QUANTITATIVE ANALYSIS OF REG1Α AND TGFB1 GENE EXPRESSION IN GASTRIC MUCOSA CELLS OF RATS UNDER THE EFFECTS OF IMMOBILIZED-WATER IMMERSION STRESS
DOI:
https://doi.org/10.31861/biosystems2024.02.227Keywords:
stress, stomach ulcer, RT-qPCR, Reg1a, Tgfb1 gene expressionAbstract
Gastric ulcer disease is the most common disease of the gastrointestinal tract, with a mortality rate of 5-10 % worldwide. The aim of the work was to quantitatively analyze the level of expression of genes Reg1a (encodes a regenerative protein) and Tgfb1 (encodes a multifunctional cytokine) in rat gastric mucosa cells under the influence of experimental immobilization water-immersion stress. Rats were removed from the experiment after 0.5, 1, 2, 3 hours of exposure to stress and after 12 and 24 hours after its withdrawal. Gene expression was studied on the isolated total RNA of rats using RT-PCR in real time (RT-qPCR), the results of which were calculated by the relative comparative method ("ΔΔCT Method"). The intensity of superoxide anion-radical production was determined by the accumulation of CTT-formazan. The protein content was determined by the Lowry method. An increase in the expression of both the Reg1a gene (at 1, 2, 3 h) and Tgfb1 (at 0.5, 1, 2, and 3 h) was detected against the background of an increase in the content of superoxide anion at 0.5, 1, 2, and 3 h under time of stressful exposure. At the same time, during the healing of ulcers, the expression of these genes approached the control values at 24 h after stress for Reg1a; and at 12 h for Tgfb1 (the level of superoxide anion radical formation returned to control at 12 h). The presence of a positive correlation between the expression patterns of these genes may indicate their pleiotropic effect during the regeneration of ulcerated gastric mucosa cells of rats.
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