利拉鲁肽抗CXC趋化因子配体16诱导的人足细胞脂质沉积作用及机制  

Effect and mechanism of liraglutide against CXC-chemokine ligand 16-induced lipid deposition in human podocytes

作　　者：陈瑛 曹爱丽[1] CHEN Ying;CAO Aii(Department of Nephrology,Putuo Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China)

机构地区：[1]上海中医药大学附属普陀医院肾内科,上海200062

出　　处：《中国药理学与毒理学杂志》2023年第11期823-832,共10页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(81400728);上海市浦江人才计划(21PJ1412400);上海市普陀区卫生健康系统科技创新项目(ptkwws202001);上海中医药大学预算内项目(2020LK070)。

摘　　要：目的探讨利拉鲁肽(liraglutide)抗CXC趋化因子配体16(CXCL16)诱导的人足细胞脂质沉积的作用及机制。方法①葡萄糖40 mmol·L^(-1)刺激人足细胞24 h,棕榈酸250μmol·L^(-1)刺激人足细胞6 h,Western印迹法检测足细胞CXCL16蛋白表达水平。②重组CXCL16100μg·L^(-1)刺激足细胞0,6,12和24 h,Western印迹法检测足细胞裂隙素蛋白表达水平。③足细胞分为细胞对照组、CXCL16100μg·L^(-1)组、CXCL16+利拉鲁肽10,50和100 nmol·L^(-1)组及CXCL16+辛伐他汀100 nmol·L^(-1)组,加药2 h后加重组CXCL16100μg·L^(-1)继续培养24 h,油红O染色检测足细胞脂滴面积。④足细胞分为细胞对照组、CXCL16100μg·L^(-1)组、CXCL16+利拉鲁肽100 nmol·L^(-1)组和CXCL16+辛伐他汀100 nmol·L^(-1)组,加药2 h后加CXCL16100μg·L^(-1)继续培养24 h,鬼笔环肽染色检测足细胞肌动蛋白应力纤维百分比,ELISA检验细胞培养液中肿瘤坏死因子α(TNF-α)、转化生长因子β(TGF-β)和白细胞介素1β(IL^(-1)β)蛋白浓度,实时荧光定量PCR检测TNF-α,TGF-β和IL^(-1)βmRNA表达水平;Western印迹法检测足细胞裂隙素、胆固醇调节元件结合蛋白1(SREBP1)、SREBP2和脂肪分化相关蛋白(ADRP)表达水平。结果①与细胞对照组相比,高糖和棕榈酸刺激均可引起人足细胞CXCL16表达水平显著升高(P<0.01)。②与细胞对照组相比,CXCL16刺激可引起足细胞裂隙素表达显著下调(P<0.01)。③与细胞对照组相比,CXCL16刺激可引起足细胞脂滴面积显著升高(P<0.01),辛伐他汀和不同浓度利拉鲁肽均可显著缓解这一变化(P<0.01)。④与细胞对照组相比,重组CXCL16可引起足细胞肌动蛋白应力纤维百分比显著下降(P<0.01),培养液中TNF-α,TGF-β和L^(-1)β蛋白浓度显著升高(P<0.01),足细胞TNF-α,TGF-β和IL^(-1)βmRNA水平显著升高(P<0.05,P<0.01),裂隙素表达显著下调(P<0.01),SREBP1,SREBP2和ADRP蛋白表达水平显著增高(P<0.01);与CXCL16组比较,利拉鲁肽和辛伐他汀可OBJECTIVE To explore the effect and mechanism of liraglutide on CXC-chemokine ligand 16(CXCL16)-induced lipid deposition in human podocytes.METHODS①Human podocytes were stimulated with high glucose(HG)40 mmol·L^(-1) for 24 h and palmitic acid(PA)250μmol·L^(-1) for 6 h.Western blotting was used to detect the protein expression level of CXCL16 in podocytes.②CXCL16 was used to stimulate podocytes at a concentration of 100μg·L^(-1) for 0,6,12,and 24 h.Western blotting was used to detect the protein expression level of nephrin in podocytes.③Podocytes were divided into the cell control group,CXCL1610μg·L^(-1),CXCL16+liraglutide 10,50,100 nmol·L^(-1) group and CXCL16+simvastatin 100 nmol·L^(-1) group.After two hours of drug incubation,CXCL16100μg·L^(-1) was added and cultured for 24 h.Oil Red O staining was used to detect the lipid droplet area in podocytes.④Podo-cytes were divided into the cell control group,CXCL1610μg·L^(-1) gruop,CXCL16+liraglutide 100 nmol·L^(-1) group and CXCL16+simvastatin 100 nmol·L^(-1) group.After two hours of incubation,CXCL16 was added and cultured for 24 h.Phalloidin staining was used to detect podocyte stress fibers.ELISA was used to measure the concentrations of tumor necrosis factorα(TNF-α),transforming growth factor-β(TGF-β)and interleukin-1β(IL^(-1)β)in cell culture supernatants.Real-time fluorescence quantitative PCR was used to detect the mRNA expression levels of TNF-α,TGF-β,and IL^(-1)β.Western blotting was used to detect the protein expression levels of nephrin,sterol regulatory element-binding protein 1(SREBP1),SREBP2,and adipose differentiation-related protein(ADRP)in podocytes.RESULTS①Compared with the cell control group,both high glucose and palmitic acid stimulation significantly increased CXCL16 expression levels in human podocytes(P<0.01).②Compared with the cell control group,recombinant CXCL16 stim-ulation significantly down-regulated the expression of nephrin in podocytes(P<0.01).③Compared with the cell control group,recombinant CXCL16

关 键 词：利拉鲁肽 足细胞损伤 脂质沉积 CXC趋化因子配体16 

分 类 号：R963[医药卫生—微生物与生化药学] R977[医药卫生—药理学]