Effect Of Zhiweifuwei Pills On HIF-1α And VEGF Expression In Rats With Precancerous Lesions Of Qi Deficiency And Blood Stasis

Objective:In this study,the rat model of gastric precancerous lesion（PLGC）was prepared by MMNNG solution compound method.After 4 weeks of drug intervention,the mRNA and protein expressions of hypoxia-inducing factor（HIF-1α）and angiogenesis factor（VEGF）in serum and gastric mucosa of rats in each group were observed.To explore the improvement mechanism of Zhiweifuwei Pills on gastric mucosa of rats with gastric precancerous lesions based on the treatment of qi and blood stasis,and to provide a new idea for clinical treatment of gastric precancerous lesions.Method:100 male SD rats were randomly divided into blank group（10）and modeling group（90）.The PLGC rat model was induced by N-methyl-N-nitro-n-nitrosoguanidine（MMNG）solution compound method for 26 weeks.At week 27,the modeling rats were randomly divided into model group,folic acid group,high,medium and low dose groups of Zhiweiwei Wan,and treated by intragastric administration for 4 weeks.After the last administration,the rats were fast for 24h and anesthetized by intraperitoneal injection of 2%pentobarbital sodium,abdominal aorta blood was taken from the rats,centrifuged at 3000r·min^-1 for 15 min,supernatant was taken and stored in the refrigerator at-80℃,and then the rats were placed on an anatomical plate,the whole stomach was dissected,and the stomach contents were rinsed with normal saline.Gastric tissues were spread out on filter paper to observe the damage of gastric antrum,gastric body and gastric Angle mucosa.Strip tissues（1cm×2mm）were taken from the side near the lesser curvature of the stomach to the gastric antrum,fixed in 4%paraformaldehyde and stored at room temperature.The remaining fresh tissues were fixed and storedin the-80℃refrigerator for later use.Hematoxylin-eosin（HE）staining was used to observe the histopathological changes of gastric mucosa in rats.The expressions of HIF-1αand VEGF protein and mRNA in gastric mucosa were detected by immunohistochemistry and Real-time PCR.Serum levels of angiogenesis related factors were detected by ELISA.The expressions of HIF-1αand VEGF were detected by immunofluorescence.Result:1.Pathological changes of gastric tissue under light microscope Zhiweifuwei Pills can significantly improve the pathological manifestations of gastric mucosal epithelial gland structure,arrangement disorder and dysplasia in PLGC rats.2.Expressions of HIF-1αand VEGF in gastric tissues of rats.RT-PCR results showed that compared with normal group,mRNA expressions of HIF-1αand VEGF in gastric mucosa of rats in model group were significantly increased（P<0.05）.Compared with model group,mRNA expressions of HIF-1αand VEGF were significantly decreased in Zhiweifuwei pills medium dose group（P<0.05）,and significantly decreased in high dose group（P<0.01）.Immunohistochemical results showed that compared with normal group,the expressions of HIF-1αand VEGF in gastric mucosa of rats in model group were significantly increased（P<0.05）.Compared with model group,the protein expressions of HIF-1αand VEGF in Zhiweifuwei Pills medium dose group were significantly decreased（P<0.05）,and those in high dose group were significantly decreased（P<0.01）.Immunofluorescence results showed that compared with normal group,the positive density values of HIF-1αand VEGF in gastric mucosa tissues of rats in model group were significantly increased（P<0.05）.Compared with model group,the positive density values of HIF-1αand VEGF were significantly decreased in Zhiweifuwei Pills medium-dose group（P<0.05）,and significantly decreased in high-dose group（P<0.01）.3.HIF-1αand VEGF expression in serum of rats ELISA results showed that compared with normal group,the expressions of HIF-1αand VEGF in serum of rats in model group were significantly increased（P<0.05）.Compared with model group,the expressions of HIF-1αand VEGF in serum of rats in Zhiweifuwei Pill medium dose group were significantly decreased（P<0.05）,and those in high dose group were significantly decreased（P<0.01）.Conclusion:Zhiweifuwei pills can significantly improve the pathological manifestations of gastric mucosal epithelial tissue abnormalities in rats with gastric precancerous lesions,and the mechanism may be related to down-regulating the overexpression of HIF-1αand VEGF and improving the hypoxia microenvironment.