CC BY
7EFFECT OF CHRONIC TOXIC HEPATITIS OF THE MOTHER'S FORMATION OF THE SPLEEN OF THE OFFSPRING DURING THE BREASTFEEDING PERIOD
KhasanovB.
Department, of Histology, cytology and embryology
Sultonova D.
Department, of Biological Chemistry Bukhara State Medical Institute, Bukhara, Uzbekistan DOI: 10.5281/zenodo.6695594
Abstract
It has been established that chronic toxic hepatitis is one of the common diseases of the gastrointestinal tract, including the cause of extragenital pathology in women of childbearing age. The effect of chronic toxic hepatitis in female rats on the growth and development of the offspring spleen during early postnatal ontogenesis was researched. It has been established that chronic maternal hepatitis leads to a pronounced lag in the development of the spleen during the period of pure breastfeeding, the transition of rat pups to mixed nutrition had a positive effect on the development of the structural and functional zones of the spleen.
Keywords: early postnatal ontogenesis, maternal hepatitis, development, spleen, rat pups.
It is known that the spleen is not one of the vital organs; however, it is the largest collector of lymphoid tissue in the body and performs important hematologi-cal and immunological functions, forming a generalized immune response of the whole organism to antigen penetration, inflammation, or any other violation of the body's homeostasis [9]. In the spleen, an active and very long-term contact of variously determined immu-nologically competent cells with antigens in the blood passing through the spleen is provided [15, 17].
Along with this, it has been established that the spleen is the largest lymphoid organ in the human and mammalian body, making a significant contribution to the development and maintenance of the cellular and humoral immune response, innate and acquired immunity, the quantitative and qualitative composition of blood immunocytes, lymph and other lymphoid organs [16 , eighteen]. The interaction and establishment of links accompany its development in early postnatal ontogenesis between hematopoietic and stromal cell populations of numerous micro anatomical zones on the territory of the organ. The lymph-destructive, lymph-degenerative, and lymphoproliferative effects of stress can change these relationships and lead to a long-term impairment of the immune response [4, 5]. However, a complete understanding of the nature of the stromal cells of the spleen, their origin, patterns of distribution over sub compartments of the organ and mechanisms of influence on lymphoid cells under stress has not yet been achieved; the nature of the follicular dendritic cells has not been determined; regularities of maturation and differentiation of dendritic interdigitating cells have not been elucidated; there are disagreements in understanding the nature of reticular stellate cells, etc., which is associated with a lack of research on the lym-phoid and stromal architectonics of immunogens organs, especially in childhood, and makes it difficult to understand the patterns of formation of an effective immune response in early postnatal ontogenesis [7].
Despite ongoing therapeutic and preventive measures, chronic hepatitis is still one of the most common diseases, and often the cause of extra genital pathology, among women of childbearing age [6, 24].
Naturally, a violation of the functional state of the liver during pregnancy and breastfeeding has a negative impact not only on the mother's body, but also on the developing offspring [2, 22, and 23]. At the same time, in the literature available to us, we were not able to find works devoted to the research of the influence of a chronic mother on the formation of the spleen of the offspring during breastfeeding. In this regard, the purpose of our work was to study the effect of experimental toxic maternal hepatitis on the structural and functional development of the offspring spleen during early postnatal ontogenesis.
Material and methods The experiments were carried out on 60 mature female white outbred rats weighing 130-150 g. All animals were kept under quarantine for two weeks to exclude infectious diseases. Experimental animals were divided into 2 groups (30 rats in each): experimental and control. A model of chronic toxic hepatitis (CTH) was obtained using heliothrin according to the generally accepted method [1], by weekly administration of 0.05 mg of heliothrin per 1 g of body weight for 6 weeks. The control group of females received sterile saline instead of heliothrin. Males were added to the females 10 days after the last injection. Subsequently, the course of pregnancy was monitored. Rats were decapitated under light ether anesthesia on days 1, 3, 7, 15, 21, and 30 after birth. Pieces of the spleen were fixed in 12% neutral formalin and Bouin's fluid, and after appropriate wiring, they were embedded in paraffin. Sections 5-7 ^m thick were stained with hematoxylin and eosin and used for morphological and morphometric researches. Statistical processing of the obtained data was carried out using computer programs.
Results and discussion Because of our research, it was established that the spleen of rats undergoes certain structural and functional rearrangements in the process of postnatal growth. In newborns, during the first day after birth, the spleen parenchyma is mainly represented by red pulp. Outside, the organ is covered with a thin connective tissue capsule, from which delicate layers of connective
tissue extend into the organ. During this period of postnatal life, the spleen of rats is characterized by the presence of many erythroid cells at various stages of differentiation, diffusely distributed lymphoid cells, granulo-cytes and megakaryocytes, as well as the absence of a formed white pulp. On the 3rd day of postnatal development, it is characterized by the relative development of the cellular components of the red pulp, pronounced processes of thrombopoiesis and erythropoiesis, the appearance of muscle-type arteries and the beginning of the formation of the lymphoid apparatus of the spleen. Significant restructuring of the stroma and parenchyma of the spleen characterize the 7th day of postnatal development. During this period, the formation of the reticular base of the organ, the pronounced development of the vascular system and the differentiation of the red and white pulps take place. On the 15th day, further formation of the stroma and parenchyma of the spleen is observed. This period is characterized by a decrease in the rate of erythropoiesis and thrombopoiesis in the spleen, and, on the contrary, intensification of the processes of lymphocytopoiesis and the formation of the white pulp of the organ. On the 21st day of postnatal
ontogenesis, the predominant development of the white pulp of the organ is observed. In this case, there is an increase in the number and expansion of the diameter of the lymph nodes. It should be noted that in the lymph nodes. As in the previous periods of the study, germinal or light centers are not clearly expressed. At the same time, in each nodule, 3 zones can be distinguished during this period. Along with the periarterial and marginal zones, an intermediate zone is identified during this period, located between these zones. Thus, on the 21st day of postnatal development, the formation of a white pulp occurs, where the processes of lymphocyto-, plasmacytopoiesis and antibody formation take place. At the same time, the intensity of erythropoiesis and thrombopoiesis significantly decreases. An increase in the number of macrophages in the red pulp probably indicates that in this period the spleen begins to actively function as a hematopoietic organ. Further growth of the organ is accompanied by its quantitative and qualitative changes. By the 30th day of postnatal development, the spleen acquires a structure characteristic of the spleen of adult animals.
Table 1
Influence of maternal toxic hepatitis on some anthropometric parameters and spleen mass in rat pups in the dy-__ namics of early postnatal ontogenesis (X±Sx)_
Researched parameters Group of animals Age of animals (in days)
1 3 7 15 21 30
Rat weight, g C 5,6±0,08 6,0±0,09 8,9±0,11 18,6±0,43 29,1±0,82 42,3±0,75
E 5,1±0,11 5,3±0,13 8,1±0,12 16,9±0,42 26,8±0,55 34,7±1,14
Rat body length, mm C 27,1±0,47 33,0±0,58 44,2±0,39 51,6±0,52 55,2±0,88 57,6±0,96
E 23,9±0,43 27,1±0,52 36,4±0,48 41,5±0,52 47,6±0,82 51,6±1,05
Absolute weight of the spleen, g C 7,4±0,34 20,9±0,89 31,6±1,04 67,7±2,92 78,6±3,58 138,1±4,84
E 8,3±0,36 16,0±0,45 22,9±0,69 71,9±3,12 119,6±4,13 181,6±4,25
Note: 1. conditional abbreviations, group of animals, C - control, E-experienced;
2. Bold type indicates the values where the differences are significant relative to the previous period at
P<0.05.
Table 2
Average number and diameter of splenic lymph nodes rats in the dynamics of early postnatal ontogenesis
№ Researched parameters Group of animals Rat age in days
7 15 21 30
1 Average number of nodules C 5,8±0,10 7,3±0,22 8,7±0,31 12,5±0,31
E 4,7±0,11 6,2±0,13 7,6±0,24 12,3±0,25
2 Average number of diameter C 176,4±1,6 278,5±3,3 329,6±2,5 338,2±3,2
E 160,4±1,8 261,3±2,6 322,3±3,2 335,8±2,9
As shown by the results of our research presented in the table, maternal CTG contributes to growth retardation in offspring, expressed in a decrease in body weight on days 1-3 after birth and body length of animals, noted on days 7-14 after birth. Also, a decrease up to 7 days, a decrease in weight, and then, on days 15-21 after birth, an increase in the weight of the spleen of rat pups was also established (see Table 1). In addition, when studying the morphometric parameters of the spleen of rat pups of the experimental group, it was found that the average number and average diameter of lymphoid nodules decreased up to 15 days of postnatal ontogenesis (see Table 2).
A completely different picture was observed in morphological researches of the spleen of rat pups born from mothers with CTG, where a significant delay in the postnatal formation of structural and functional zones was noted. On the 7-14th day of the experiments, the parenchyma of the lymphoid follicle consisted mainly of diffuse lymphoid tissue without clear lines between the red and white pulp of the organ. The sub-capsular sinuses were significantly dilated. The absence of formed lymphatic follicles during this period is characteristic of the spleen and, as established by other researchers, for lymph nodes and Peyre's patches [3, 10, 12, 25, and 26]. On the 14th day in the rat pups
of the experimental group, only the formation of small lymphoid follicles was noted, but their number and diameters were significantly smaller and lower than those in the spleen of the control group of rat pups. In the control group, sufficiently formed lymphatic follicles were detected during this period. Even by the 21st day of the postnatal period, in the rat pups of the experimental group, the lymphoid tissue of the spleen remained hypoplastic, without clear boundaries of the various structural zones of the lymphatic follicles.
Analyzing the obtained results, it is necessary to point out that man, like many mammalian species, brings immature offspring. To compensate for this im-munological immaturity inherent in the period of the fetus and newborn, as well as the first months of life, nature has developed adaptive defense mechanisms provided by the mother, represented by the transplacen-tal transfer of antibodies, anti-infection resistance factors in the amniotic fluid, and after birth, in colostrum and milk. Therefore, for the full development of the baby, starting from the first day after birth, protein components are needed: "the main plastic material", and, of course, easily digestible carbohydrates of mother's milk. In addition, a certain dynamics of the level of hormones in milk is revealed, associated with their participation in the process of metabolic adaptation of new-borns to extrauterine existence and causing the restructuring of protein, carbohydrate and fat metabolism in the postnatal period. The enzymes of colostrum and milk that enter the body of a newborn during breastfeeding also have a beneficial effect on the processes of its adaptation, affecting the metabolism of proteins and carbohydrates in the intestine [8, 11, 13, 14, 19, 20, 26, and 28]. It has been established that in the offspring of rats with chronic hepatitis, there is a steady decrease in body weight gain, and a lag in the structural and functional development of the small intestine [10, 21, and 23]. Perhaps one of the reasons contributing to these changes during the period of lactotrophic nutrition is the previously discovered decrease in the amount of protein, carbohydrates and enzymatic activity of milk. The decrease in cellular components, apparently, is one of the factors characterizing the decrease in the im-munomodulatory function of milk. In addition, a decrease in the number of macrophages, monocytes and lymphocytes, contributing, on the one hand, to the disruption of the transfer of adoptive immunity. In addition, as indicated in our previous studies, the supply of lysosomes, lipid droplets present in these cells, is significantly reduced.
It is known that the postnatal formation of lym-phoid tissue depends on many internal and external factors, among which the antigens of the external environment play an exceptional role. With exogenous anti-genic exposure, a significant acceleration of the formation of lymphatic follicles and differentiation of their structural and functional zones was noted [7, 16, and 27]. However, the natural formation of lymphoid tissue under the influence of environmental antigens can occur only if the processes of embryonic laying and intrauterine development of the immune system organs are normal. Undoubtedly, chronic hepatitis, accompanied by a number of metabolic disorders in the body,
changes the natural course of the embryonic development of organs and systems, including the immune system, and in particular the spleen. These disorders continue to manifest themselves in the postnatal period of life. Our data indicate a significant delay in the postnatal formation of the spleen system in toxic hepatitis in the mother.
Thus, because of our research, it was found that in rat pups born from females with chronic heliotrine hepatitis, there is a lag in the structural and functional formation of the spleen in the initial period of breastfeeding, the transition of rats to mixed feeding has a positive effect on the development of the spleen. Therefore, therapeutic and preventive measures for offspring born from mothers with toxic hepatitis would be more appropriate, starting from the antenatal period and continuing during breastfeeding.
References
1. Abdullaev N.Kh. Liver in case of intoxication with hepatotropic poisons. / N.Kh. Abdullaev, Kh.Ya. Karimov. - Tashkent.: Medicine. - 1989. - 140 p.
2. AVLOD S., USTI V.U.B., (2020). Artificial feeding and features of the development of offspring, and the formation of the adrenal glands in early postnatal ontogenesis.
3. Azizova F.X., A.N.O., Ishandjanova S.X., Umarova Z.X., & Utepova N. B. (2021). Morphological characteristics of T-dependent zones of the organs of the immune system in chronic intoxications.
4. Afanaseva M. A. Development of the immune system in the ontogeny of rats: neuroendocrine-im-mune interactions: author sciences: spec. 03.00.17 "Physiology" / M. A. Afanaseva. - M., 2009. - 22 p.
5. Bryukhin G.V., Fedosov A.A. Characterization of the proliferative activity of thymocytes and peripheral blood lymphocytes in the offspring of females with chronic experimental liver damage of various etiologies. Morphology. 2006. - Volume 129, No. 1. - S. 57-59.
6. Zaitsev I.A. Viral hepatitis in women of reproductive age // Clinical Immunology. Allergology Infect ology. - 2017. - No. 5 (102). - C. 36-42. ISSN 24112852 www.kiai.com.ua
7. Zufarov K. A. & Tukhtaev K. R. (1987). Organs of the immune system. Moscow: FAN Publishing House.
8. Zufarov K.A., Tukhtaev K.R., & Khasanov B.B. (2003). Quantitative and ultrastructural characteristics of immunocompetent cells of the mammary gland in the dynamics of pregnancy and lactation. Morphology, 124(4), 74-79.
9. Kashchenko S. A. et al. The structure of the spleen of white rats of suckling age / Tauride Medical and Biological Bulletin 2013, volume 16, No. 1, part 1 (61). - P.104-106.
10. Otajonova A. N., Azizova F. Kh., & Tukhtaev, K. R. (2011). Effect of taktivin on the structural state of Peyer's patches in chronic toxic hepatitis. PhD student 45(2), 39-43.
11. Khasanov B. B. (2009). Structural and functional features of the mammary gland in the dynamics of pregnancy and lactation against the background of
toxic hepatitis. Medical business, Kiev "Health", (7-8), 94.
12. Azizova F. X. and others. (1997). at al. Structural and functional properties of mesenteric lymph nodes under antigenic influence in early postnatal ontogeny. Uzbekistan Medical Journal, (10-11), 14-16.
13. Baban B., Malik A., Bhatia J. and others. Presence and Profile of Innate Lymphoid Cells in Human Breast Milk. JAMA. 2018; 172(6):594-596. doi:10.1001/jamapediatrics.2018.0148
14. Burthanovich, K. B. (2021). Structural and functional features of immunocompetent breast cells glands during pregnancy and lactation in chronic hepatitis. Psychology and Education Journal, 58(2), 80388045.
15. Cesta M. F. (2006b). Normal structure, function, and histology of the spleen. Toxicol Pathol 34, 455-65.
16. Elmore, S. A. (2012). Enhanced histopathol-ogy of the immune system: A review and update. Tox-icol Pathol 40, 148-56.
17. George A. Parker and others. Histologic features of postnatal development of immune system organs in the sprague-dawley rat // Toxicologic Pathology. - 2015. - 43: 794-815, DOI: 10.1177/0192623315578720
18. Georgeuntzou A and N.G. papadopoulos. (2017) Postnatal Innate Immune Development: From Birth to Adulthood. Front. Immunol. August 11, 2017 | https://doi.org/10.3389/finunu.2017.00957
19. Karimov Kh.I. Effect of maternal toxic hepatitis on the functional characteristics of the lactation process. Likars'ka Sprava, Kiev, (5-6), 68.
20. 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.
21. Kh, A. F., Kh, B. D., & Kh, A. (2001). Age-related structural and functional features of the small
intestine of rats born from female rats with chronic toxic hepatitis. Medical business (1), 103-105.
22. 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.
23. Khasanov B. (2021). Maternal toxic hepatitis, structural and functional formation of the lean intestine of the offspring in the dynamics of early postnatal ontogenesis. The scientific heritage (78-2), 33-37.
24. Shilova I.V., Ostankova Yu.V., Goryacheva L.G., Semenov A.V. HBV infection in children with perinatal infection. Clinical report of familial hepatitis B. Journal of Infectology. 2021; 13(3):120-124. (In Russ.) https://doi.org/10.22625/2072-6732-2021-13-3-120-124
25. Sultanova D. (2021). The influence of mother's extra genital pathology on the formation spleens of offspring in the early period postnatal ontogenesis. The scientific heritage, (81-2), 47-49.
26. 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 the suckling period. Morfologiia (Saint Petersburg, Russia), 124(6), 70-72.
27. Volkov V.P. A new algorithm for the morpho-metric evaluation of the functional immunomorphol-ogy of the spleen. Universes: Medicine and pharmacology: electronic scientific journal 2015; 5-6 (18) [Electronic Resource]. URL: http://7universum. com/ru/med/archive/item/2341. (Date of access April 10, 2017).
28. Zufarov K.A. and other. (2003). Quantitative and ultrastructural characteristics of immunocompetent cells in the mammary gland during pregnancy and lactation. Morphology (Saint Petersburg, Russia), 124(4), 74-79.
