摘要
目的探讨黄芪甲苷(AS-Ⅳ)对高糖(HG)环境下人肾小球系膜细胞(HMCs)增殖和氧化应激损伤保护作用及其可能机制。方法采用MTT方法检测高糖作用不同时间点时及不同浓度AS-Ⅳ干预后HMCs增殖情况;采用实时荧光定量PCR(qPCR)、Western blot方法分别检测不同浓度AS-Ⅳ干预高糖环境下HMCs 48 h后核因子E2相关因子2(Nrf2)、血红素氧合酶1(HO-1)、诱导型一氧化氮合酶(iN-OS)、细胞间黏附分子-1(ICAM-1)mRNA及其蛋白表达;采用过氧化氢(H_2O_2)、总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)试剂盒分别检测不同浓度AS-Ⅳ干预高糖环境下HMCs 48 h后细胞上清液中H_2O_2、T-SOD、GSH-Px、MDA含量变化。结果 MTT法结果显示,与正常(NG)组比较,高糖环境下HMCs呈过度增殖趋势(P<0.05),不同浓度AS-Ⅳ干预48 h后,与HG组比较,各用药组明显地抑制了高糖环境下HMCs过度增殖且呈剂量依赖性(P<0.01);qPCR及Western blot法结果显示,与HG组比较,各用药组明显地增加了Nrf2、HO-1 mRNA及蛋白表达(P<0.01),减少了iN OS与ICAM-1 mRNA及蛋白表达(P<0.05)。各试剂盒测试结果显示,与HG组比较,各用药组明显地降低了HMCs上清液中H_2O_2、MDA的含量(P<0.05),增加了HMCs上清液中T-SOD、GSH-Px的含量(P<0.01)。结论 AS-Ⅳ能够明显地抑制HG环境下HMCs过度增殖。对HG环境下HMCs损伤保护作用的部分机制可能是通过激活Nrf2通路并上调Nrf2、HO-1 mRNA及其蛋白其表达,下调iN OS、ICAM-1 mRNA及其蛋白表达,增加HMCs上清液中T-SOD、GSH-Px的含量,降低HMCs上清液中H_2O_2、MDA的含量。
Objective To study the effects of astragaloside IV in high glucose-induced proliferation and oxidative stress in HMCs and its mechanism. Methods HMCs were cultured in high glucose with or without different concen- tration of AS-IV. After 48 h, the effect of AS-IV in HMCs proliferation were detected by MTT assay. The mRNA ex- pression and protein levels of Nrf2, HO-1, iNOS, ICAM-1 were determined by qPCR and Western blot, respective- ly. The supernatant were collected to measure the amount of H202,MDA and the activity of T-SOD and GSH-Px. Results Compared with normal group, MTT showed that HMCs significantly increased in high glucose conditions （P 〈0.05）. Compared with high glucose group （HG）, AS-IV significantly inhibited the proliferation of HMCs in dose-dependent manner（P 〈 0. 01 ）. Western blot and qPCR showed that AS-IV increased Nrf2 and HO-1 expres- sion（P 〈 0. 01 ） and suppressed the overexpression of iNOS and ICAM-1 （P 〈 0.05 ） in HMCs in high glucose con- ditions, reduced the amount of H2 02 and MDA（P 〈 0.05）, and increased the activity of T-SOD and GSH-Px（P 〈 0.01 ） in supernatant. Conclusion AS-ivcould significantly inhibit HMCs excessive proliferation in high glucose conditions. The mechanism maybe related to the activating of Nrf2 pathway, upregulating Nrf2, HO-1 protein and mRNA expression, downregulating protein and mRNA expression of iNOS, ICAM-1, increasing in cell supernatant on T-SOD and GSH-Px,reducing H202 and MDA content in the supernatant.
出处
《安徽医科大学学报》
CAS
北大核心
2017年第7期972-978,共7页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81173624)
安徽省国际科技合作计划项目(编号:12030603007)
安徽省高校省级自然科学研究重大项目(编号:KJ2016SD35)
