By perfusion of isolated sections of intestine with a solution containing thiamine at a concentration of 3.1 micromole, it was established that thiamine absorption in animals fasted for 72 hours decreased by 28 percent, whereas absorption increased by 12 percent in rats after 24 hour immobilization. After immobilization, absorption of label in the intestinal mucosa increased. Na K ATPase activity in the intestinal mucosa decreased by 10 percent during fasting, and it increased with immobilization of the animals. Activity of Na K ATPase in the intestinal mucosa cells determined the absorption rate of thiamine and nicotinic acid at the level of vitamin transport through the plasma membranes of the enterocytes

Khmelevskiy, Y. V.

Kirilyuk, O. G.

English

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NASA TECHNICAL MEMORANDUM
	 NASA TM-76193
ABSORPTION OF THIAMINE AND NICOTINIC ACID IN THE RAC' INTESTINE
DURING FASTING AND IMMOBILIZATION STRESS
O.G. Kirilyuk and Yu.V. Khmelevskiy
(NASA-TM-76193) ABSORPTION JF THIAMINE ANDN ICOTINIC ACID IN THE SAT INTESTINE DURING
	
NEii7-3^J59
FASTING AND IMMOBILIZATION STRESS (NationalAeronautics and Space Administration) 11 piiC ADZ/MF A01
	
uucld zi
 06C 65/51 26666
Translation of "Vsmoktuvannya tiaminu to nikotynovoy kysloty
v kyshechnyku shchuriv pry holoduvanni to immobilizatsiynomu
stresi, Fiziolohichnyy Zhurnal, Vol. 23, No. 1, 1977,
pp 98-102
NATIONAL AERONAUTICS AND SPACE ADMINISTRATION
4iASHINGTON, D.C. 20546
	 JUNE 1980
STANDARD t1TLE PAGE
1. Rip:..t u=- 	 2. Government Accession No.
	 3. Recipiont's Catalog No.NASA TM-76193_	 I	 ,
d. Title and Subtitia ABSORPTION OF THIAMINE AN—D-i S. Pepart Date	 ^^
NICOTINIC'ACID IN THE RAT INTESTINE 	 June 1980
DURING FASTING AND IMMOBILIZATION	 6	 -- Performing Orgenisatton Cedernnn»n n	 {
7. Author(s) 0. G.	 Kirilyuk and Yu. V.
Khmelevskiy, Department of Biochemis-
try, Medical Institute, Kiev
1 8.	 Parr neing organisation Report No.
lo.	 Wa.k unit No.
11.	 Contract or Grant No.
.S` 3199
—^----- -
9. Performing Organi lotion Name and Address
13.	 Type of Repor t and Period CoveredLeo Kanner Associates
Redwood City, California 94063 Translation
12. Sponsoring Agency Name and Address
National Aeronautics and Space Adminis
tration, Washington, D.C. 20545 11
	 Sponsoring Agency Coda
15. Supplementary Notes
Translation of "Vsmoktuvannya tiaminu to nikoty-novoy kysloty
v kyshechnyku shchuriv pry holoduvanni to immobilizatsiynomu
stresi," Fiziolohichnyy Zhurnal, Vol.	 23, No. 1, 1977,
pp 98-102
16. Abstract
By perfusion of isolated sections of intestine with a
solution containing thiamine at a concentration of 3.1
umole, it was established that thiamine absorption in ani-
mals fasted for 72 hours decreased by 28% whereas ab-
sorption increased by 12% in rats after 24 hour immobiliza-
tion. Besides, after immobilization, absorption of label
In the intestinal mucosa increased. Na-K-ATPase activity
in the intestinal mucosa increased. Na-K-ATPase activity
in the intestinal mucosa decreased by 10% during fasting,
and it increased with immobilization of the animals. Evi-
dently, Na-K-ATPase activity in the intestinal mucosa cells
determined the absorption rate of thiamine and nicotinic
acid at the level of vitamin transport through the plasma
membranes of the enterocytes.
17. Key Words ( Selected by Author( s))
	
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ABSORPTION OF THIAMINE AND NICOTINIC ACID IN THE RAT INTESTINE
DURING FASTING AND IMMOBILIZATION STRESS
O.G. Kirilyuk and Yu.V. Khmelevskiy
Department of Biochemistry, Medical Institute, Kiev
One possible cause of endog-..ous vitamin deficiency is disturbance /98*
of the absorption of vitamins in the intestine [10, 201. Clinically,
the syndrome described is absorption deficiency, which is associated
with many diseases [1, 9]. However, the characteristics of vitamin
absorption in the intestine in pathologies, as well as the role of
separate processes which provide vitamin transport through the in-
testinal cell membranes, have been studied insufficiently.
We studied the characteristics of absor ption of thiamine and
i
nicotinic acid in the small intestine of
in animals under immobilization stress.
ATPase activity of the intestines of the
membrane enzyme, was studied [6, 13]. T1
of some substances through the membranes
the intestine [4, 12] .
the rat during fasting and
At the same time, ti).e Na +-K+
experimental animals, as a
zis provides active transport
of the epithelial cells of
Method
The studies were conducted on 50 white male rats, weighing
200+20 g, which were maintained on normal vivarium rations. The
characteristics of thiamine and nicotinic acid absorbtion were studied
under conditions of immobilization stress and fasting.
*Numbers in the margin indicate pagination in the foreign text.
1
The animals were immobilized for 24 hours in the horizontal posi-
tion, to prevent numerous ulcers in the stomachs of the animals. Upon
incision, the stomachs frequently are filled with blood, which is
characteristic of immobilization stress [17].
Rats of the other group were not fed for 72 hours (unlimited water)
before the experiment. After this time, the weight of the rats de-
creased by an average of 15%, but the weight of the intestine was un-
changed.
The absorption of thiamine in the rat intestine was studied, by
means of our modification of a method of perfusion of the intestine
[14]. Under hexenal anesthesia, the intestinal wall of the rat was
cut, and a section of the intestine 5 cm long was isolated, at the
level of the upper third of the small intestine, without damaging the
mesentery. The isolated section was perfused with a 0.9% sodium
chloride solution, which contained thiamine at a concentration of
3.1 pM. The perfusion rate was 0.75 mt/min. The perfusate was col-
lected fractionally every 2 min for a period of 10 min. Thiamine
absorption was determined from the decrease in thiamine in the per-
fusate. The thiamine was determined in the tests according to [3].
Nicotinic acid absorption was determined from the degree of dis-
appearance of the 14 C label, in administration of 14 C labelled nicotinic
acid to a 5 cm long section of the small intestine, isolated at the
level of the upper third of the small intestine. A 0.9% solution of
sodium chloride which contained 14 C nicotinic acid at a concentration
of 40 pM (activity of the preparation, 10 mC/g) was administered to the
section in the amount of 1 mt. After 10 and 60 min, the solution was
2
separated from the lumen of the intestine, and the degree of decrease
of the label	 in the contents of the section of intestine and the
amount of label in the mucosa were determined. The radioactivity of
the samples was determined in a PST-100 end counter. The results were
expressed in count/g/min.
The Na+-K+ ATPase activity was determined by the method proposed
by Govorova et al [2]. The results were expressed in pg
phosphorus/g tissue/hour.
The results were processed biometrically [5].
Results
Study of the absorption of thiamine by a section of the small
	
/99
intestine of the rat, during its perfusion for 10 min with a thiamine
solution (3.1 PM), permitted determination that, after 24 hour immobili-
zation of the animals, absorption of thiamine from the perfusate in-
creases by 12% (Fig. 1). While, in the control group of animals, of
the total amount of thiamine (7.5 pg), 4.25 pg of thiamine was absorb-
ed in 10 min, it was 5.25 pg in the rats which were immobilized. On
the contrary, in the animals which were fasted for 72 hours, thiamine
absorption decreased by 28% and, of the total amount of thiamine (7.5 pg),
only 2.25 pg was absorbed (Fig. 1).
The change in the thiamine absorption rate, which was determined
at 2 min intervals over 10 min, is in the same direction as that of the
data presented (Fig. 2). In immobilization stress, the thiamine ab-
	 /100
sorption rate increases to 0.55 pg/min, and it decreases to 0.23 pg/min
in the animals which were fasted, with a standard 0.45 p g/min of thia-
mine in the perfused section of the small intestine.
3
aFig. 1. Absorption
of thiamine in rat in-
testine (in % of total
amount in perfusate):
1. control animals; 2.
animals under immobili-
zation stress; 3. animals
fasted for 72 hours.
Key: a. min
a
b
Fig. 2. Rate of absorp-
tion of thiamine in rat
intestine (in u g thiamine
per min): conventional
designations as in Fig. 1.
Key: a. ug	 b. min
We determined nicotinic acid ab-
sorption in the small intestine of normal
rats and in animals which endured immobili-
zation stress. The study results are
presented in Fig. 3. They characterize the
degree of decrease of the 14C label in
the luminal contents, 10 and 60 min after
administration of the 14C nicotinic acid
solution to the lumen of the intestine.
A higher degree of absorption of nicotinic
acid from the lumen of the intestine of the
rat during immobilization stress was
found. The difference in the decrease in
label was a little higher in 10 than in
60 min.
Fig. 4 presents the results of study
of the accumulation of the 14C label of
nicotinic acid in the mucosa of the sec-
tion of intestine of normal rats and
those which endured immobilization stress.
A sharp increase in accumulation of label
in 10 min was traced in the intestinal mucosa of the rats which endured
immobilization. Besides, the difference in accumulation of label in
the mucosa was less pronounced in 60 min, although the direction of
the proc6ss is the same.
Study of Na + -K+ ATPase showed that the activity of the enzyme in
the intestinal mucosa of the rat depends on the condition of the body
4
ba
Fig. 3. Nicotinic
acid absorption
from rat intestinal
lumen: conventional
designations as in
Fig. 1.
Key: a. count/min/mi
b. min
of the animal. Fig. 5 presents the results
of determination of Na + -K+ ATPase activity
in the intestinal mucosa of the rat under
immobilization stress and in fasted animals,
compared with normal rats. In 24 hour
immobilization, the activity of the enzyme
under study increased sharply, and it is
nearly 50% above the Na + -K+ ATPase activity
of the mucosa of the small intestine of
normal animals. In 72 hour fasting, a
tendency develops toward a decrease in
activity of the enzyme, compared with the
group of control animals.
Discussion
It was determined from the study that,
under immobilization stress conditions,
thiamine and nicotinic acid absorption ir
the rat intestine increases. At the same
time, Na+ -K+ ATPase activity in the mucosa
of the upper third of the small intestine of
the rat increases.
Fig. 4. Nicotinic
acid accumulation in
rat intestinal mucosa
upon absorption from
intestinal lumen: ^,!c>n-
vent tonal dens l gna ' io r..--
as in Fig. l..
Key: a. count/min/g
b. min
According to the data of Tursunkhodz-
hayeva et al C31, the consumption of group B
VI '6,	 increases under stress. It follows
from the data we obtained that the increase
of vitamin consumption is produced by increase
5
in absorption in the intestine, due to an increase
In activity of Na+ -K+ ATpase, which participates
in transmembrang transport of matter [4].
It is known that Na +-K+ ATPase is a membrane
fragment f5], localized chiefly in the lateral and
basal plasma membranes of the enterocytes [7, 131.
The entry of thiamine into the blood and lymph ap-
parently is connected with the functioning of this
enzyme [181. It is known that the activity of this
enzyme is under hormonal control, since the possi-
J.
ro
e
t
2
0
9	 J
Fig. 5 . Na'' -K+
ATpase activity
in mucosa of rat
small intestine:
conventional
designations as
in Fig. 1.
Key: a. mg
bility of activation of Na+-K+ ATPase in some tissues of animals by
corticosteroids [131, the content of which increases under stress [111,
has been established.
The decrease J_.n thiamine absorption in fasting animals must be
explained by decreases in Na + -K+ ATPase activity in the intestinal
mucosa. It must be noted that some authors point out a decrease in
activity of the intestinal enzymes which participate in digestion and
absorption, when peroral intake of foodstuffs to the body is stopped
[151, or in parenteral feeding [161, which is accompanied by a decrease /101
in the absorptive capacity of the intestine.
Therefore, the absorption of thiamine and nicotinic acid in the
rat intestine in small physiological concentrations (3.1 and 40 pM) de-
pends on the activity of membrane Na + -K+ ATPase, which provides active
transport through the plasma membrane of the cells of the intestine
[12]. A reduction in activity of this enzyme by hormones or other
actions (anorexia, exclusion of the functional activity of the intestina)
6
affects the level of absorption. of the coenzyme vitamins thiariine and
nicotinic acid in the intestine which, in the final analysis, can
affect the provision of the body with these vitamins.
dd
(^Opl 4/69r.10 V Yt AVIV Ts Tvo
to Lok k cI ov
f(	 I/
2 1. ^,^ ^^o ^,19-77.
REFERENCES
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ATPase activity of the brain and erythrocytes in anoxia,"
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3. Yeliseyeva, G.D., "Fluorometric determination of thiaming, co-
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8
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9


Absorption of thiamine and nicotinic acid in the rat intestine during fasting and immobilization stress

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