甘草素调节Hippo/YAP/TAZ信号通路对高糖诱导的肾小球系膜细胞炎症反应及纤维化的影响  

Impacts of liquiritigenin on high glucose-induced inflammatory response and fibrosis in glomerular mesangial cells by regulating Hippo/YAP/TAZ signaling pathway

作　　者：温玉[1] 付娆 刘城[1] 邱友春 WEN Yu;FU Rao;LIU Cheng;QIU Youchun(Department of Nephrology,Ziyang People's Hospital,Ziyang 641301,Sichuan,China;Department of Clinical Medicine,Southwest Medical University,Luzhou 646099,Sichuan,China)

机构地区：[1]资阳市人民医院肾脏内科,四川资阳641301 [2]西南医科大学临床医学系,四川泸州646099

出　　处：《西部医学》2024年第8期1131-1136,共6页Medical Journal of West China

基　　金：资阳市科技成果转化示范项目(Zykjjsc20-cgzh-2019-01)。

摘　　要：目的探讨甘草素(LQ)调节Hippo/Yes相关蛋白(YAP)/含有PDZ结合位点转录共激活因子(TAZ)通路对高糖(HG)诱导的肾小球系膜细胞炎症反应及纤维化的影响。方法将HBZY-1细胞分为对照组(Ct组)、高糖组(HG组)、低剂量LQ组(LQ-L组,20μmmol/L)、高剂量LQ组(LQ-H组,40μmmol/L)、pcDNA3.1组(转染pcDNA3.1)、pcDNA3.1-YAP/TAZ组(转染pcDNA3.1-YAP/TAZ)、LQ-H+pcDNA3.1组(40μmmol/L+转染pcDNA3.1)、LQ-H+pcDNA3.1-YAP/TAZ组(40μmmol/L+转染pcDNA3.1-YAP/TAZ)。qRT-PCR、Western blot检测转染效果。CCK-8法检测细胞增殖;ELISA检测细胞上清中TNF-α、单核细胞趋化蛋白-1(MCP-1)、IL-1β水平;免疫荧光检测IV型胶原纤维(Collagen IV)、纤连蛋白(FN)蛋白表达;Western blot检测结缔组织生长因子(CTGF)、TGF-β1、YAP1、TAZ蛋白表达。结果HBZY-1细胞成功高表达YAP1/TAZ;与Ct组比较,HG组HBZY-1细胞OD450值、TNF-α、MCP-1、IL-1β、Collagen IV、FN蛋白相对荧光强度、CTGF、TGF-β1、YAP1、TAZ蛋白表达升高(P<0.05);与HG组比较,LQ-L组、LQ-H组HBZY-1细胞OD450值、TNF-α、MCP-1、IL-1β、Collagen IV、FN蛋白相对荧光强度、CTGF、TGF-β1、YAP1、TAZ蛋白表达降低(P<0.05);与HG组、pcDNA3.1组比较,pcDNA3.1-YAP/TAZ组HBZY-1细胞OD450值、TNF-α、MCP-1、IL-1β、Collagen IV、FN蛋白相对荧光强度、CTGF、TGF-β1、YAP1、TAZ蛋白表达升高(P<0.05);pcDNA3.1-YAP/TAZ减弱了高剂量LQ对HG诱导的HBZY-1细胞炎症反应及纤维化的抑制作用。结论LQ可能通过抑制Hippo/YAP/TAZ通路抑制HG诱导的HBZY-1细胞炎症反应及纤维化。Objective To investigate the impacts of liquiritigenin(LQ)on high glucose(HG)-induced inflammatory response and fibrosis in glomerular mesangial cells by regulating Hippo/Yes-associated protein(YAP)/transcriptional coactivator with PDZ-binding motif(TAZ)pathway.Methods HBZY-1 cells were grouped into control group(Ct group),high glucose group(HG group),low-dose LQ group(LQ-L group,20μmmol/L),high-dose LQ group(LQ-H group,40μmmol/L),pcDNA3.1 group(transfected pcDNA3.1),pcDNA3.1-YAP/TAZ group(transfected pcDNA3.1-YAP/TAZ),LQ-H+pcDNA3.1 group(40μmmol/L+transfected pcDNA3.1),and LQ-H+pcDNA3.1-YAP/TAZ group(40μmmol/L+transfected pcDNA3.1-YAP/TAZ).The transfection effect was detected by qRT-PCR and western blot.Cell proliferation was detected by CCK-8 assay.The levels of tumor necrosis factor-α(TNF-α),monocyte chemoattractant protein-1(MCP-1)and interleukin-1β(IL-1β)in the cell supernatant were measured by ELISA.The protein expressions of collagen IV(Collagen IV)and fibronectin(FN)were measured by immunofluorescence.The protein expressions of connective tissue growth factor(CTGF),transforming growth factor-β1(TGF-β1),YAP1 and TAZ were measured by western blot.Results HBZY-1 cells successfully overexpressed YAP1/TAZ.Compared with the Ct group,the OD450 value,TNF-α,MCP-1,IL-1β,Collagen IV,FN protein relative fluorescence intensity,CTGF,TGF-β1,YAP1,TAZ protein expressions in HBZY-1 cells in HG group increased(P<0.05).Compared with the HG group,the OD450 value,TNF-α,MCP-1,IL-1β,Collagen IV,FN protein relative fluorescence intensity,CTGF,TGF-β1,YAP1,TAZ protein expression decreased in LQ-L group and LQ-H group(P<0.05).Compared with HG group and pcDNA3.1 group,the OD450 value,TNF-α,MCP-1,IL-1β,Collagen IV,FN protein relative fluorescence intensity,CTGF,TGF-β1,YAP1,TAZ protein expression in pcDNA3.1-YAP/TAZ group increased(P<0.05).pcDNA3.1-YAP/TAZ attenuated the inhibitory effects of high-dose LQ on HG-induced inflammatory response and fibrosis in HBZY-1 cells.Conclusion LQ may inhibit the HG-induced inflamm

关 键 词：甘草素 Hippo/Yes相关蛋白/含有PDZ结合位点转录共激活因子通路 肾小球系膜细胞 炎症 纤维化 

分 类 号：R692.6[医药卫生—泌尿科学]