ОЦЕНКА МОРФОФУНКЦИОНАЛЬНЫХ ИЗМЕНЕНИЙ ПЕЧЕНИ ПРИ ДИАБЕТИЧЕСКОЙ ГЕПАТОПАТИИ

doi:10.28942/atuj.v6i3y2026.165

Абстракт

В статье представлены данные об изменениях маркеров нарушения метаболизма в печени (аланинаминотрансфераза — АЛТ, аспартатаминотрансфераза — АСТ, гамма-глутамилтрансфераза — ГГТ, лактатдегидрогеназа — ЛДГ) в крови и лимфе, о скорости лимфотока (СЛТ), а также о гистоморфологических особенностях печени при аллоксан-индуцированном сахарном диабете у кроликов. Экспериментальные исследования были проведены на 14 кроликах обоего пола. Животные были разделены на 2 группы: контрольную (6 кроликов) и экспериментальную (8 кроликов). Диабетические симптомы были выявлены на 5-й день после инъекции аллоксана. Для анализа брали кровь из полости сердца, а лимфу — из грудного лимфатического протока. Операции проводились под анестезией. У кроликов обеих групп исследовали уровень глюкозы, маркеры нарушения метаболизма печени, скорость лимфотока, а также гистоморфологические особенности печени. У животных экспериментальной группы показатели определялись на 5-й, 15-й и 30-й дни после введения аллоксана. Установлено, что повышение маркеров нарушения метаболизма печени как в крови, так и в лимфе, а также развивающиеся гистоморфологические изменения в печени сопровождались снижением скорости лимфотока

Сопроводительное письмо

Diabetes mellitus (DM) is a rapidly spreading disease worldwide, characterized by long-term complications, and remains one of the serious medical and social problems. According to the World Health Organization, in 2021, the number of people with diabetes worldwide exceeded 500 million, and it is projected that this indicator will reach 700 million by 2030 [ 1, 2, 3]. Diabetes is also spreading rapidly in Azerbaijan. In 2024, the total number of registered diabetic patients in the unified electronic registry of medical institutions under the Administration of the Regional Medical Divisions was reported to be 322,353 [4, 5 ].

One of the organs most affected by DM is the liver. The liver plays a key role in regulating glucose homeostasis. During persistent hyperglycemia, end-products of non-enzymatic glycosylation accumulate in the body, including hepatocytes. This leads to damage of both intracellular and extracellular structures, a decrease in NADPH levels, an acceleration of oxidative stress, and increased conversion of glucose to sorbitol [6, 7, 8, 9, 10, 11]. As a result of the disruption of the integrity of hepatocyte membranes, enzymes such as ALT, AST, GGT, and LDH are released into the blood and lymph. This is considered an indicator of liver damage. Although significant progress has been made in studying the histomorphological changes in the liver and the alterations in the levels of enzymes considered indicators of liver damage in the blood in diabetes mellitus, however, there is limited information about such changes in the lymph under conditions of persistent hyperglycemia [12, 13, 14, 15]. It should be noted that a significant portion of the lymph in the thoracic duct originates from the liver, and lymphatic fluid can provide more detailed information about metabolic processes occurring in tissues [6, 17, 18]^..Although the role of the lymphatic system in the pathogenesis of liver failure is of great importance, its role during diabetes mellitus has not been sufficiently investigated [19, 20, 21,22].

The aim of this research was to comparatively study the levels of markers impaired metabolism in the liver (ALT, AST, GGT, LDH) in the blood and lymph during experimentally induced diabetes mellitus in rabbits using alloxan injection, to determine the lymph flow rate, and to investigate histomorphological changes in the liver.

MATERIALS AND METHODS The study was carried out on 14 rabbits of the “Chinchilla” breed, weighing between 1.8 and 2.3 kg. The rabbits used in the experiments were divided into two groups: the control group included 6 animals, and the experimental group included 8. In the experimental group, diabetes mellitus was induced in rabbits by intraperitoneal administration of 5% alloxan monohydrate solution at a dose of 100 mg/kg. Diabetic symptoms were observed 5 days after the injection. Blood samples were obtained from the cardiac cavity and lymph samples from the thoracic duct. All procedures were carried out under anesthesia. For this purpose, 8 mg/kg of Calypsol and 0.15 ml/kg of 1% Dimedrol solution were injected intravenously. In the control group, blood and lymph samples were collected from six rabbits, and the levels of liver function–related enzymes (ALT, AST, γ-GT, LDH) as well as LFR were determined. In addition, the histomorphological structures of the liver were studied. The same parameters were assessed in the experimental group of 8 rabbits on 5th, 15th, and 30th days after alloxan administration. For histomorphological evaluation, liver tissues were fixed using 10% neutral buffered formalin or Bouin’s solution. Subsequently, the specimens were dehydrated through a graded ethanol series, embedded in paraffin, sectioned, and stained for histological examination. The numerical data obtained during the course of the study were statistically analyzed in accordance with current recommendations. Statistical analyses were carried out using the method of variation analysis and with the help of the ‘IBM Statistics SPSS-26’ software. For the comparison of variation series, the non-parametric Mann-Whitney U test was also used.

RESULTS AND DISCUSSION During the study, the levels of enzymes characterized the diabetic hepatopathy (ALT, AST, γ-GT, LDH), LFR, and the histomorphological structures of the liver were investigated in the blood and lymph of rabbits with alloxan-induced diabetes mellitus over a period of 30 days and compared with control values. The obtained results are presented in the tables below (Table 1).

As seen from Table 1, after alloxan administration, the levels of blood glucose and the liver function-assessing enzymes (ALT, AST, GGT, LDH) gradually increased throughout the experiment (p<0,001). The dynamics of changes in the parameters determined in the lymph are in the same direction as in the blood (Table 2). As seen from Table 2, after alloxan administration, the levels of glucose and liver function-assessing enzymes (ALT, AST, GGT, LDH) in the lymph gradually increased (p<0.001), and the lymph flow rate decreased throughout the experiment (Table 2).

As seen from the presented tables, in animals with induced diabetes mellitus, an increase in the activity of ALT, AST, GGT, and LDH in blood and lymph was observed throughout the experiment compared to the control values. This tendency was more pronounced in the lymph than in the blood. In rabbits with alloxan-induced diabetes mellitus, the increased markers of metabolic disorders in the liver (ALT, AST, GGT, LDH) in blood and lymph are explained by the acceleration of hepatocyte cytolysis under persistent hyperglycemia [23, 24, 25]. During the experiment, histomorphological changes developing in the liver were studied on the 5th, 15th, and 30th days after alloxan administration. On the 5th day of alloxan-induced diabetes mellitus, dystrophic changes were observed in hepatocytes around the periportal vessels (Figure 1). Based on the information presented above, we conclude that during diabetes mellitus, enzymes released from damaged hepatocytes enter the bloodstream and lymphatic system. The increase of these enzymes in both blood and lymph, along with the histomorphological changes occurring in the liver, is considered the development of diabetic hepatopathy, one of the long-term complications of diabetes mellitus [26, 27, 28, 29 ]. The decrease in lymph flow rate slows the removal of accumulated toxic products from tissues, further deepening the histomorphological changes that are developing in the liver [30, 31,32,33].

CONCLUSİON. In alloxan-induced diabetes mellitus, the increased markers of metabolic disorders in the liver (LDH, AST, ALT, GGT) in both blood and lymph, along with the progressive deepening of histomorphological changes in the liver, were accompanied by a decrease in lymph flow rate.

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