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5MEDICAL SCIENCES
INFLUENCE OF CHRONIC TOXIC HEPATITIS ON THE COMPOUND OF MILK IN THE
DYNAMICS OF LACTATION
KhasanovB.
Bukhara State Medical Institute DOI: 10.5281/zenodo.6806972
Abstract
The effect of chronic toxic hepatitis (CTH) of female rats on the compound of milk in the dynamics of lactation was researched on mature females. It has been established that autoimmune processes are activated in the body of females with CTG during breastfeeding, and antihepatocyte antibodies are detected in blood serum and milk. Significant amounts of autoantibodies in the dynamics of lactation are determined mainly in the blood serum, while in breast milk they are insignificant. However, a pronounced decrease in the number of immunocompetent milk cells in the dynamics of lactation in females with CTG most likely indicates a violation of the transfer of adoptive immunity and the immunomodulatory function of mother's milk.
Keywords: hepatitis, antihepatocyte antibodies, lactation, blood serum, milk, immunocompetent cells.
Introduction. The frequency of various extra genital pathologies in women of childbearing age is still significant. First, the frequency of chronic damage to the hepatobiliary system increases because of viral or toxic hepatitis, which, under the influence of adverse environmental conditions, often take a chronic, protracted form [1, 4, 9].
It has also been established that the mammary glands after childbirth are the only organ that connects the body of the mother and the baby. The transfer of adoptive immunity, as well as many biologically active substances through mother's milk, is decisive for the formation of the immune, digestive and many other systems of the newborn's body [2, 7, 10, 17, 22, 23, 27, 29, 30]. However, a violation of this harmonious system occurs with extra genital pathology of the mother, including hepatitis.
The possibility of transmission of the hepatitis virus from women carriers of HbsAg to children born, the development of HbsAg persistence in some children, the formation of primary chronic hepatitis is beyond doubt [13, 20, 26]. At the same time, the significance of autoimmune processes that unfold in the mother's body in chronic toxic (CHT) hepatitis, in the dynamics of lactation and their influence on the qualitative composition of breast milk and the development of offspring during breastfeeding, still remains poorly understood. Since there are a significant number of researches indicating that in newborns from mothers with hepatic pathology, one of the most common symptoms is: a violation of the digestive processes [5, 12], a slowdown in the formation of the immune components of the small intestine, as well as the organs of the immune system of the offspring in the early postnatal period of ontogenesis [4, 6, 8, 16, 18, 19, 20, 24]. However, the question of the causes and mechanisms leading to these changes remains open; perhaps it may also consist in a change in the qualitative composition of mother's milk.
Thereby, the purpose of our research was to stud<y the effect of experimental mother's CTG on the qualitative composition of mother's milk in the dynamics of lactation.
Material and research methods
For the experiment, 3-month-old mature white outbred female rats (72) weighing 120-140 grams were used. The animals were kept on the usual laboratory diet and were kept in quarantine for two weeks prior to the experiment. As a model of hepatitis, we used chronic heliothrine intoxication [1]. After the quarantine period, the females of the experimental group (O) were injected with heliothrin at a dose of 0.05 mg/gram of body weight per 0.5 ml of saline subcutaneously weekly for 6 weeks, the animals of the control group (K) were injected with only saline. Males were added to the females 10 days after the last injection. For the study, females of the experimental (40) and control groups (32) were selected on days 1, 3, 7, 15, 21 and 30 after birth. Determination of antihepatocyte antibodies was carried out by RPHA according to the Boyden method. To conduct serological reactions, blood serum (0.5 ml) and milk samples (0.2 ml) taken from female rats in the above periods of the study were used. Sheep erythrocytes were prepared according to the generally accepted method. As an antigen, a sample of the liver of an intact female rat for 2.0 g was used, the preparation of which was carried out by water-chloroform extraction according to the method of T.A. Alekseeva [14]. The content of the antigen was controlled by the amount of protein, which was adjusted to 1%. For more accurate calculations and convenience in comparison, the results obtained were expressed in log2 [11]. Additionally, the most accessible method for assessing the immunomodulatory function of the mammary gland, we decided to use immunocompetent milk cytometry in the dynamics of lactation. For cytometric researches of milk cells, smears were prepared and stained according to May-Grunwald. ICCL counting was performed in 20 fields of view from each preparation (Approx. 15 x 90) where five preparations from each animal were researched. The data obtained were processed according to Fisher-Styudent, differences were considered significant if they satisfied P<0.05.
Results
When researching the possibility of autoimmune processes in maternal CTG, we established a certain dy-
namics of changes in the titer of antihepatocyte antibodies in the blood and milk of rats. In the blood serum of the females of the experimental group, immediately after childbirth, autoantibodies reaching a titer of 1:128 are detected, this trend persists up to 3 days of lactation, while in the control group of animals, and the titer of autoantibodies is 1:8 and 1:4 on days 1 and 3 after childbirth, respectively. In subsequent periods of lactation, sera at a dilution of 1:64 turned out to be seropositive until the end of the lactation period. On the contrary, in the control group, antibodies are found in females only up to 7 days of breastfeeding, not exceeding a titer of 1:4, only traces were observed in the subsequent periods of lactation.
In contrast to blood serum, in a serological research of milk samples from females of the experimental group on the 1st day after birth, autoantibodies
were determined at a dilution of 1:8. In subsequent periods, anti-hepatic antibodies were not always detected even in a 1:4 dilution. In the control group, the indicators of serological studies of milk during breastfeeding did not find significant differences from the experimental group. For more accurate studies and the possibility of comparing changes in the titer of antihepato-cyte antibodies in the blood serum and milk of female rats in the dynamics of lactation, the results of serolog-ical studies were expressed in log2 (see Table 1). According to these data, in toxic heliotrine hepatitis, the appearance of autoantibodies against the liver is characteristic only for the blood serum, which are present until the end of the breastfeeding period. At the same time, in milk samples obtained from females with toxic hepatitis, the titer of autoantibodies did not differ significantly from those in females of the control group.
Table 1.
Changes in the activity of anti-hepatic antibodies in blood and milk Female rats with chronic heliothrine intoxication in the dynamics of lactation
Researched material Group of animals Terms of lactation (days)
1 3 7 14 21
Blood C 1,68±0,158 0,75±0,161 0,68±0,149 0,48±0,142 0,33±0,104
E 6,03±0,089 5,45±0,127 5,10±0,114 4,90±0,120 4,73±0,114
Milk C 1,60±0,206 1,48±0,089 1,30±0,082 0,83±0,196 0,78±0,164
E 3,75±0,133 3,45±0,158 3,43±0,212 2,00±0,199 1,95±0,225
Note: * - differences are significant relative to control at P<0.05
Cytometric studies of milk ICCL showed that the main type of ICCl supplied with breast milk to rat pups are monocytes and macrophages, as well as small lymphocytes (Table 2). In chronic heliothrine intoxication, from the first days of lactation, a decrease in the total number of ICCls is observed, their most pronounced decrease occurs on the 15 th day of lactation. It should also be noted that by the end of the lactation period, in contrast to the control group of animals, the milk of females with toxic hepatitis does not contain ICCL.
Table 2
Quantitative relationship of immunocompetent cells of milk of female rats with heliotrine hepatitis in the dynamics of lactation _(Absolute values. X ± xm, with n=8)_
Terms of lactation Group Immune Component Cells
of animals Total number Big lymphocytes Middle lymphocytes Small lymphocytes Monocytes Macrophages
C 216,3± 11,1± 21,0± 28,8± 109,1± 46,3±
1 day 15.0 0,8 1,6 1,2 6,0 2,60
E 139,0± 3,1± 12,6± 18,5± 72,6± 32,1±
11,4 0,39 0,52 1,68 2,56 1,81
C 175,3± 6,0± 22,6± 39,8± 75,8± 31,1±
3 days 6,60 0,71 2,03 1,48 3,87 2,34
E 126,3± 3,5± 15,3± 24,5± 64,8± 18,3±
10,41 0,35 1,13 1,94 4,22 1,75
C 122,1± 5,0± 16,9± 15,8± 52,3± 32,3±
7 days 9,23 0,53 0,66 0,86 2,72 2,01
E 77,4± 2,8± 10,1± 10,3± 36,8± 17,5±
6,80 0,42 1,37 1,22 2,28 1,06
C 45,1± 2,8± 14,1± 7,3± 11,3± 9,8±
15 days 2,78 0,24 1,01 0,68 0,33 0,68
E 21,6± 1,8± 5,6± 3,5± 7,9± 2,9±
1,94 0,24 0,35 0,27 0,39 0,39
C 8,5± 1,0± 1,4± 1,8± 2,1± 2,3±
21 days 0,44 0,35 0,44 0,33 0,39 0,24
E 0 0 0 0 0 0
Note: 1. Symbols: Groups of animals. C - Control. E-experienced.
2. * - indicators where the differences are significant relative to the control at P<0.05 are in bold.
Discussion
Analyzing the obtained results, it should be noted that chronic heliothrine intoxication leads to the occurrence of toxic hepatitis, which generally tends to progress, i.e. is a model of aggressive chronic hepatitis [1, 3]. In active forms of hepatitis, changes in the immune status occur, in particular, there are profound changes in the T- and B-systems of immunity and there is an increase in the titer of immunoglobulins of various classes, an increase in the gamma fraction of globulins, which is the criteria for assessing the activity of the developing pathological process. In cases where the process becomes chronic, against the background of a slight decrease in T-helpers, the number of T-suppres-sors (killers) is significantly reduced, which contributes to the formation of anti-hepatic antibodies and activation of the pathological process with the inclusion of an autoimmune mechanism [3, 9, 23, 24].
The results of our research showed once again that heliothrine intoxication in female rats before pregnancy is a trigger mechanism for an autoallergic process that progresses over time. Along with this, relatively low titers of anti-hepatic antibodies in the blood serum, apparently, are associated with changes occurring in the mother's body in the dynamics of pregnancy and lactation, stimulating regeneration processes. The almost twofold excess of the autoantibody titer immediately after childbirth, relative to the further period of lactation (1:128 on the 1st day and 1:64 in the subsequent periods of lactation), may be due to the effect of the fetus, foreign to the mother. The presence of a small amount of antibodies in milk during lactation indicates that the cause of dystrophic changes in the liver of rat pups born to intact rats, when fed by a female with he-liotrine hepatitis [4], are not autoantibodies, but possibly hepatotoxins formed in the mother's body , due to liver dysfunction [10, 11, 12]. If we also take into account the decrease in ICCL in females with CTH in the dynamics of lactation, which enters the body of the newborn with mother's milk, apparently, is one of the factors characterizing the decrease in the immunomod-ulatory function of milk. In addition, a decrease in the number of macrophages, monocytes and lymphocytes, on the one hand, contributes to the disruption of the transfer of adoptive immunity, on the other hand, as was found in our previous studies [15, 30], the supply of lysosomes, lipid droplets present in these cells and the trophic influence and immunobiological properties of breast milk are significantly reduced, allowing the child to adapt and survive in the "world of microbes", which he enters immediately after birth [21, 25, 28].
Thus, our results allow us to draw the following conclusions:
1. In the blood of female rats with toxic hepatitis after childbirth and in the dynamics of lactation, anti-hepatic antibodies are determined, but during breastfeeding, they are transmitted to rat pups through milk in small quantities and, most likely, are not the main reason for the lag in the development of the organs of the digestive and immune systems of the offspring.
2. The decrease in the amount of ICCL of milk found in females with CTG from the first days after
birth and in the dynamics of lactation and the characteristic absence of ICCl in the milk of hepatitis females on the 21st day of lactation indicates a violation of the transfer of adoptive immunity to offspring, i.e. im-munomodulatory function of mother's milk, so necessary for offspring in the period of early postnatal ontogenesis.
References
1. Abdullaev N.Kh., Karimov Kh.Ya. Liver in case of intoxication with hepatotropic poisons. - T.: Medicine. -1989. - 140 s.
2. Avlod S., U. B., Developing (2020). Artificial feeding and features of the development of offspring, and the formation of the adrenal glands in early postnatal ontogenesis.
3. Azizov E.Kh. Structural and functional bases of the reaction of mesenteric lymph nodes in experimental toxic chronic hepatitis and ways of its correction: Abstract of the thesis. Candidate of medical sciences: 14.00.23. - Tashkent, 1996. - 19 p.: ill.
4. Azizova F. Kh. & Otajonova A. N. (2010). Structural features of the formation of Peyer's patches in offspring under conditions of chronic toxic effects on the mother's body. Morphology, 117(4), 13-14.
5. Azizova F.Kh., Bajakova D. B., & Akhmedova H. Yu. (2001). Age-related structural and functional features of the small intestine of rat pups born from female rats with chronic toxic hepatitis. Doctor. Case, 1, 103.
6. Azimova S.B. et al. (2021). Toxic hepatitis of the mother and structural and functional formation of the offspring thymus in the dynamics of early postnatal ontogenesis. Eurasian Journal of Academic Research, 1(9), 426-429.
7. Influence of breast milk enzymes on the metabolism of carbohydrates in the process of adaptation of newborns in the early neonatal period / Nabuhotny T.K. Markevich V.E., Pavlyuk V.P., Tkachenko Yu.P. // Nutrition issues. - 1985. - No. 3. - C. 34-37.
8. Gafarova E.A. Initial and final stages of carbohydrate hydrolysis in the offspring of rats with chronic toxic hepatitis: Abstract of the thesis of a candidate of medical sciences. - Tashkent. 2001. - 18 p.
9. Eremina E.U. Autoimmune liver disease and pregnancy. // Practical medicine. - 2011. - (54). - S. 1218.
10. Zufarov K.A. and other Quantitative and ultrastructural characteristics of immunocompetent cells of the mammary gland in the dynamics of pregnancy and lactation // Morphology. - 2003. - Volume 124, No. 4. - S. 74-79.
11. Instructive and methodological materials on the use of serological diagnostic methods in epizooto-logical examination of natural plague foci. - Moscow. Ministry of health of the USSR. GOOKS. - 1983. - 135 p.
12. Kurbanov A.Sh. The influence of protein deficiency in the mother's diet during pregnancy and lactation on the development of physiological systems of the initial and final stages of carbohydrate hydrolysis in the offspring: Abstract of the thesis of candidate of biological sciences. - Tashkent. 1997. - 24 p.
13. Minoranskaya E.I. Carriage of HBsAp in pregnant women: clinical and biochemical characteristics and impact on pregnancy and offspring: Abstract of the thesis candidate of medical sciences. L., 1990. - 19 p.
14. Nikolaev A.I., Platonova L.E. Methods for the determination of antibodies and their comparative evaluation. - T., "Medicine". - 1971. - 112 p.
15. Sultanova D.B., & Khasanov, B. B. (2019). Influence of toxic hepatitis at period of the lactations on haematologic indexis mother's and posterity. A new day in medicine, (4), 419-421.
16. Features of the formation of the organs of the immune system in rat pups born from females with chronic toxic hepatitis / Azizova F.Kh., Tukhvatulin F.F., Tursunmetov I.R., Tukhtaev K.R. // Medical practice (Kiev "Health"). - 2001. - No. 2. - P.100-103.
17. Khasanov B.B. Gamogenesis endocrine management // New Day in Medicine. - 2019. - No. 4. - 92100 b.
18. Khasanov B.B. Hormonal components of breast milk and their physiological significance. Problems of biology and medicine - 2021, 3 (128), 271-276.
19. Khasanov B. B., & Sultanova D. B. (2020). The influence of extragenital pathology of the mother on the postnatal development of the liver and kidneys of the offspring. In University Science: Looking to the Future (pp. 657-659).
20. Khasanov B. B., & Khasanova Z. Sh. Effect of toxic hepatitis on the reproductive function of female rats and the development of the offspring immune system. Problem of biology and medicine -2003, 2, 65-69.
21. Eberl G. A new vision of immunity: homeostasis of the superorganism. Mucosal Immunology. Vol. 3 N 5. September 2010. doi: 10.1038/mi.2010.20 - P 450-460.
22. Hassiotou F., Geddes D.T. Immune cell-mediated protection of the mammary gland and the infant
during breastfeeding // Adv. Nutr. 2015. V. six, No. 3. P. 267-275.
23. Khasanov B. B. (2020). Experimental chronic toxic hepatitis and hematological features in the dynamics of mother's and the offspring lactation. European Journal of Molecular & Clinical Medicine, 7(09), 1367-1373.
24. Khasanov B. B. (2021). Structural and functional features of immunocompetent breast cells glands during pregnancy and lactation in chronic hepatitis. Psychology and Education, 58(2), 8038-8045.
25. Parker M. G., Stellwagen L. M., Noble, L., Kim, J. H., Poindexter B. B., & Puopolo K. M. (2021). Promoting human milk and breastfeeding for the very low birth weight infant. Pediatrics, 148(5).
26. Protecting the Newborn and Young Infant from Infectious Diseases: Lessons from Immune Ontogeny. Immunity Review. Immunity 46, March 21, 2017 a 2017 Elsevier Inc. P/ 350-363.
27. Sultanova D. (2021). The influence of mother's extragenital pathology on the formation spleens of offspring in the early period postnatal ontogenesis. The Scientific Heritage, (81-2), 47-49.
28. Taranushenko T.E. Unity of bowel-lung axis and the role of beneficial microbiota in anti-infectious protection. Review Articles. Russian Journal of Woman and Child Health. 2021; 4(4):355-361 (in Russ.). DOI: 10.32364/2618-8430-2021-4-4-355-361.
29. Tukhtaev K.R. and others. (2003). Structural and functional interrelations of immunocompetent cells in the mammary gland of lactating rats and in the small intestine of newborn rats during suckling period. Morphology (Saint Petersburg, Russia), 124(6), 70-72.1
30. Zufarov K.A. and others. (2003). Quantitative and ultrastructural characteristics of immunocompetent cells in the mammary gland during pregnancy and lactation. Morphology (Saint Petersburg, Russia), 124(4), 74-79.
