青蒿琥酯调节HMGB1-RAGE信号通路对缺氧复氧诱导的HK-2细胞损伤的影响  

Effects of artesunate on hypoxia reoxygenation-induced HK-2 cell injury by regulating HMGB1-RAGE signaling pathway

作　　者：陈娜 李兰梅 白伟伟 孙绍婷 王萌 张明 李亚芬 CHEN Na;LI Lanmei;BAI Weiwei;SUN Shaoting;WANG Meng;ZHANG Ming;LI Yafen(Department of Nephrology,Cangzhou People's Hospital,Cangzhou 061000,China;Medical Record Room,Cangzhou Integrated Traditional Chinese and Western Medicine Hospital,Cangzhou 061000,China)

机构地区：[1]沧州市人民医院肾病内科,061000 [2]沧州中西医结合医院病案室,061000

出　　处：《免疫学杂志》2024年第11期812-817,共6页Immunological Journal

基　　金：河北省2024年度医学科学研究课题(20240478)。

摘　　要：目的探讨青蒿琥酯(ART)调节高迁移率族蛋白B1(HMGB1)-晚期糖基化终产物受体(RAGE)信号通路对缺氧复氧(H/R)诱导的人肾近曲小管细胞HK-2细胞损伤的影响。方法将体外培养的HK-2细胞随机分为Control组、Model组、L-ART(4μg/ml ART)、H-ART组(16μg/ml ART)、H-ART+pcDNA-NC组(16μg/ml ART+转染pcDNA-NC质粒)、H-ART+pcDNA-HMGB1组(16μg/ml ART+转染pcDNA-HMGB1质粒)。除Control组正常培养,其余各组建立H/R细胞损伤模型。MTT和平板克隆形成实验检测各组细胞的增殖;流式细胞仪检测各组细胞的凋亡;ELISA试剂盒检测各组细胞中白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)的表达;Western blot检测各组细胞中增殖相关蛋白(PCNA)、凋亡相关蛋白(Bcl-2、Bax)及HMGB1-RAGE信号通路相关蛋白(HMGB1、RAGE)表达情况。结果Model组与Control组相比,HK-2细胞的存活率、克隆数、PCNA、Bcl-2蛋白表达降低,但细胞凋亡率、IL-1β、IL-6、TNF-α、Bax、HMGB1、RAGE蛋白表达升高(P<0.05);L-ART组、H-ART组与Model组相比,细胞的存活率、克隆数、PCNA、Bcl-2蛋白表达升高,但细胞凋亡率、IL-1β、IL-6、TNF-α、Bax、HMGB1、RAGE蛋白表达降低(P<0.05);H-ART+pcDNA-HMGB1组与H-ART、H-ART+pcDNA-NC组相比,细胞的存活率、克隆数、PCNA、Bcl-2蛋白表达降低,但细胞凋亡率、IL-1β、IL-6、TNF-α、Bax、HMGB1、RAGE蛋白表达升高(P<0.05)。结论ART可能是通过抑制HMGB1-RAGE信号通路,抑制炎症反应及细胞凋亡,促进细胞增殖,减轻H/R诱导的肾近曲小管细胞HK-2细胞损伤。Objective To investigate the effect of artesunate(ART)on hypoxia/reoxygenation(H/R)-induced injury of human renal proximal tubular HK-2 cells by regulating the high mobility group protein B1(HMGB1)-receptor of advanced glycation endproduct(RAGE)signaling pathway.Methods HK-2 cells cultured in vitro were stochastically separated into control group,model group,L-ART group(4μg/ml ART),H-ART group(16μg/ml ART),H-ART+pcDNA NC group(16μg/ml ART+transfected pcDNA NC plasmid),and H-ART+pcDNA-HMGB1 group(16μg/ml ART+transfected pcDNA-HMGB1 plasmid).HK-2 cells in the control group were cultured normally,while the cells in the other groups were induced with hypoxia/reoxygenation(H/R).MTT and plate clone formation experiments were applied to detect the proliferation of cells in each group;ELISA kits were used to measure the expression of interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)in various groups;flow cytometry was applied to detect the apoptosis in various groups;Western blot was applied to detect the expression of proliferation related proteins(PCNA),apoptosis related proteins(Bcl-2,Bax),and HMGB1-RAGE signaling pathway related proteins(HMGB1,RAGE)in each group.Results Compared with the control group,the survival rate,clone number,PCNA and Bcl-2 proteins expression of HK-2 cells decreased in the model group,but the apoptosis rate,IL-1β,IL-6,TNF-α,Bax,HMGB1 and RAGE proteins expression increased(P<0.05).Compared with the model group,the survival rate,clone number,PCNA and Bcl-2 proteins expression of HK-2 cells increased in L-ART and H-ART groups,but the apoptosis rate,IL-1β,IL-6,TNF-α,Bax,HMGB1 and RAGE proteins expression decreased(P<0.05).Compared with the H-ART and H-ART+pcDNA NC groups,the survival rate,clone number,PCNA and Bcl-2 proteins expression of HK-2 cells decreased in the H-ART+pcDNA-HMBB1 group,but the apoptosis rate,IL-1β,IL-6,TNF-α,Bax,HMGB1 and RAGE proteins expression increased(P<0.05).Conclusion ART may suppress inflammatory response and cell apoptosis,promote cell proliferat

关 键 词：青蒿琥酯 高迁移率族蛋白B1-晚期糖基化终产物受体信号通路 缺氧复氧 人肾近曲小管细胞 细胞损伤 

分 类 号：R692.6[医药卫生—泌尿科学]