Mechanism Of Shenshuai Xiezhuo Decoction Against Ang-?-induced Glomerular Endothelial Cell Injury

Objectives:The in-vivo study of the instructor's previous topic"shenshuai Xiezhuo Decoction Regulates Microvascular Protection in Rats with Chronic Renal Failure"shows that Shenshuai Xiezhuo Decoction can reduce endothelial damage markers by regulating the expression of vascular endothelial growth factor?VEGF?receptor.The content of vWF can play a protective role in the damage of renal vascular endothelial cells in rats with chronic renal failure.In order to further study the protective mechanism of Shenshuai Xiezhuo Decoction on glomerular endothelial cells,this study explored the proliferation and apoptosis of angiotensin ?-stimulated glomerular endothelial cells?GECs?by Shenshuai Xiezhuo Decoction.Death status,expression of transforming growth factor?TGF-??and connective tissue growth factor?CTGF?protein,levels of ROS in cells,and changes in inflammatory mediators tumor necrosis factor?TNF-??and interleukin-6?IL-6?levels The regulation effect further clarifies the mechanism of Shenshuai Xiezhuo Decoction against Ang-?-induced glomerular endothelial cell injury,and provides a theoretical basis for the prevention of microvascular protection of glomerular endothelial cells by Shenshuai Xiezhuo Decoction.Methods:Rat glomerular endothelial cells were stimulated with 1×10^-6mol/L Ang? by in vitro experiments.According to the intervention situation,it was divided into the control group,the model group,the high-dose group of traditional Chinese medicine?the high-dose drug-containing serum intervention of Shenshuai Xiezhuo Decoction?,the middle dose group of traditional Chinese medicine?the mid-dose drug-containing serum intervention of Shenshuai Xiezhuo Decoction?,and the low dose group of traditional Chinese medicine?the low-dose drug-containing serum intervention of Shenshuai Xiezhuo Decoction?,Western medicine group?Kosuya drug-containing serum intervention?.MTT assay was used to detect cell proliferation,AV/PI flow cytometry was used to detect apoptosis,ELISA was used to detect TNF-?,IL-6,cytokine TGF-?and CTGF protein content,and QRT-PCR was used to detect TNF.The expression of?,IL-6,TGF-?and CTGF mRNA and the content of reactive oxygen species ROS were detected by fluorescent probe method.Results:1.The results of proliferation:After Ang? stimulation,when the concentration of drug-containing serum was 10%,the proliferation of cells:the proliferation rate of the model group was lower than that of the control group?P<0.01?;the low-dose group of traditional Chinese medicine,the middle-dose group of traditional Chinese medicine,the high-dose group of traditional Chinese medicine and The proliferation rate of the western medicine group was higher than that of the model group?P<0.05?.There was no significant difference between the proliferation rate of western medicine group and traditional Chinese medicine high dose group,Chinese medicine middle dose group and Chinese medicine low dose group?P>0.05?.2.The results of apoptosis:the apoptosis rate of the model group was higher than that of the control group?P<0.01?.The apoptosis rate of the low-dose group,the middle-medication group,the high-dose group and the western medicine group was lower than that of the model group.?P<0.05?,the apoptosis rate of Chinese medicine middle dose group and Chinese medicine high dose group was lower than that of Chinese medicine low dose group?P<0.05?.3.The results of ELISA:the levels of TNF-?,IL-6,TGF-?and CTGF in the model group were higher than those in the control group?P<0.01?.The low dose group of traditional Chinese medicine,the middle dose group of traditional Chinese medicine,the high dose group of traditional Chinese medicine and western medicine.The levels of TNF-?,IL-6,TGF-?and CTGF were decreased in the group compared with the model group?P<0.05?.The contents of TNF-?,IL-6,TGF-?and CTGF in the middle dose group and the high dose group of traditional Chinese medicine decreased compared with the low dose group?P<0.05?.4.The results of PCR:The expression of TNF-?,IL-6,TGF-?and CTGF mRNA in the model group was increased?P<0.01?.The low dose group of traditional Chinese medicine,the middle dose group of traditional Chinese medicine,the high dose group of traditional Chinese medicine and the western medicine group were TNF-?,IL-6 and TGF.The expression of?-and CTGF mRNA was lower than that of the model group?P<0.05?.The expressions of TNF-?,IL-6,TGF-?and CTGF mRNA in the middle dose group and the high dose group of traditional Chinese medicine were lower than those in the low dose group?P<0.05?.5.The results of ROS:After Ang? stimulation,the ROS content of reactive oxygen species in the model group increased?P<0.01?.The content of reactive oxygen species ROS in the low-dose group,the middle-medication group,the high-dose group and the western medicine group decreased compared with the model group.P<0.05).The content of reactive oxygen species ROS in the middle dose group and the high dose group of traditional Chinese medicine was lower than that in the low dose group?P<0.05?.Conclusions:1.Shenshuai Xiezhuo Decoction can improve the damage of angiotensin ?-stimulated glomerular endothelial cells,protect endothelial cells and promote cell proliferation.2.Shenshuai Xiezhuo Decoction can inhibit the apoptosis of glomerular endothelial cells and the release of inflammatory mediators mediated by angiotensin ?,reduce the content of reactive oxygen species ROS in cells,reduce oxidative stress,inhibit cytokine CTGF,The expression of TGF-?protein reduces the inhibition of proliferation of damaged endothelial cells.