Effect Of Estradiol On Apoptosis Of Hepatic Stellate Cells Through GPR30

Objectives To investigate the effect and mechanism of estradiol on apoptosis of Hepatic stellate cell through G protein coupled receptor 30（GPR30）,which can provide theoretical basis and new ideas for preventing or reversing liver fibrosis.Methods 1 Clinical research:A total of 179 patients with mild,moderate,severe viral hepatitis B and hepatitis B cirrhosis were selected from Tangshan Infectious Disease Hospital from September to December 2020,and during the same period,88 patients without hepatitis B were selected.General demographic characteristics and laboratory indicators（liver function,liver fiber and HBV-DNA）of the subjects were collected,and4m L fasting venous blood was collected from the subjects.The expressions of E₂and cytochrome C（Cyt C）and m RNA expressions of GPR30 and caspase-9 in the blood of the subjects were detected by ELISA and RT-q PCR.X²test was used to analyze the age and gender differences of patients with different lesions,and Pearson analysis was used to analyze the correlation between indicators.2 Experimental research:Resuscated human HSC stored in liquid nitrogen and divided the cells into 5 groups after good cell growth and synchronization:Control group,TGF-β1 group,TGF-β1+E₂（E₂）group,TGF-β1+GPR30 inhibitor（G-36）group,TGF-β1+E₂+GPR30 inhibitor（E₂+G-36）group.After the cells were grouped into the corresponding groups described above.Cell proliferation rate was detected by CCK-8.Apoptosis rate was detected by Annexin V-FITC/PI double staining,and protein expression levels of GPR30,Cyt C,caspase-9,caspase-3,Bcl-2 and Bax were determined by Western blot.Results 1 Clinical research:With the aggravation of hepatitis B disease,the four indexes of blood liver function,liver fiber and HBV-DNA load increased（P<0.05）,while the blood liver function,liver fiber and HBV-DNA load in patients with cirrhosis were slightly decreased compared with those in patients with severe hepatitis B（P<0.05）.The levels of E₂,Cyt C,GPR30 and caspase-9 decreased with the severity of hepatitis B disease（P<0.05）.Correlation analysis showed that liver function index was significantly positively correlated with HBV-DNA load and liver fiber（P<0.05）;E₂,Cyt C,GPR30and caspase-9 were negatively correlated with liver fibrosis（P<0.05）;E₂,GPR30 and caspase-9 were negatively correlated with ALT,AST and HBV-DNA（P<0.05）;Cyt C was negatively correlated with ALT and HBV-DNA（P<0.05）.2 Experimental research:Morphological changes of HSC:HSC in the control group was irregular,with large intercellular space and clear background;compared with the control group,cells in the TGF-β1 group was increased,the shape was fusiform,and the space was reduced.Compared with the TGF-β1 group,cells in the E₂group was significantly decreased,the cell volume was reduced and the process was reduced,the background was cloudy,and the cell space was increased.Compared with the TGF-β1 group,cells in the E₂+G-36group was significantly reduced,and the cells became shorter,fewer processes,and the intercellular space was significantly increased with unclear background.HSC proliferation:Compared with the control group,the relative growth rate（RGR）of TGF-β1and G-36 groups was significantly increased（P<0.05）;Compared with TGF-β1 group,RGR in E₂group and E₂+G-36 group was significantly decreased（P<0.05）;Compared with TGF-β1 group,RGR in G-36 group was significantly increased（P<0.05）;Compared with E₂group,RGR in G-36 group and E₂+G-36 group was significantly increased（P<0.05）;RGR in E₂+G-36 group was significantly lower than that in G-36group（P<0.05）.HSC apoptosis:Compared with TGF-β1 group,the apoptosis rate of E₂group and E₂+G-36 group was significantly increased（P<0.05）,but decreased significantly in G-36 group（P<0.05）.Compared with E₂group,the apoptosis rate of G-36 group and E₂+G-36 group decreased significantly（P<0.05）.Compared with G-36group,the apoptosis rate of E₂+G-36 group was significantly increased（P<0.05）.Western Blot showed that compared with the control group and TGF-β1 group,the expression levels of GPR30,Cyt C,caspase-9,caspase-3 and Bax were significantly increased in E₂group and E₂+G-36 group（P<0.05）,but significantly decreased in G-36group（P<0.05）;Compared with E₂group,the expression levels of GPR30,Cyt C,caspase-9,caspase-3 and Bax decreased significantly in G-36 group and E₂+G-36 group（P<0.05）;Compared with G-36 group,the expression levels of GPR30,Cyt C,caspase-9,caspase-3 and Bax were significantly increased in E₂+G-36 group（P<0.05）.The expression level of Bcl-2 in each group was opposite to that of Bax.Conclusions 1 With the aggravation of hepatitis B disease,the more severe the liver damage,the higher the liver fibrosis index,the more active HBV-DNA replication,the blood E₂and Cyt C,GPR30,caspase-9 level decreased.2 E₂can induce HSC apoptosis through its receptor GPR30,which may be caused by mitochondrial apoptosis pathway.Figure16;Table20;Reference138...