N-乙酰半胱氨酸对丙酮醛诱导的人肾小管上皮细胞损伤的保护作用及机制研究

摘要

Objective To investigate the protective effect of n-acetylcysteine (NAC) on injury of human renal tubular epithelial cells (HK-2 cells) induced by methylglyoxal and its underlying mechanism. Methods HK-2 cells were divided into six group: control group (no treatment), injury group (induced by methylglyoxal) and NAC groups (treated with different concentrations of NAC, 1, 2, 4, 8 mmol/L). Cell viability was measured by CCK-8. Morphological changes of apoptotic cell nucleus were observed by DAPI staining. Mitochondrial membrane potential (MMP) was detected by rhodamine 123 staining and flow cytometry. Expression of Bax, Bcl-2, Caspase-3 and p-ERK1/2 were identified by western blot. Results Compared with control group, cell viability, MMP and the expression of Bcl-2 of HK-2 cells were significantly reduced by methylglyoxal while the number of apoptotic cells and the protein level of Bax, Cleaved Caspase-3 and p-ERK1/2 were dramatically increased. However, all of the above changes were reversed by NAC intervention. Conclusions NAC exerts protective effect on methylglyoxal induced injury of HK-2 cells. Potential mechanism may be associated with the anti-apoptotic effect and inhibition of activation of ERK1/2 cell signaling.%目的 探讨N-乙酰半胱氨酸(NAC)对丙酮醛引起人肾小管上皮细胞(HK-2细胞)损伤的保护作用及其可能机制.方法 HK-2细胞分为6组:对照组、丙酮醛组、丙酮醛+不同浓度NAC(1、2、4及8 mmol/L)处理组.采用CCK-8检测细胞活力,DAPI染色观察细胞核形态,罗丹明123染色及流式细胞仪检测细胞线粒体膜电位,Western blot检测抗凋亡蛋白Bcl-2、促凋亡蛋白Bax、Cleaved Caspase-3以及磷酸化ERK1/2(p-ERK1/2)蛋白表达水平.结果 与对照组比较,HK-2细胞经400 μmol/L丙酮醛处理后,细胞活力、线粒体膜电位及Bcl-2蛋白表达水平降低,凋亡细胞数增多,Bax、Cleaved Caspase-3及p-ERK1/2蛋白表达水平则升高.而经不同浓度的NAC干预后,可以逆转丙酮醛对HK-2细胞的上述作用.结论 NAC对丙酮醛诱导的HK-2细胞损伤具有保护作用,其机制可能与其抗凋亡及抑制ERK1/2信号通路有关.