动物造模 z-bio 2022-01-04 256

　　Objective To study the molecular mechanism of stress affecting appetite from the ghrelin pathway.

　　Method 32 male Wistar rats were randomly divided into a control group and a stress group, with 16 in each group and 2 in each cage. The tail-clamping stress method was used to stimulate the rats twice a day for 21 consecutive days to monitor the food intake and body weight of the rats in each group; 8 rats in each group were selected on the 7th and 21st days of the experiment, and the serum was detected by ELISA The mass concentration of acylated ghrelin; Western blotting was used to detect the expression of ghrelin protein in the rat stomach and hypothalamus, and the expression level of adenylate-activated protein kinase α2 (AMPKα2) in the hypothalamus; real-time fluorescent quantitative PCR detection Ghrelin, growth hormone secretagogue receptor (GHSR), leptin (leptin) in the stomach, and ghrelin, AMPKo2, neuropeptide Y (neuropeptide Y, NPY), and agouti gene-related protein in the hypothalamus ( agouti-related protein (AgRP) mRNA level.

　　Results The total food intake of rats in each cage in the stress group was more than that of the control group on the 7th day of the experiment (P<0.05), and="" there="" was="" no="" significant="" difference="" in="" the="" total="" food="" intake="" of="" two="" groups="" rats="" on="" 21st="" day="" p="">0.05); On day 21, there was no significant difference in body weight between the two groups of rats (P>0.05). Stress can significantly up-regulate the serum acylated ghrelin level (P<0.01), and the expression of ghrelin protein (P<0.05) in the stomach and hypothalamus and AMPKg2 protein (P<0.01) in the hypothalamus were also significantly up-regulated. Stress can also significantly up-regulate the levels of ghrelin and GHSR mRNA in the stomach (P<0.01), and down-regulate the levels of leptin mRNA in the stomach (P<0.01); at the same time, it can up-regulate the levels of ghrelin, AMPKα2, NPY and AgRP mRNA in the hypothalamus (P≤0.01) .

　　Conclusion Tail-clamping stress can cause an up-regulation of ghrelin levels in the center and periphery of rats, which may have an effect on the temporary increase in appetite in rats.