УДК 542.934:572.7:577.17.049:611:428:612.67

Possibilities of phytocorrection of morphofunctional state of lymph nodes at different ages

Веременко Анастасия Сергеевна – студент Института медицины и психологии имени Зельмана Новосибирского государственного университета.

Левченко Ирина Дмитриевна – студент Института медицины и психологии имени Зельмана Новосибирского государственного университета.

Горчаков Владимир Николаевич – доктор медицинских наук, профессор Научно-исследовательского института Клинической и экспериментальной лимфологии – филиала Института цитологии и генетики Сибирского отделения Российской академии наук.

Вергунова Екатерина Евгеньевна – студент Института медицины и психологии имени Зельмана Новосибирского государственного университета.

Тумас Артем Сергеевич – студент Института медицины и психологии имени Зельмана Новосибирского государственного университета.

Бекенева Ксения Алексеевна – студент Института медицины и психологии имени Зельмана Новосибирского государственного университета.

Новикова Мария Федоровна – студент Института медицины и психологии имени Зельмана Новосибирского государственного университета.

Шаймарданова Диана Рамильевна – студент Института медицины и психологии имени Зельмана Новосибирского государственного университета.

Abstract: This study focuses on the need to stimulate lymphatic drainage and optimise morphofunctional parameters of lymph nodes at different stages of ontogenesis. This aspect acts as a fundamental direction in research work combining the principles of lymphology and gerontology. The main focus of the work is to improve the efficiency of lymph nodes through the application of phytotherapy methods. In this context, bioflavonoids of plant origin, which have significant therapeutic potential, play a special role. The study was conducted to evaluate the effect of an innovative phytopreparation on the morphological and functional state of lymph nodes in rats of different ages, including young and elderly individuals. The results showed that the use of phytotherapy has a positive effect on the structure and functional characteristics of lymph nodes, thereby improving their performance and efficiency throughout different life stages. These findings represent a significant contribution to the development and improvement of wellness and rehabilitation programmes aimed at maintaining health and slowing down ageing processes, and can be used as a basis for future medical practices and therapeutic approaches. Thus, this study opens new perspectives on the use of phytopreparations to improve the health of the lymphatic system and offers innovative approaches for the treatment and prevention of diseases associated with lymph node dysfunction. It also emphasises the importance of integrating knowledge from different fields of medicine to achieve a comprehensive understanding and improve health across the lifespan.

Аннотация: В рамках данного исследования уделяется внимание необходимости стимулирования лимфатического дренажа и оптимизации морфофункциональных параметров лимфоузлов на разных этапах онтогенеза. Этот аспект выступает в качестве фундаментального направления в исследовательской работе, сочетающей принципы лимфологии с геронтологией. Основное внимание в работе уделено повышению эффективности работы лимфоузлов через применение методов фитотерапии. В этом контексте особую роль играют биофлавоноиды растительного происхождения, которые обладают значительными терапевтическими возможностями. Исследование проводилось с целью оценить влияние инновационного фитопрепарата на морфологическое и функциональное состояние лимфатических узлов у крыс различного возраста, включая молодых и пожилых особей. Результаты показали, что использование фитотерапии оказывает положительное воздействие на структуру и функциональные характеристики лимфоузлов, тем самым улучшая их работоспособность и эффективность на протяжении разных жизненных этапов. Эти открытия представляют собой значительный вклад в разработку и усовершенствование оздоровительных и реабилитационных программ, направленных на поддержание здоровья и замедление процессов старения, и могут быть использованы в качестве основы для будущих медицинских практик и терапевтических подходов. Таким образом, данное исследование открывает новые перспективы в области применения фитопрепаратов для улучшения состояния лимфатической системы и предлагает инновационные подходы в области лечения и профилактики заболеваний, связанных с нарушениями в работе лимфатических узлов. Оно также подчеркивает важность интеграции знаний из различных областей медицины для достижения комплексного понимания и улучшения здоровья на протяжении всей жизни.

Keywords: bioflavonoids, phytostimulation, lymph nodes, lymphatic region.

Ключевые слова: биофлавоноиды, фитостимуляция, лимфатические узлы, лимфатический регион.

Introduction

During ontogenesis, the human body is exposed to multiple exogenous and endogenous influences that can destabilise the structure and function of organs and systems at any age. There is an exhaustion of compensatory mechanisms, which, in turn, increases the risk of initiation of pathological processes. Over the years, this risk only increases due to the continuing action of harmful environmental factors and a decrease in the regenerative and adaptive abilities of the body. A special role is assigned to the lymphatic system and, in particular, to lymph nodes in ensuring the body's defence against the action of exo- and endogenous factors at different ages [1]. Structural changes of lymph nodes induced by pathogenetic factors in adolescence or age-related changes in old age may be accompanied by impairment of their function, which in the long term may lead to reduced quality of life. Understanding the mechanisms of negative factors impact on the lymphatic system is the subject of scientific research and is of paramount importance for the development of a strategy for prevention and correction of emerging disorders.

The need to correct the structure and function of the lymphatic system requires the search for methods and means aimed at improving the efficiency of the lymph nodes in the lymphatic region. The method of choice is phytotherapy, which allows to achieve stimulation of lymphatic drainage and increase the immune function of peripheral lymphoid organs [2,3]. It is debated to what extent it is reasonable to apply phytotherapeutic methods in the conditions of decreasing activity of lymphoid tissue with age. Nevertheless, from the position of preventive gerontology and lymphology, phytotherapy represents an important direction in research related to the protection and counteraction to aging of the lymphatic (lymphoid) system [4,5].

The aim of the study was to investigate the effect of the original phototherapy on the morphofunctional characteristics of lymph nodes at different stages of ontogenesis.

Literature review

Phytocorrection, as an innovative approach in modern medicine, is a method of influence on the body using plant preparations. This method becomes especially relevant when considering the morphofunctional state of lymph nodes in different age periods. The studies of Yu.M. Levin [2] emphasise the importance of stimulation of interstitial humoral transport and lymphatic drainage of tissues. These processes play a key role in ensuring the normal functioning of the lymphatic system. Based on these findings, a detoxification methodology known as atraumatic endoecological rehabilitation according to Levin (AERL) has been developed [2, 6]. This method involves active intervention to improve the transport processes occurring in the interstitial space and lymphatic channel.

A key role in maintaining health and preventing various diseases is attributed to plant bioflavonoids as pharmacologically active compounds [7]. Their antioxidant, anti-inflammatory and immunomodulatory properties make them valuable ingredients in plant bioactive supplements of functional foods. Plant bioflavonoids are attracting increasing attention from scientists due to their potential health and therapeutic properties, especially in the context of enhancing the function of the lymphatic system. In particular, natural bioflavonoids such as quercetin and other plant-derived polyphenolic compounds have shown promising results in experiment [8]. These compounds may have antioxidant, anti-inflammatory effects and reduce oedema by improving lymphatic circulation, [9]. Bioactive supplements ("Lymphosan", "Lymphovit", "Harmonia Vita") containing Siberian plants deserve attention [10]. Their use in sanatoriums of the Altai region showed high efficiency, especially in the older age group, which confirms their potential in the correction of morphofunctional state of lymph nodes in the implementation of the principles of lymphostimulation, lymphoprotection and lymphocorrection. The literature describes the properties of various phytomedicines and their ability to influence the lymphatic system, and in particular the lymph nodes [6, 11]. The use of plants such as common cuff, medicinal bloodroot and others helps to improve the function of the lymphatic system [6,11]. These plants have unique properties to stimulate lymphatic drainage and improve lymph node health.

Despite extensive research in this area, many aspects of the effects of phytomedicines on the lymphatic system remain unexplored. In particular, the mechanisms of interaction between flavonoids and different cell populations of lymph nodes, as well as their role in modulating the immune response, require further investigation. Our data confirm the ability of flavonoids to exert corrective and stimulating effects on lymph nodes at different stages of ontogenesis in accordance with the concept of lymphatic region by Yu.I. Borodin [16].

Materials and Methods

The study focused on investigating three topographic groups of lymph nodes in rats. These are inguinal, mesenteric and tracheobronchial nodes. The choice of these groups was made based on the principles of ecological lymphology [4,5].

To obtain the most accurate and objective data, white male rats of Wistar breed were chosen as the objects of the study. The age of these animals was 5 months (young) and 1.5 years (old). The age of rats is correlated with the age of humans. Thus, 5 months of rat life equates to 16-20 years of human life, while 1.5 years of rat life corresponds to 75 years of human life based on a coefficient of 1.7 [12]. During the experiment, the animals were divided into control and experimental groups. The control group included young and old animals without phytoremediation and the experimental group included young and old animals receiving the phytoremediation. The phytoremediation was added to food with standard diet. There were at least 10 animals in each group.

The phytopreparation formulated as biologically active additive IQdetoxSORB (NPF "SIB-CRUK", Koltsovo, Novosibirsk) was used. IQdetoxSORB is a powder of mechanochemical grinding of plants (flax seeds, tea kettle (Hedysarum theinum Krasnob), thick-leaved badanum (Bergenia crassifolia), May rosehip (Rosa majalis Herrm. ), blackcurrant (Ribes nigrum L.), Rhodiola rosea, blueberry myrtillus (Vaccinium myrtillus), common lingonberry (Vaccinium vitis-idaea), thyme creeping (Thymus serpyllum L.) [13]. The daily dose of the combined phytopreparation was 200 mg/kg. Its most important bioactive substances are flavonoids, arbutin, dietary fibre and trace elements, allowing to achieve antioxidant, anti-inflammatory, sorption and protective effects.

Light microscopy methods were used for detailed analysis of lymph node morphology. Histological analysis of lymph node structure was performed in accordance with the established standards and recommendations [14,15]. Leica DM 750 and Mikmed-2 microscopes equipped with ScanMicro specialised scanning attachment were used in the study.

Own results

Young animals. Phytotherapy affects the structure of lymph nodes in young animals. Plant bioflavonoids, acting on cellular immunity, change the structural organisation of lymph nodes. After this therapy, cortical matter in lymph nodes occupies 64.72%-69.48% and brain matter 30.46%-32.01%. The index of cortical-brain ratio exceeds two, being 2.12±0.04 for inguinal and tracheobronchial lymph nodes and 2.28±0.06 for mesenteric lymph nodes.

After phytotherapy in young animals there are changes in the dimensionality of intranodal zones in the lymph node structure. In the tracheobronchial lymph node, the internodal cortical area increases to 14.24±0.70%, which is 1.9-2.1 times larger than in the inguinal and mesenteric lymph nodes. The paracortical area varies from 26.8% to 32.2%, with no statistical differences between lymph nodes. Thymus-dependent area is 38.81% in inguinal, 41.78% in tracheobronchial and 34.09% in mesenteric lymph nodes.

The area of lymphoid nodules remains high after phytotherapy, being 5.72%-8.26% in the absence of germinative centres and 9.89%-12.63% in the presence of germinative centres. The ratio of secondary to primary lymph nodes is 1.35±0.04 and 1.53±0.06 in inguinal and mesenteric lymph nodes, respectively, and 2.15±0.18 for tracheobronchial lymph nodes.

The area of medullary masses after phytotherapy was 25.33±1.47% for inguinal and 25.79±0.87% for tracheobronchial lymph nodes, and 18.15±0.66% for mesenteric lymph nodes. Thymus-independent area predominated over thymus-dependent area, indicating activation of the humoral link of cellular immunity. After phytotherapy, the sinus area increases, especially in the mesenteric lymph node, exceeding 1.8-2.2 times the values of other lymph nodes.

The effect of phytotherapy in young animals enhances drainage and immune functions of lymph nodes, minimally affecting their structure.

Old animals. Phytotherapy used in old animals has an effect on the structural organisation of lymph nodes of different regions. This is due to the fact that flavonoids contained in plants activate drainage and correct the structure of lymph nodes [8].

After phytotherapy in old animals, it was observed that the cortical area in inguinal and tracheobronchial lymph nodes is 72.68±2.12% and 69.23±2.71% respectively. These values are slightly less in mesenteric lymph node where they are 63.66±3.08%. Significant differences are observed between mesenteric and inguinal lymph nodes. When compared with the group of animals that were not corrected, after phytotherapy, the cortical area increases by 13.14% in inguinal lymph node, 8.12% in mesenteric lymph node, and decreases by 6.55% in tracheobronchial lymph node.

The mean brain matter area ranged from 24.82% to 36.26% in the lymph nodes of different regions, with the lowest in the tracheobronchial lymph node. After phytotherapy, the reduction in brain matter is 3-4% in inguinal and tracheobronchial lymph nodes and the highest reduction (11.47%) is observed in mesenteric lymph node.

After phytotherapy in aged animals, the inter-nodal cortical area remains smaller in all lymph nodes. Specifically, after phytotherapy, the area of the inter-nodal cortical part is 4.32±0.42% in inguinal lymph node, 6.99±0.31% in mesenteric lymph node, and 12.3±0.30% in tracheobronchial lymph node. Compared to animals without correction, the inter-nodal cortical area decreased 1.9-fold in inguinal lymph node, 1.2-fold in tracheobronchial lymph node, and increased 1.2-fold in mesenteric lymph node after phytotherapy.

After phytotherapy in older animals, the largest paracortical area is observed in the inguinal lymph node and is 43.1±2.18%. This is twice as much as in mesenteric (20.04±1.07%) and tracheobronchial (21.15±2.17%) lymph nodes.

Thymus-dependent area had the largest area in inguinal lymph node (47.42±3.12%), the smallest in mesenteric (27.03±3.17%) and medium in tracheobronchial (33.45±3.08%) lymph nodes. Administration of the phytocompound increased the area of lymphoid nodules with germinative centre in inguinal, mesenteric and tracheobronchial lymph nodes. After phytotherapy in aged animals the area of cerebral thrusts practically does not change in inguinal and tracheobronchial lymph nodes, but in mesenteric lymph node decreases by 1.3 times.

It was observed that after phytotherapy the common sinus system changes its lumen. In inguinal and tracheobronchial lymph nodes it approximates in size, being 10.84-12.59%. This is less than that of the mesenteric lymph node. This indicates that phytotherapy can modulate the structure of lymph nodes, making them more compact and functional.

After phytotherapy of aging lymph nodes, new structures appear, so-called - tertiary lymphoid nodules. The appearance of these lymphoid nodules is due to the need to compensate for the decrease in immune function and they are formed in certain situations. In this case, their formation was stimulated by phytotherapy.

Finally, after phytotherapy, differences in the structure of lymph nodes depending on their localisation were found in older animals. For example, the mesenteric lymph node had the lowest cortical-brain index, whereas the inguinal lymph node had a high thymus-dependent area. These differences may indicate the speciality of each lymph node and how phytotherapy may affect them.

Discussion

In the course of our study the fact of positive effect of phytotherapy on morphofunctional state of lymph nodes of different ages has been established. These results are confirmed in recent studies in accordance with the presented concept of lymphatic region [16]. Emphasis is placed on the importance of regional lymph nodes in the overall state of the lymphatic system. This is consistent with our observations on the effects of phytotherapy on regional lymph nodes and emphasises the importance of studying them in the context of prevention and treatment.

Ross and Kasum [17] conducted an in-depth study of plant flavonoids, looking at their bioavailability, metabolic effects and safety. In addition, the authors emphasised that the metabolic effects of flavonoids may contribute to their anti-inflammatory, antioxidant and cardioprotective effects. Plant flavonoids are effective on various body systems. The lymphatic system is not an exception. We have proved in experiment the ability of flavonoids to stimulate and maintain morphofunctional parameters of lymph nodes, especially those undergoing senile transformation.

One of the studies shows the ability of the flavonoid baicalein to induce proliferation and activation of T-regulatory cells (CD4(+)Foxp3(+) T-cells), which is a sign of a positive effect on the proliferative function of lymph nodes [18]. In turn, the application of the original phytopreparation leads to an increase in secondary lymphoid nodules and the appearance of tertiary lymphoid nodules as evidence of activation of differentiation and proliferation of lymphoid cells weakened by aging. It is evident that plant bioflavonoids can stimulate the proliferative activity of lymph nodes, especially at late stages of ontogenesis. A study by Khaliel et al. (2021), showed the ability of flavonoids to interact with sphingosine-1-phosphate receptors to enhance the migration of immune cells from lymph nodes into the bloodstream [21]. This effect may contribute to the optimisation of the body's systemic immune response and is evidence of the immunomodulatory activity of flavonoids.

Phytonutrients, increasing the performance of lymph nodes, slow down the aging process of lymphoid tissue [19, 20]. The obtained facts serve as a basis for the promising use of phytotherapy in health-preventive anti-aging programmes. Achievement of a positive effect is possible at realisation of principles of lymphosanation in order to provide effective work of lymph nodes at ageing. An original phytomineral complex combining stimulating and protective lymphotropic properties in relation to the lymphatic system is proposed. The phytocomposition has a positive effect on lymph nodes that have undergone senile changes [20]. And other scientists confirm the positive effects of phytotherapy on the lymphatic system by stimulating lymphatic drainage and improving the metabolism between cells and interstitial fluid [9, 22].

The effectiveness of phytocorrection for optimising the morphofunctional state of lymph nodes at different periods of life is becoming evident. Phytotherapy is a safe and effective way to influence the lymphatic system, especially in the elderly, and deserves further research and application in clinical practice.

Conclusions

Phytotherapy based on plant bioflavonoids changes the structural organisation of lymph nodes depending on age. Phytotherapy little changes the structure of lymph nodes with a tendency to increase the T-dependent zone during the period of maximum development of lymphoid tissue in young animals.

After phytotherapy, there is an optimisation of the structure and function of lymph nodes that have undergone age-related changes. In old animals phytotherapeutic effect is manifested in activation of lymphoid cells proliferation against the background of increase of the main structural and functional zones responsible for cellular and humoral immunity. One of the important points is the formation of extracortical substance - tertiary lymphoid nodules initiated by phytotherapy. 

Features of structural changes of lymph nodes depend on their topographo-anatomical localisation, which determines the specificity of the modifying effect on phytotherapy.

References

1. B-cell homeostasis and follicle confines are governed by fibroblastic reticular cells / V. Cremasco, M.C. Woodruff, L. Onder [et al.] // Nat. Immunol., 2014. – V. 15. – P.973–981.
2. Левин,Ю.М. Варианты Катрэла, Левинасан и новые препараты ЭРЛ / Ю.М. Левин // Материалы III Междун. конгресса «Эндоэкологическая медицина». Республика Кипр, 21-28 октября 2007 г. – М.: ОАО «Щербинская типография», 2007. – С.68–69.
3. Открытие свойства влиять на транспорт тканевой жидкости и лимфатический дренаж тканей у некоторых известных лекарственных препаратов / Ю.М.Левин, С.Т. Топорова, Л.П. Свиридкина, Ф.А. Баркинхоева [и др.] // Материалы III Междун. конгресса «Эндоэкологическая медицина». Республика Кипр, 21–28 октября 2007 г. – М.: ОАО «Щербинская типография», 2007. – С.69.
4. Бородин,Ю.И. Лимфатическая система и старение / Ю.И. Бородин // Фундаментальные исследования, 2011. – № – С.11–15.
5. Бородин,Ю.И. Эффекты лимфокоррекции и лимфопротекции при термическом ожоге кожи / Ю.И. Бородин, Н.П. Бгатова // Тезисы IV съезда лимфологов России. – М.: Проспект, 2011. – С.12-13.
6. Левин,Ю.М. Прорыв в эндоэкологическую медицину. Новый уровень врачебного мышления и эффективной терапии / Ю.М. Левин. – М.: ОАО «Щербинская типография», 2006. – 200 с.
7. West, B. , Deng, S. and Palu, '. (2020) Vitamin C, Grape Seed Extract and Citrus Bioflavonoids Protect the Skin against Photoaging: A Review. Journal of Biosciences and Medicines, 8, 116-134. 
8. Borodin IuI, Zykov AA. Farmakologicheskie sredstva, stimuliruiushchie drenazhnuiu funktsiiu limfaticheskoĭ sistemy [Pharmacologic agents that stimulate the drainage function of the lymphatic system]. Farmakol Toksikol. 1989 Mar-Apr;52(2):106-10.
9. Зыков,А.А. Фармакологические свойства биофлавоноидов и их влияние на адаптивно-компенсаторные процессы при ишемических повреждениях : Автореф. дис. … канд. мед. наук: 14.03.06 / Зыков Альберт Андреевич. – Казань, 1993. – 38 с.
10. Программа оздоровительных мероприятий по лимфосанации и детоксикации организма в ГУ НИИ клинической и экспериментальной лимфологии СО РАМН / Ю.И.Бородин, В.В. Асташов, В.Н. Горчаков, Т.А. Асташова [и др.]. – Новосибирск: ИПП «Манускрипт», 2004. – 70 с.
11. Горчаков,В.Н. Функциональная морфология почки и регионарного лимфоузла с характеристикой микроэлементного профиля при почечной недостаточности и фитокоррекции / В.Н. Горчаков, С.И. Анохин. – Saarbrücken: Palmarium Academic Publishing, 2012. – 175 с.
12. Гелашвили, О.А. Вариант периодизации биологически сходных стадий онтогенеза человека и крысы / О.А. Гелашвили // Саратовский научно-медиц. журнал, 2008. – Том 4. – № 22. – С.125–126.
13. Корсун, В.Ф. Фитотерапия. Традиции российского травничества / В.Ф. Корсун, Е.В. Корсун. – М.: Эксмо, 2010. – 880 с.
14. Белянин, В.Л. Диагностика реактивных гиперплазий лимфатических узлов / В.Л. Белянин, Д.Э. Цыплаков. – Санкт-Петербург-Казань: Типография издательства «Чувашия», 1999. – 328 с.
15. Cottier, H. Предложения по стандартизации описания гистологии лимфатического узла человека в связи с иммунологической функцией / H. Cottier, J. Turk, L. Sobin // Бюлл. ВОЗ. – 1973. – С.372–377.
16. Бородин Ю.И., Горчакова О.В., Суховершин А.В., Горчаков В.Н. и др. Концепция лимфатического региона в профилактической лимфологии. LAP LAMBERT Academic Publishing. 2018.
17. Ross J.A., Kasum C.M. Dietary flavonoids: bioavailability, metabolic effects, and safety. Annu. Rev. Nutr. 2002.
18. Bae MJ, Shin HS, See HJ, Jung SY, Kwon DA, Shon DH. Baicalein induces CD4(+)Foxp3(+) T cells and enhances intestinal barrier function in a mouse model of food allergy. Sci Rep. 2016 Aug 26;6:32225. doi: 10.1038/srep32225. PMID: 27561877; PMCID: PMC4999817.
19. Gorchakov V., Gorchakova O., Nurmakhanova B., Demchenko G. "Role of phytonutrients in delaying aging of somatic lymph node", Morphology, pathology, physiology, 13 (2), 2023
20. Gorchakov, Y. Kolmogorov, O. Gorchakova, K. Nikolaychuk, G. Demchenko and K. Bekeneva, "Principles of Lymphosanation in Ensuring the Efficiency Lymph Nodes at Aging," 2022 IEEE International Multi-Conference on Engineering, Computer and Information Sciences (SIBIRCON), Yekaterinburg, Russian Federation, 2022, pp. 290-294
21. Khaliel A.H., Abbas A., Hatem A., Abdulamir A. Sphingosine -1- phosphate receptor 1 polymorphism as a cause of Fingolimod unresponsiveness and Multiple Sclerosis initiation. Ann Trop Med & Public Health. 2021
22. Обмен биометаллов и электролитов при остром инфаркте миокарда и способы коррекции / А.В.Ефремов, А.Р. Антонов, Т.А. Литвинова, Я.Б. Новоселов [и др.] // Материалы Всерос. научно-практ. конф., посвящ. памяти проф. Ю.Г. Целлариуса «Клинико-морфологические аспекты общепатологических процессов при социально-значимых заболеваниях». – Новосибирск, 2004. – С.99.