二甲双胍对草酸钙诱导的人肾小管上皮细胞HK-2氧化应激及炎症反应调节作用观察  

Regulatory effects of metformin on oxidative stress and inflammatory response of human renal tubular epithelial cells HK-2 induced by calcium oxalate

作　　者：周吾溪 王海荣[1] 徐友胜 戚剑烽 陆顶进 ZHOU Wuxi;WANG Hairong;XU Yousheng;QI Jianfeng;LU Dingjin(Department of Urology,Beihai People's Hospital,Beihai 536000,China)

机构地区：[1]北海市人民医院泌尿外科,广西北海536000

出　　处：《山东医药》2023年第9期26-30,共5页Shandong Medical Journal

基　　金：广西医科大学青年科学基金资助项目(GXMUYSF202138)。

摘　　要：目的观察二甲双胍对草酸钙诱导的人肾小管上皮细胞HK-2氧化应激及炎症反应的调节作用。方法将HK-2细胞随机分成对照组、草酸钙组、二甲双胍组1、二甲双胍组2、PDTC组,其中对照组给予无血清DMEM培养基,草酸钙组加入浓度为100μg/mL的草酸钙晶体悬浮液,二甲双胍组1加入浓度为100μg/mL的草酸钙晶体悬浮液和1.20 mmol/L的二甲双胍晶体悬浮液,二甲双胍组2加入浓度为100μg/mL的草酸钙晶体悬浮液和0.80 mmol/L的二甲双胍晶体悬浮液,PDTC组加入浓度为100μg/mL的草酸钙晶体悬浮液和100 mmol/L的NF-κB特异性抑制剂PDTC晶体悬浮液。各组细胞继续培养6 h,采用比色法检测细胞上清液中丙二醛(MDA),采用Elisa法检测细胞上清液中单核细胞趋化蛋白-1(MCP-1)蛋白,采用RT-PCR法检测细胞中NF-κB、MCP-1 mRNA。结果对照组、草酸钙组、二甲双胍组1、二甲双胍组2、PDTC组细胞上清液中MDA含量分别为(0.21±0.02)、(1.62±0.09)、(1.04±0.01)、(1.27±0.12)、(0.83±0.02)mmol/mg,MCP-1蛋白含量分别为(8.25±0.62)、(66.31±3.78)、(31.54±2.15)、(42.26±2.10)、(20.36±1.92)pg/mL,组间相比,P均<0.05。对照组、草酸钙组、二甲双胍组1、二甲双胍组2、PDTC组细胞中NF-κB mRNA相对表达量分别为1.00±0.01、1.68±0.02、1.23±0.02、1.41±0.01、1.09±0.01,MCP-1 mRNA相对表达量分别为1.00±0.01、2.39±0.01、1.64±0.02、1.89±0.01、1.44±0.01,各组细胞中NF-κB、MCP-1 mRNA相对表达量组间相比,P均<0.05。结论1.20 mmol/L或0.80 mmol/L二甲双胍可抑制草酸钙诱导的HK-2细胞氧化应激,并通过抑制NF-κB通路进一步抑制下游炎症反应。Objective To observe regulatory effects of metformin on oxidative stress and inflammatory response of human renal tubular epithelial cells HK-2 induced by calcium oxalate.Methods HK-2 cells were randomly divided into five groups:the control group,calcium oxalate group,metformin group 1,metformin group 2,and PDTC group.Cells in the control group were cultured in serum-free DMEM medium,cells in the calcium oxalate group were added with 100μg/mL calcium oxalate crystal suspension,cells in the metformin group 1 were added with 100μg/mL calcium oxalate crystal suspension and 1.20 mmol/L metformin crystal suspension,cells in the metformin group 2 were added with 100μg/mL calcium oxalate crystal suspension and 0.80 mmol/L metformin crystal suspension,and cells in the PDTC group were added with 100μg/mL calcium oxalate crystal suspension and 100 mmol/L NF-κB specific inhibitor PDTC crystal suspension.The cells in each group continued to be cultured for 6 h;colorimetric method was used to detect malondialdehyde(MDA)in cell supernatant,ELISA was used to detect the monocyte chemotactic protein-1(MCP-1)protein in cell supernatant,and RT-PCR was used to detect the mRNAs of NF-κB and MCP-1.Results The content of MDA in the cell supernatant of the control group,calcium oxalate group,metformin group 1,metformin group 2,and PDTC group was(0.21±0.02),(1.62±0.09),(1.04±0.01),(1.27±0.12)and(0.83±0.02)mmol/mg,respectively;the protein content of MCP-1 was(8.25±0.62),(66.31±3.78),(31.54±2.15),(42.26±2.10),and(20.36±1.92)pg/mL,respectively;there were significant differences between groups(all P<0.05).The relative expression levels of NF-κB mRNA in the control group,calcium oxalate group,metformin group 1,metformin group 2,and PDTC group were 1.00±0.01,1.68±0.02,1.23±0.02,1.41±0.01,and 1.09±0.01,respectively;the relative expression levels of MCP-1 mRNA were 1.00±0.01,2.39±0.01,1.64±0.02,1.89±0.01,and 1.44±0.01,respectively;there were significant differences in the mRNA expression levels of NF-κB and MCP-1 betw

关 键 词：二甲双胍 肾结石 草酸钙肾结石 人肾小管上皮细胞 丙二醛 单核细胞趋化蛋白-1 

分 类 号：R749.16[医药卫生—神经病学与精神病学]