三七皂苷R1对碘美普尔400诱导肾小管上皮细胞损伤的影响  

Effect of notoginsenoside R1 on renal tubular epithelial cell injury induced by Iomeprol 400

作　　者：龙芳敏 吕梁 LONG Fangmin;LYU Liang(The Affiliated Hospital of Kunming University of Science and Technology(The First People’s Hospital of Yunnan Province),Kunming 650032,Yunnan,China;School of Basic Medical Sciences,Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China)

机构地区：[1]昆明理工大学附属医院(云南省第一人民医院),云南昆明650032 [2]右江民族医学院基础医学院,广西百色533000

出　　处：《现代中西医结合杂志》2023年第10期1359-1365,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：云南省万人计划名医专项(YNWR-MY-2019-011);云南省卫生科技计划资助项目(2018NS0269)。

摘　　要：目的探讨三七皂苷R1对碘美普尔400诱导大鼠肾小管上皮细胞损伤的影响及其调控机制。方法NRK-52E细胞分为5组:正常对照组正常培养;三七皂苷组加入三七皂苷R1培养;碘美普尔组加入碘美普尔400培养;三七皂苷+碘美普尔组先加入三七皂苷R1培养,然后加入碘美普尔400培养;锌原卟啉组先加入锌原卟啉和三七皂苷R1培养,然后加入碘美普尔400培养。CCK-8法评估细胞活力筛选三七皂苷R1最佳保护浓度,乳酸脱氢酶(LDH)法检测各组(除锌原卟啉组外)细胞损伤程度,细胞划痕法检测正常对照组、碘美普尔组、三七皂苷+碘美普尔组细胞愈合率,酶标仪检测各组(除锌原卟啉组外)细胞中氧化应激指标[超氧化物歧化酶(SOD)、丙二醛(MDA)、活性氧(ROS)]含量,ELISA法检测各组(除锌原卟啉组外)细胞中炎症因子[白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)]含量,流式细胞术检测正常对照组、碘美普尔组、三七皂苷+碘美普尔组细胞凋亡率,RT-qPCR法检测各组细胞中核因子红细胞2相关因子2(Nrf2)、血红素氧合酶-1(HO-1)mRNA表达情况,ELISA法检测各组细胞中HO-1蛋白含量,免疫荧光检测细胞中Nrf2蛋白表达情况。结果细胞活力筛选,25μmol/L三七皂苷R1预处理24 h作为最优三七皂苷R1用药条件。与正常对照组比较,碘美普尔组细胞愈合率、细胞中SOD含量、细胞中HO-1 mRNA表达量及HO-1蛋白含量、Nrf2 mRNA表达量及Nrf2免疫荧光强度均明显降低(P均<0.05),LDH含量、MDA和ROS含量、IL-6和TNF-α含量、细胞凋亡率均明显升高(P均<0.05)。与碘美普尔组比较,三七皂苷+碘美普尔组的细胞活力、细胞愈合率、细胞中SOD含量、细胞中HO-1 mRNA表达量及HO-1蛋白含量、Nrf2 mRNA表达量及Nrf2免疫荧光强度均明显升高(P均<0.05),LDH含量、MDA和ROS含量、IL-6和TNF-α含量、细胞凋亡率均明显降低(P均<0.05);与三七皂苷+碘美普尔组比较,锌原卟啉组中Nrf2Objective It is to study the effect of panax notoginseng saponin R1(NGR1)on the injury of rat renal tubular epithelial cells(NRK-52E)induced by Iomeprol 400 and to explore its regulatory mechanism.Methods NRK-52E cells were divided into 5 groups:normal control group cultured normally,NGR1 group cultured with NGR1,Iomeprol group cultured with Iomeprol 400,NGR1+Iomeprol group cultured with NGR1 firstly,then cultured with Iomeprol 400,zinc protoporphyrin group cultured with zinc protoporphyrin+NGR1 firstly,then cultured with Iomeprol 400.The cell viability was evaluated by CCK8 to screen the optimal protective concentration of NGR1,and the degree of cell damage in each group(except zinc protoporphyrin group)was detected by LDH method,the cell healing rates in the normal control group,Iomeprol group,NGR1+Iomeprol group were detected by cell scratch method,the contents of oxidative stress indexes[superoxide dismutase(SOD),malondialdehyde(MDA),reactive oxygen species(ROS)]in each group(except zinc protoporphyrin group)were detected by microplate plate,the levels of inflammatory factors[interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)]in each group(except zinc protoporphyrin group)were detected by ELISA,and the apoptosis rates in the normal control group,Iomeprol group,NGR1+Iomeprol group were detected by flow cytometry,the expression of nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)mRNA in the cells of each group was detected by RT-qPCR,the content of HO-1 protein in the cells of each group was detected by ELISA,and the expression of Nrf2 protein was detected by immunofluorescence.Results In cell viability screening,25μmmol/L NGR1 pretreatment for 24h as the optimal dosing condition.Compared with the normal control group,the cell viability,cell healing rate,the content of SOD in cells,the expression of HO-1 mRNA and HO-1 protein,Nrf2 mRNA expression in cells and Nrf2 immunofluorescence intensity of the Iomeprol group were all significantly decreased(all P<0.05),and the contents of LDH,

关 键 词：三七皂苷R1 NRK-52E细胞 碘美普尔400 核因子红细胞-相关因子2 血红素氧合酶-1 

分 类 号：R-332[医药卫生]