Effect Of Zhuanggan Zhuyu Decoction On MST1,YAP And LATS1 Expression In Rats With Hepatic Fibrosis

Objective: In this study,the rat model of liver fibrosis was used as the research object to explore the effects of Zhuanggan Zhuyu decoction on the expression of MST1,YAP and LATS1 in rats with liver fibrosis,and further explore the mechanism of zhuanggan Zhuyu decoction against liver fibrosis,so as to provide objective basis for zhuanggan Zhuyu Decoction to prevent and treat liver fibrosis.Methods: Sixty SD rats were fed adaptively for one week,and 50 rats were randomly selected to construct liver fibrosis model,and the remaining 10 rats were blank group.Subcutaneously inject 40% CCl4 peanut oil solution,the first injection amount is 0.5ml/100 g,and the next injection amount is 0.3ml/100 g,three times a week.Take 5 model rats for He and Masson staining at the 6th and 8th weeks respectively to observe whether the model is successful.After the model was successful,the rats were randomly divided into model group,colchicine group,zhuanggan Zhuyu Decoction high-dose,medium-dose and low-dose groups(ZGZYJ high-dose,medium-dose and low-dose groups).The intragastric dose of ZGZYJ high,medium and low dose groups was 26.46 g/kg,13.23 g/kg and 6.615 g/kg,respectively.The intragastric dose of colchicine group was 0.126 mg/kg.The blank group and model group were given the same amount of normal saline,once a day,for 6 weeks.After the last administration of each group,the rats were fasted for 12 hours,weighed and recorde.After anesthesia,blood was taken from abdominal aorta,which was left standing at room temperature for 2h,and the upper serum was centrifuged to detect ALT,AST,ALB,ALP and other liver function indicators with automatic biochemical analyzer.Take rat liver,observe the morphology of liver tissue,and take two parts of right liver tissue,one part is fixed in 10% formaldehyde,and one part is frozen in the refrigerator at – 80 ℃.The fixed liver tissue was made into pathological sections,and HE and Masson staining were performed to observe the pathological morphology of the liver tissue under a microscope.The liver tissues frozen at-80 ℃ were detected by WB and RT-PCR,and the protein and m RNA expression levels of MST1,YAP and LATS1 in the liver tissues were observed.Results:1.general conditions of rats The hair color and glossiness of the rats in the drug treatment group were better,and the spirit,activity ability,stool shape and color were significantly improved.The growth trend of body weight in the drug treatment group was higher than that in the model group,but slower than that in the blank group,which was not statistically significant.In Zhuangganzhuyu Decoction group,the liver edge became sharp,the swelling of liver body was reduced,there was no obvious granule,the surface gloss was acceptable,the texture was soft,the appearance color was similar,close to reddish brown.In colchicine group,the liver particles decreased,the edge was blunt,and the color was yellowish white.2.Effects of Zhuanggan Zhuyu Decoction on serum ALT,AST,ALB and ALP levels in rats with hepatic fibrosis Compared with model group,the serum levels of ALT and AST in zhuanggan Zhuyu Decoction high-dose group decreased,and the serum levels of ALB increased(P<0.01);the serum levels of ALT and ALP in zhuanggan Zhuyu Decoction medium-dose and low-dose groups decreased(P<0.01,P<0.05),and the serum levels of ALB increased(P<0.01).The serum ALT level in colchicine group was decreased(P<0.05).3.Effect of zhuanggan Zhuyu Decoction on the morphological changes of liver fibrosis rats The results of HE and Masson showed that compared with the model group,the number of necrosis and deformation of liver cells in zhuanggan Zhuyu Decoction treatment group was reduced,the proliferation of fibrous tissue and the area of fibrous septum was reduced,the infiltration degree of inflammatory cells was reduced,and no false lobule formation was observed.Colchicine group had a certain degree of improvement in fibrous septal area.4.Effect of Zhuanggan Zhuyu Decoction on expression of MST1,YAP and LATS1 in hepatic fibrosis rats(1)WB result: Compared with blank group,the expression of YAP protein in model group increased(P<0.05),LATS1 protein expression decreased(P<0.01),and MST1 protein expression decreased,with no statistical significance.Compared with the model group,the expression of LATS1 protein in liver tissue of high-dose zhuanggan Zhuyu Decoction group was significantly higher(P<0.01),The expression of YAP protein decreased and the expression of LATS1 protein increased in the middle dose group of zhuanggan Zhuyu Decoction(P<0.01,P<0.05),The expression of Mst1 protein in liver tissue increased in the low dose group of Zhuanggan zhuyu Decoction(P<0.01),In colchicine group,the expression of MST1 protein was increased(P<0.05),and the expression of YAP protein was decreased(P<0.01).(2)PCR results: Compared with the blank group,the expression of MST1 m RNA in the liver tissue of the model group decreased,but the difference was not statistically significant.The expression of LATS1 m RNA decreased,and the expression of YAPm RNA increased(P<0.01).Compared with model group,the expression of MST1 and LATS1 m RNA in zhuanggan zhuyujian high-dose,medium-dose and low-dose groups and colchicine group increased,and the expression of YAPm RNA decreased(P<0.01).Conclusion:1.Zhuanggan Zhuyu Decoction can obviously improve the liver function of liver fibrosis rats,reduce the degree of liver fibrosis in liver tissue,and has a good liver protection effect.2.The mechanism of zhuang Gan Zhuyu Jian’s anti-liver fibrosis may be through up-regulating the expression of MST1 and LAST1 and down-regulating the expression of YAP.