外源性硫化氢对肾小球系膜细胞缺氧/复氧损伤的影响及其机制  

Effect of exogenous hydrogen sulfide on hypoxia/reoxygenation injuryin glomerular mesangial cells and its mechanism

作　　者：林真亭 田华 李鸿珠 LIN Zhen-ting;TIAN Hua;LI Hong-zhu(Dept of Basic Medicine,Xiamen Medical College,Xiamen 361023,China;Dept of Basic Education,School of Medicine,Xiamen University,Xiamen 361102,China)

机构地区：[1]厦门医学院基础医学部,福建厦门361023 [2]厦门大学医学院基础部,福建厦门361102

出　　处：《中国药理学通报》2024年第1期133-138,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81770486);福建省自然基金资助项目(No 2022J011399);厦门医学院校级科研计划项目(No K2023-42)。

摘　　要：目的探讨外源性硫化氢(hydrogen sulfide,H_(2)S)对肾小球系膜细胞缺氧/复氧(hypoxia/reoxygenation,H/R)损伤的影响,并阐明其相关作用机制。方法H/R诱导小鼠肾小球系膜细胞系(SV40MES13)建立细胞损伤模型。细胞增殖试剂盒(CCK8)检测细胞活力,荧光探针技术分别检测H_(2)S和活性氧(ROS)含量,生化试剂盒检测超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,Hoechst 33342染色检测细胞凋亡率;Western blot检测胱硫醚-γ-裂解酶(cystathione-gamma-lyase,CSE)、细胞凋亡相关蛋白(cleaved-caspase-3、Cyt c和Bcl-2)表达及信号通路相关蛋白(ERK1/2、Phospho-ERK1/2)活性。结果与对照(Control)组相比,缺氧/复氧(H/R)组细胞活力、内源性H_(2)S含量CSE蛋白表达、SOD活性及Bcl-2蛋白表达均明显降低;细胞凋亡率、MDA和ROS含量、Cyt c及cleaved caspase-3蛋白表达均显著升高;同时,磷酸化ERK1/2(p-ERK1/2)活性明显降低。与H/R比较,H/R+NaHS(外源性H_(2)S供体)逆转了H/R对上述指标的影响。另外,PD98059(ERK1/2抑制剂)减弱了NaHS对磷酸化ERK1/2的作用。结论肾小球系膜细胞H/R损伤与内源性CSE/H_(2)S系统下调有关;外源性H_(2)S通过上调ERK1/2通路抑制氧化应激和细胞凋亡,减轻肾小球系膜细胞H/R损伤。Aim To explore the effect of exogenous hydrogen sulfide(H_(2)S)on hypoxia/reoxygenation(H/R)injury in glomerular mesangial cells and elucidate its relevant mechanism.Methods H/R induced mouse mesangial cell line(SV40MES13)to establish cell damage model.Cell viability was detected by cell proliferation kit(CCK8),the content of H_(2)S and reactive oxygen species(ROS)was detected by fluorescence probe technology,superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were observed through biochemical kit,the apoptosis rate was tested via Hoechst 33342 staining,the expression of cystathione-gamma-lyase(CSE)and apoptosis-related proteins(cleaved caspase-3,Cyt c and Bcl-2)as well as activity of signaling pathway-related proteins(ERK1/2,phospho-ERK1/2)were analyzed by Western blotting.Results Compared with the Control group,cell viability,endogenous H_(2)S content and CSE protein expressions,SOD activity and Bcl-2 protein expressions significantly decreased in hypoxia/reoxygenation(H/R)group;the apoptosis rate,the content of MDA and ROS,the expression of Cytc and Cleaved caspase-3 protein were significantly increased;In addition,phosphorylation of ERK1/2(p-ERK1/2)was significantly decreased.Compared with H/R,D-gal+NaHS(an exogenous H_(2)S donor)reversed the effect of H/R on the above indexes.In addition,PD98059(an ERK1/2 inhibitor)cancelled the effect of NaHS on p-ERK1/2.Conclusions The H/R injury of glomerular mesangial cells is related to the down-regulation of endogenous CSE/H_(2)S system,and exogenous H_(2)S inhibits oxidative stress and apoptosis by up-regulating ERK1/2 pathway,thereby alleviating the H/R injury of glomerular mesangial cells.

关 键 词：硫化氢 缺氧/复氧 肾小球系膜细胞 ERK1/2通路 细胞凋亡 CSE/H_(2)S系统 

分 类 号：R-332[医药卫生] R322.61R329.25R845.22R916.3