丹参酮ⅡA对人肾小球系膜细胞晚期糖化终产物受体表达及氧化应激水平的影响研究

Effect of Tanshinone ⅡA on RAGE Expression and Oxidative Stress Status in Human Mesangial Cells Induced by AGE

目的探讨丹参酮ⅡA(TanⅡA)对晚期糖化终末产物(AGE)诱导人肾小球系膜细胞(HMCs)的晚期糖化终末产物受体(RAGE)表达和氧化应激水平的影响。方法常规方法培养HMCs,分为:对照组;AGE组〔晚期糖化终末产物牛血清清蛋白(AGE-BSA)1.0、10.0、50.0、100.0μg/ml〕;AGE+TanⅡA组(AGE-BSA50.0μg/ml,TanⅡA 0、0.1、1.0、5.0、10.0μg/ml),以GAPDH为内参照,分别采用Western blotting和实时定量PCR法检测RAGE和mRNA表达水平,同时检测细胞培养上清液超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和丙二醛(MDA)水平。结果对照组与不同浓度AGE组HMCs RAGE和mRNA表达比较,差异均有统计学意义(F值分别为4.428和5.031,P<0.05);其中与对照组比较,10.0、50.0、100.0μg/ml AGE组HMCs RAGE和mRNA表达水平升高(P<0.05)。对照组与AGE+不同浓度TanⅡA组HMCs RAGE和mRNA表达比较有差异(F值为5.002和5.312,P<0.05);其中与AGE+0μg/ml TanⅡA组比较,对照组、AGE+0.1μg/ml TanⅡA组、AGE+1.0μg/ml TanⅡA组、AGE+5.0μg/ml TanⅡA组、AGE+10.0μg/ml TanⅡA组HMCs RAGE和mRNA表达降低(P<0.05)。对照组与不同浓度AGE组及AGE+TanⅡA组上清液SOD、GSH-Px和MDA水平比较有差异(P<0.05)。结论 TanⅡA能够明显降低AGE诱导HMCs RAGE和mRNA的表达水平,同时氧化应激水平也得到明显改善。

Objective To investigate the effect of Tanshinone ⅡA（ TanⅡA） on the expression of receptor for advanced glycated endproducts（ RAGE） and the oxidative stress status of human mesangial cells induced by AGE. Methods Human mesangial cells（ HMCs） were cultured with various concentration of AGE- BSA（ 1. 0 μg /ml,10. 0 μg /ml,50. 0 μg /ml and 100. 0 μg /ml）,and then were treated with Tan ⅡA（ 0. 1 μg /ml,1. 0 μg /ml,5. 0 μg /ml and 10. 0 μg /ml）. The cultured HMCs were divided into AGE group,AGE ＋ TanⅡ A group and control group. By using GAPDH as reference,the protein and mRNA expression of RAGE were evaluated by Western blotting analysis and real- time PCR respectively. The activity of superoxide dismutase（ SOD）,glutathione peroxidase（ GSH- Px） and malonaldehyde（ MDA） levels in the supernatant of HMCs were measured. Results The expressions of RAGE and mRNA in HMCs between control group and different concentration of AGE groups all showed statistically significant differences（ F = 4. 428 and 5. 031,P〈0. 05）. Compared with the control group,the expressions of RAGE and mRNA in HMCs of 10. 0 μg /ml,50. 0 μg /ml and 100. 0 μg /ml AGE groups were significantly higher（ P〈0. 05）. The expressions of RAGE and mRNA in HMCs between control group and AGE ＋different concentrations of TanⅡ A groups all showed statistically significant differences（ F = 5. 002 and 5. 312,P〈0. 05）. Compared with AGE ＋ 0 μg /ml TanⅡA group,the expressions of RAGE and mRNA in HMCs of the control group,AGE ＋0. 1 μg/ml TanⅡ A group,AGE ＋ 1. 0μg/ml TanⅡ A group,AGE ＋5. 0 μg/ml TanⅡ A group and AGE ＋ 10. 0 μg/ml TanⅡ A group were significantly lower（ P〈0. 05）. The SOD,GSH- Px and MDA levels in the supernantant of HMCs showed statistically significant differences between the control group,different concentrations of AGE group and AGE ＋ TanⅡ A group（ P〈0. 05）. Conclusion TanⅡ A could significantly reduce the RAGE and mRNA expression in HMCs induced by AGE,and the oxidative stress levels are also improved.