THE EFFECT OF BIOFLAVONOIDS ON THE FUNCTIONS OF VISCERAL LYMPH NODES IN DIFFERENT AGE PERIODS

ABSTRACT

The purpose of the work is a studying influence of Siberian herbs on a morphofunctional status of visceral lymph nodes of different regional belonging at young and old animals. Phytocomposition bioflavonoids are the main active ingredient. There are morphological and thermogravimetric methods in work. It is shown what features of the lymphatic region define distinctions of a morphofunctional status of mesenteric and tracheobronchial lymph nodes. Parameters of hydration are objective criterion of a morphofunctional condition of regional lymph nodes depending on localization and age. Aging causes involution of lymphoid tissue and dehydration of visceral lymph nodes with depression of function. Phytotherapy leads to strengthening of drainage and immune functions of visceral lymph nodes. Phytotherapy renders the structural modifying effect and increases hydration of the lymph nodes which especially underwent senile changes. The received results confirm the concept of the lymphatic region that should be considered at a research of lymph nodes during the different periods of life.

Keywords: lymph nodes, hydration, bioflavonoids, phytotherapy.

INTRODUCTION

Lymph nodes perform drainage and immune functions and have a number of morphofunctional features that obey the postulates of the concept of the lymphatic region [1, p. 12]. The function of the lymph nodes is inextricably linked to tissue metabolism and water balance in the region of the lymph catchment and changes with age [2, p. 139, 174]. The importance of water homeostasis, as well as the structure of the lymph nodes, is given great importance, since the lymph nodes are considered an important element of the body's liquid homeostasis. With aging, many morphofunctional parameters of lymph nodes are destabilized, and with them their function [1, p. 18, 24; 2, p. 249]. Therefore, it is natural to search for ways and means to optimize the work of the lymphatic system (including lymph nodes). One of such methods can be bioflavonoids of medicinal plants of Siberia, which have lymphotropic properties [1, p. 29; 2, p. 145, 179]. At the same time, the assessment of the state of regional lymph nodes in their relationship with drained organs during aging and after phyto-correction remains one of the main directions in morphophysiological studies.

The aim of the study is to evaluate the possibility of phytocomposition in optimizing the morphofunctional status of visceral lymph nodes, taking into account localization and age.

MATERIAL METHODS

The experiment was carried out on 80 white male Wistar rats with conditional allocation of age groups-"young" (3-5 months) and "old" (1.5 – 2 years). The animals received a standard diet (extruded feed) with free access to water and were controlled. In addition, for one month, the animals of different ages were given phytocollection pellets, including crushed medicinal plants. The average daily dose of phytocollection was 0.1–0.2 g/kg. Each group had at least 20 animals. The phytocomposition included medicinal plants – rhizomes and leaves of the tea copse (Hedysarum theinum Krasnob.), badan (Bergenia crassifolia (L.) Fritsch.), Rhodiola rosea (Rhodiola rosea L.), blueberry leaves (Vaccinium myrtillus L.), lingonberry (Vaccinium vitisidaea L.), black currant (Ribes nigrum L.), rosehip fruits (Rosa majalis Herrm.), the main active ingredient is bioflavonoids [3, p. 3-8; 7, p. 19].

Visceral lymph nodes (mesenteric and tracheobronchial) belonging to different lymphatic regions were selected as the object of the study.

Histological examination of lymph nodes. The collected lymph nodes were fixed in 10% neutral formalin. This was followed by the classical scheme of wiring and pouring the material into paraffin, followed by the preparation of histological sections. Histosections were stained with hematoxylin-eosin, a trichromic dye according to C. Masson, and subjected to morphometric analysis.

Thermogravimetric method [4, p. 32] for the assessment of hydration with the determination of different water fractions (total, free and bound) and the coefficients of hydration and density in the lymph nodes. The principle of the method is to dry the lymph node at a constant temperature of 105 C⁰ with the registration of changes in its mass on the analytical balance. Knowing the volume of the lymph node and the area of its structures, we calculated the volume of fluid that falls on a particular structure of the lymph node in accordance with the Cavalieri–Aker–Glagolev principle.

Statistics. Statistical processing of the study results was carried out using Excel and StatPlus Pro, AnalystSoft Inc. The arithmetic mean was calculated and its standard (root-mean-square) error was determined. The Student's t-test was used to assess the level of statistical significance of differences (p < 0.05) under the condition of a normal distribution or close to it.

RESULTS AND DISCUSSION

Structurally and functionally, the lymph nodes are tied to the territory that is drained, which is in accordance with the provisions of the concept of the lymphatic region [1, p. 12; 8, p. 1]. But the peculiarity of each lymphatic region, especially the conditions of its contact with the external environment, determines the morphofunctional status of the regional lymph node and its response to herbal medicine in different periods of life.

Mesenteric lymph node. The structural support of mesenteric lymph nodes ensures the performance of drainage and immune functions equally at a young age, when the lymphoid tissue has maximum development [1, p. 18; 2, p. 139; 5, p.22]. The intake of plant bioflavonoids affects the parameters of lymph node hydration and thereby alters its drainage activity. However, this effect manifests itself within the physiological values, without giving a statistically significant difference for most indicators (Table 1).

Table 1.

Mesenteric lymph node parameters before and after taking plant bioflavonoids in young animals (n=40)

Thus, a change in the hydration coefficient (by 1.5 times) indicates a redistribution of free and bound water fractions in the lymph node when taking plant bioflaonoids. The volume of the free fraction of water decreases by 1.2 times, the volume of the bound fraction of water increases by 1.3 times when taking the phytocomposition (Table 1). At the same time, the volume of intercellular fluid and the volume of lymph in the sufficiently wide lymphatic sinuses in the lymph node practically does not change. The hydration conditions created by taking the herbal medicine support the existing lymph transport and enhance the function of lymphoid cells in the intranodular zones of the lymph node of young animals.

Aging leads to age-related involution of the mesenteric lymph node lymphoid tissue with a decrease in immune and drainage functions. In the structure of the mesenteric lymph node, the predominance of medullary matter was noted mainly due to the B-dependent zone (brain cords) and evenly expanded lymphatic sinuses, while minimizing the area of lymphoid structures [1, p. 24; 2, p.139]. This reflects a weakened humoral immune response in the senile mesenteric lymph node. The value of the hydration coefficient indicates a pronounced dehydration of the mesenteric lymph node when compared in young and old animals (Tables 1, 2).

Table 2.

Mesenteric lymph node parameters before and after taking plant bioflavonoids by old animals (n=40)

Plant bioflavonoids change the size of the lymph node compartments, which is regarded as a positive response in old animals. The compartments that were reduced without correction increase, and, conversely, the structures that were increased decrease. This is regarded as a structural-modifying effect of herbal medicine [1, p. 29; 2, p.145]. As a result of taking plant bioflavonoids, the water mass of the mesenteric lymph node increases by 1.2 times with an increase of 1.5 times the volume of lymph and, consequently, the free fraction of water in the lymphatic sinuses of the lymph node (Table 2). The volume of extracellular fluid remains constant. The value of the hydration coefficient reflects changes in the water (liquid) balance in the mesenteric lymph node after taking plant bioflavonoids (Table 2). It is obvious that taking phytocomposition has a lymph-stimulating effect, optimizing the drainage function of the lymph node.

Tracheobronchial lymph node. In young animals, the lymph node is characterized by a certain structure, where it has a predominance of cortical matter in the presence of narrow lymphatic sinuses [1, p. 18; 2, p. 174; 5, p. 22]. The most developed thymus is an independent B-zone, including lymphoid nodules and brain cords, which indicates an immune response of the humoral type. The structure is reflected in the indicators of lymph node hydration. The density index has a rather high value due to the predominance of the cellular component in the lymph node (Table 3). Rather low indicators of hydration of the tracheobronchial lymph node have a physiological character, reflecting the peculiarity of the lymphatic region. The tracheobronchial lymph node of young animals is characterized by low indicators of fluid volume in the sinus system and water fractions, as well as the proportion of bound water in comparison with similar indicators of the mesenteric lymph node (Table 3).

Table 3.

Tracheobronchial lymph node parameters before and after taking plant bioflavonoids in young animals (n=40)

Taking plant bioflavonoids leads to an increase in water mass (1.5 times), lymph volume in the sinuses (1.3 times), intercellular fluid (1.9 times) and total water volume (1.3 times) due to the free water fraction (1.6 times) (Table 3). Such a change in hydration after phytotherapy enhances the drainage and immune functions of the tracheobronchial lymph node, determining the lymph-stimulating effect of plant bioflavonoids for this lymphatic region [2, p. 179].

With aging, the tracheobronchial lymph nodes undergo involution of the lymphoid tissue, leading to a decrease in the dimension of all structural and functional zones [1, p. 24; 2, p.174; 5, p.; 6, p. 19]. At the same time, there was an expansion of the area of the interstitial part of the cortex and brain cords (B-zone), paracortex (T-zone) against the background of narrow lymphatic sinuses, which determine a weakened immune response of a mixed type with limited drainage function during aging. Dehydration is a characteristic feature of an aging tracheobronchial lymph node (Table 4). There is a decrease in the mass of the liquid (by 3.2 times), the volume of intercellular fluid (by 3.3 times), the free fraction of water (by 1.7 times), but the volume of the bound fraction of water increases by 1.2 times (Tables 3, 4).

Table 4.

Tracheobronchial lymph node parameters before and after taking plant bioflavonoids by old animals (n=40)

It was shown that the use of the herbal medicine leads to an increase in the area of lymphoid nodules and the sinus system, which had minimal areas in the lymph node in old animals. Simultaneously improves the water saturation of the lymph node. The intake of plant bioflavonoids increases the mass of fluid (1.7 times), the volume of intercellular fluid (2.8 times) and the free fraction of water (1.4 times), and reduces by 1.6 times the volume of the bound fraction of water in the tracheobronchial lymph node subjected to aging (Table 4). Such changes indicate the lymph-stimulating (drainage and immune) effect of phytocomposition.

CONCLUSION

The peculiarities of the contact of the lymphatic region with the external environment determine the differences in the morphofunctional status of the mesenteric and tracheobronchial lymph nodes. Hydration parameters are an objective criterion of the morphofunctional state of regional lymph nodes, depending on the location and age. Aging causes the involution of lymphoid tissue and dehydration of visceral lymph nodes with a decrease in their function. Phytotherapy leads to increased drainage and immune functions due to the structural-modifying effect and increased hydration of lymph nodes, especially those that have undergone senile changes. The obtained results confirm the main provisions of the concept of the lymphatic region, which should be taken into account in the prognostic assessment when examining the lymph nodes in different periods of life.

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