information on the role of pyridoxine or thiamine in arachidonate formation. GOSWAMI AND CONIGLIO~ recently observed a stimulatory effect of pyridoxal phosphate on the incorporation of [r-14]acetyl CoA into arachidonate and docosahexaenoate by liver microsomes. This study was aided by U.S. Public Health Service, Grant AM 7753. Department of Nutritional
Sciences
Ufiiversity of California Berkeley, Calif, (U.S.A.)
M.A. WILLIAMS D.J. MCINTOSH I. HINCENBERGS K.T.TAMAI
I J. F. MUELLER, T~itaminsHormones, 22 (1964) 787. 2 J. F. MEAD, Ann. Rev. Biochem.. 32 (1963) 241. 3 P. W. WITTEN AND R. T. HOLMAN, Arch. Biochem. Biophys., 41 (1952) 266. 4 J. C. KIRSCHMAN AND J. G. CONIGLIO, J.Biol. Chem., 236 (1961) 2200. =,G. E. SCHEIER AND M. A. WILLIAMS. Biochem. I.. 42 (146~1 422. 6 A. R. HANDS, N. S. SUTHERLAND A&W. BAR;L%,%&&~. J., 94 (1965) 279. 7 M. A. WILLIAMS, D. 1. MCINTOSH AND I. HINCENBERGS, 1. iVutvition, 88 (1966) rg-, s R. L. LYMAN, R. OS&ALD, P. BOUCHARD AND A. SHANNON, Biochem. J.,‘gS (;966) 4.38. g .4. GOSWAMI END J. G. CONIGLIO, Abstr., 7th Intern. Congr. Nutrition, (1966) 253.
Received
October
Biochim. Biophys.
rrth,
1966
Acta, 137 (1967) 388-390
388
SHORT
ACTA
COMMUNICATIONS
BBA 53120
Comparative)effects utilization
The evidence been derived mentation acid
from
on linoleate
pyridoxine
the same
utilization
without
of both
linoleate
essential “it
that
pyridoxine
could
found
and pyridoxine,
supplementation higher
linoleate
that level
increase
present
on recovery the effect previous
fatty
depletion
of essential
control
adequate pyridoxine into
fatty
group flavin
or pyridoxine, of linoleate tube.
fatty
The third
source stomach
plus (80%
diet
of
growth.
SCHEIER
essential
fatty
acid
for 6 days produced
than did supplementa-
Acta,
of a deficiency
report,
from essential
fatty
acid. After
we have tested
in rats fed linoleate
but containing
vitamins6.
lipid
The
received
the
supplement
diet was restricted
basal
second
the daily
food
to the average
and food intake
of
with ribooil was the
and was given
intake daily
sub-
supplement
Safflower
analysis)
was
was subdivided The
plus a daily
of linoleate.
dietary diet
of riboflavin,
the same diet supplemented
chromatography period,
analyses.
as before,
45-50
with riboflavin
each group
stopped,
for liver
as
after
pyridoxine,
was the only
growth diet
Growth
which
the exception
subgroup
137 (1967) 388-390
to the vitamin
acid depletion,
by the rat, with
was sacrificed
diet.
Up
vitamin.
oil (z%),
of fat-soluble
required
daily
of whether
of another
In the present
riboflavin
coconut
In the supplementation
the vitamin-supplemented
the question
of pyridoxine.
arachidonate
lacking
by gas-liquid
rats fed the vitamin-deficient Riochim. Biophys.
results
conversion
of male rats (ZI days old, Long-Evans,
the same vitamin-deficient
100 mg linoleate.
tissue
of both
effect
acid and either
absorption
One subgroup
received
WIT-
in rats fed both
was fed the same basal diet supplemented
nutrients7
and essential
a specific
on recovery
Hydrogenated
to improve
in all other
3 groups.
fed
the rats
similar
in arachidonate greater
with
rats
in the conversion
and linoleate
on the effect
one group
group
pyridoxine.
was added
than
in the
AND CONIGLIO raised
phospholipid
casein-sucrose
g) was fed a 20%
fat,
fatty
Consequently,
obtained
arachidonate
acid depletion.
in liver
In the first experiment,
but lacking
rat
is involved
depleted
essential
supplementation,
involved
from
pyridoxine
was
and thiamine
by the changes
and a second
whole
pyridoxine.
increase
phospholipid
of no studies
essential
of riboflavin
measured
of liver
of both
has
supple-
rats supplemented
AND CONICLIO*
resulted
with
arachidonate
we know
from
have
Before
specifically
in rats previously
of KIRSCHMAN
in tissue
time,
per
pyridoxine
not
of pyridoxine
alone.
The interpretation the
arachidonate acid and
that
was
AND WILLIAMS~ a significantly
fatty
since the greater
and linoleate
that
KIRSCHMAN
pyridoxine
to arachidonate
tion with
linoleate
acid metabolisml~”
depleted
observed
pyridoxine.
appears
to arachidonate”.
suggested
more
fatty
the effect
by rats previously
of linoleate wrote,
tested
AND HOLMAN~ had
TEN AND HOLMAN
in essential
which
and linoleate
amount
of linoleate
experiments
WITTEN
had been depleted
but
riboflavin and thiamineon
for a role of pyridoxine
chiefly
and pyridoxine.
both
of pyridoxine,
in rats
by
of the rats fed
food
intake
data are given
of the
in Table
I.
SHORT
389
COMMUNICATIONS
TABLE
I
EFFECT OF DIETARY TREATMENTS
11’0 of vats
Treatment
Experiment
ON BODY
Initial weight
WEIGHT
Double depletion
(g)
time (days)
AND
FOOD
INTAKE
Weight after EFA addition
Weight at start of EFA addition
fg)
(g)
Food eaten duving EFA addition (g)
I
-B,
-EFA
7
45
28
96
-B, +B, -B,
+EFA +EFA -EFA
7 7 7
46 46 46
28 28 21
97 95 7’
1 +7 -
43 43
-B, +B,
+EFA +EFA
7 7
45 46
21 21
69 7o
i- 5 +I4
36 36
Experiment
-
2
-B,
-EFA
6
47
19
67
-
-B, +B, -B,
+EFA +EFA -EFA
6 6 6
5o 48 47
19 19 7
7o 67 7o
+1 +14
34 34 -
-B, +B,
+EFA +EFA
6 6
49 49
7 7
69 67
+9 +19
41 41
-._ Abbreviations flower oil.
TABLE LIVER
:
B,,
pyridoxine
___. B,,
;
riboflavin;
B,, thiamine;
EF.4,
essential fatty
-
acid e.g. saf-
II PHOSPHOLIPID
ARACHIDONATE
Means and standard
Liver weight Phospholipid (mg/g liver) (9)
Treatment
E,@erimelzt
VALUES
error.
Phospholipid (weight %)
arachidonate (mglliuer)
(mglg liver)
I
--IS,
-EFI
7*
4.77
27.4 -h 1.1
13.7 & 0.4
12.8 & 0.8
2.7 f
-B, +B,
+EF_k +EFr\
7 7
4.65 4.30
28.6 * I.2 33.6 & 1.1
26.1 + 1.8 28.9 + 0.6
23.9 I 0.3 30.2 + 1.5
5.5 zt 0.6 7.0 i 0.3
-B, -B,
-EFh AEFA
7 7
2.99 3.26
30.4 & 0.8 29.7 i I.5
17.1 & 1.5 28.5 & 1.4
II.0 _k 0.9 20.0 t I.9
4.0 i 0.6 6.2 17 0.5
+B,
+EFA
7
3.72
30.1 =k 0.7
26.8 *
1.0
21.5 & 1.1
5.8 k 0.3
Expevime??l
0.2
2
-B, -B, +B,
-EFh +EFX +EFA
6 6 6
2.85 3.07 2.94
34.4 i 0.4 33.9 * I.2 38.0 & 0.6
15.2 & 0.9 26.0 f 1.1 30.4 & 0.6
10.7 & 0.7 19.3 + 1.2 24.4 * 1.0
3.8 * 6.3 i 8.3 :
-B, -BI +B,
-EFX +EF,\ --EFh
6 6 6
4.05 3.34 3.10
28.7 + 2.4 30.3 f 2.8 36.0 3 1.1
12.8 -& 0.6 28.4 * 0.9 32.2 * 1.2
10.7 + 0.9 20.4 * 1.5 25.9 i 1.5
2.6 f 0.2 6.1 + 0.3 8.3 & 0.2
Abbreviations: oil. * Number
B,, pyridoxine;
B,, riboflavin;
B,, thiamine;
EFA, essential fatty.acid
0.2 0.4 0.2
e.g. safflower
of rats per group.
After 6 days of linoleate suppementation, the rats were sacrificed by decapitation without anesthesia, Liver lipids were extracted, fractionated and fatty acids determined as previously described798. In the second experiment, the same dietary procedures were used, except that thiamine depletion, rather than riboflavin depletion, was produced. Pyridoxinedepleted
rats again were the controls. Biochim. Biophys. Acta, 137 (1967) 388-390
SHORT COhfMUh'ICATIONS
390
Liver phospholipid and phospholipid arachidonate values are given in Table II. In the first experiment, the degree of arachidonate depletion was similar in both vitamin-deficient groups at the start of the supplementation period (lines I and 4). In the group previously depleted of both pyridoxine and essential fatty acid, supplementation with pyridoxine and safflower oil produced a significantly higher level of arachidonate than did supplementation with safflower oil only.The difference in arachidonate was significant when expressed as mg/liver or as mg/g liver. The difference in weight o/0 was not significant. In the group previously depleted of riboflavin and essential fatty acid, supplementation with riboflavin and safflower oil, however, produced no greater increase in arachidonate than did supplementation with safflower oil alone. Therefore, riboflavin appeared to have no effect on arachidonate level. In the second experiment, the degree of arachidonate depletion was again similar in both vitamin-deficient groups at the start of supplementation (lines 7 and IO). In the group previously depleted of both pyridoxine and essential fatty acid, supplementation with pyridoxine and safflower oil produced a significantly higher level of arachidonate than did supplementation with the oil alone, in agreement with the first experiment. The differences were significantly greater when expressed as weight %, as mg/liver, or as mg/g liver. In the group depleted of thiamine and essential fatty acid, supplementation with thiamine and safflower oil also produced a higher level of arachidonate (weight o/O,mg/liver, mg/g liver) than did supplementation with safflower oil. Consequently, no evidence for a specific effect of pyridoxine in the conversion of linoleate to arachidonate can be inferred from this type of experiment in vivo since thiamine produced similar changes. Experiments ilz vitro may give more information on the role of pyridoxine or thiamine in arachidonate formation. GOSWAMI AND CONIGLIO~ recently observed a stimulatory effect of pyridoxal phosphate on the incorporation of [r-14]acetyl CoA into arachidonate and docosahexaenoate by liver microsomes. This study was aided by U.S. Public Health Service, Grant AM 7753. Department of Nutritional
Sciences
Ufiiversity of California Berkeley, Calif, (U.S.A.)
M.A. WILLIAMS D.J. MCINTOSH I. HINCENBERGS K.T.TAMAI
I J. F. MUELLER, T~itaminsHormones, 22 (1964) 787. 2 J. F. MEAD, Ann. Rev. Biochem.. 32 (1963) 241. 3 P. W. WITTEN AND R. T. HOLMAN, Arch. Biochem. Biophys., 41 (1952) 266. 4 J. C. KIRSCHMAN AND J. G. CONIGLIO, J.Biol. Chem., 236 (1961) 2200. =,G. E. SCHEIER AND M. A. WILLIAMS. Biochem. I.. 42 (146~1 422. 6 A. R. HANDS, N. S. SUTHERLAND A&W. BAR;L%,%&&~. J., 94 (1965) 279. 7 M. A. WILLIAMS, D. 1. MCINTOSH AND I. HINCENBERGS, 1. iVutvition, 88 (1966) rg-, s R. L. LYMAN, R. OS&ALD, P. BOUCHARD AND A. SHANNON, Biochem. J.,‘gS (;966) 4.38. g .4. GOSWAMI END J. G. CONIGLIO, Abstr., 7th Intern. Congr. Nutrition, (1966) 253.
Received
October
Biochim. Biophys.
rrth,
1966
Acta, 137 (1967) 388-390