卡格列净对高糖环境下小鼠肾小球系膜细胞的抗炎、抗凋亡作用及其机制  

Anti-inflammation and anti-apoptosis effects of canagliflozin on glomerular mesangial cells under high glucose

作　　者：尹一帆 谭睿陟 王丽 刘建 YIN Yifan;TAN Ruizhi;WANG Li;LIU Jian(Department of Nephrology,The Third People’s Hospital of Chengdu,Affiliated Hospital of Southwest Jiaotong University,Chengdu 610014,China;不详)

机构地区：[1]成都市第三人民医院西南交通大学附属医院肾内科,成都610014 [2]西南医科大学附属中医医院中西医结合研究中心 [3]西南医科大学附属中医医院肾内科

出　　处：《山东医药》2022年第32期16-20,共5页Shandong Medical Journal

基　　金：四川省科学技术厅与泸州市人民政府、泸州医学院联合科研专项资金计划项目(LY-448);四川省科学技术厅项目(LY-448)。

摘　　要：目的观察卡格列净对高糖环境下小鼠肾小球系膜细胞SV40 MES13的抗炎、抗凋亡作用,并探讨其机制。方法将SV40 MES13细胞随机分为正常组、高糖组、卡格列净1组、卡格列净2组;正常组在低糖培养基中培养,高糖组、卡格列净1组、卡格列净2组在高糖培养基中培养,卡格列净1组、卡格列净2组分别加入5、15μoml/L的卡格列净;培养24 h后,采用ELISA法检测各组培养液上清中的单核细胞趋化蛋白1(MCP-1)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6),免疫荧光法检测细胞中的TNF-α、IL-6,流式细胞仪测算细胞凋亡率,Western blotting法检测细胞中的转化生长因子β_(1)(TGF-β_(1))、p-Smad3、Smad3。将SV40 MES13细胞分为NC组、HG组、HG+Can组、HG+Can+Smad3 OE组;NC组在低糖培养基中培养,HG组、HG+Can组、HG+Can+Smad3 OE组在高糖培养基中培养,HG+Can组加入15μoml/L卡格列净,HG+Can+Smad3 OE组转染pcDNA3.1-Smad3质粒24 h后加入15μoml/L卡格列净;培养24 h后,采用Western blotting法检测各组细胞中的TGF-β_(1)、p-Smad3、Smad3,免疫荧光法检测细胞中的TNF-α、IL-6,流式细胞仪测算细胞凋亡率。结果高糖组上清液MCP-1、TNF-α、IL-6水平及细胞中TNF-α、IL-6表达高于正常组,卡格列净1组、卡格列净2组上述指标水平低于高糖组,且卡格列净2组低于卡格列净1组(P均<0.05);高糖组早、晚期凋亡细胞比例及TGF-β_(1)表达、p-Smad3/Smad3高于正常组,卡格列净1组、卡格列净2组早期凋亡细胞比例及TGF-β_(1)表达、p-Smad3/Smad3低于高糖组,卡格列净2组早期凋亡细胞比例低于卡格列净1组(P均<0.05)。HG组TGF-β_(1)、p-Smad3/Smad3、TNF-α、IL-6表达及早、晚期凋亡细胞比例高于NC组,HG+Can组上述指标低于HG组,HG+Can+Smad3 OE组TGF-β_(1)、p-Smad3/Smad3、TNF-α表达及早、晚期凋亡细胞比例高于HG+Can组(P均<0.05)。结论卡格列净可抑制高糖环境下肾小球系膜细胞炎症因子分泌及细胞凋�Objective To observe the anti-inflammation and anti-apoptosis effects of canagliflozin on high glucoseinduced glomerular mesangial cells,and to explore its possible mechanisms.Methods SV40 MES13 cells were randomly divided into normal control group,high glucose group,HG+canagliflozin 5μmol/L group(canagliflozin 1 group),and HG+canagliflozin 15μmol/L group(canagliflozin 2 group).The cells in the normal control group were cultured in 5.5 mmol/L DMEM medium,and cells in the other groups were cultured in 35 mmol/L DMEM medium;5μmol/L canagliflozin was added to the canagliflozin 1 group,and 15μmol/L canagliflozin was added to the canagliflozin 2 group.All groups were cultured for 24 h,the levels of monocyte chemoattractant protein(MCP)-1,tumor necrosis factor(TNF)-α,and interleukin(IL)-6 were detected by ELISA;the levels of TNF-αand IL-6 were detected by immunofluorescence assay;the apoptotic rate was measured by flow cytomtry,and the expression levels of transforming growth factor(TGF)-β_(1),p-Smad3,and Smad3 were detected by Western blotting.SV40 MES13 cells were randomly divided into NC group,HG group,HG+Can group,and HG+Can+Smad3 OE group.The cells in the NC group were cultured in 5.5 mmol/L DMEM medium,and cells in the other groups were cultured in 35mmol/L DMEM medium;15μmol/L canagliflozin was added to the HG+Can group;pcDNA3.1-Smad3 plasmid was transfected into the HG+Can+Smad3 OE group,and 15μmol/L canagliflozin was added at 24 h after transfection.All groups were cultured for 24 h,the expression levels of TGF-β_(1),p-Smad3,and Smad3 were detected by Western blotting,the levels of TNF-αand IL-6 were detected by immunofluorescence assay,and the apoptotic rate was measured by flow cytomtry.Results The levels of MCP-1,TNF-α,and IL-6 in supernatant and the expression levels of TNF-αand IL-6 in cells in the high glucose group were higher than those in the normal control group,the levels of the above indexs in the canagliflozin 1 group and canagliflozin 2 group were lower than those in the high glucose

关 键 词：卡格列净 糖尿病 肾小球系膜细胞 炎症反应 细胞凋亡 TGF-β/Smad3信号通路 

分 类 号：R587.2[医药卫生—内分泌]