N-乙酰半胱氨酸对抗丙酮醛引起的心肌细胞损伤  

N-acetylcysteine protects H9c2 cells against injuries induced by methylglyoxal

作　　者：董小变[1] 吴娟[1,2] 庄晓东[1] 黄泽娜[1] 胡洵[1] 廖新学[1] 

机构地区：[1]中山大学附属第一医院心血管内科,广东广州510080 [2]江门市中心医院心血管内科,广东江门529030

出　　处：《中国病理生理杂志》2016年第3期398-404,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81270296);广东省财厅社会发展项目(No.2014SC107)

摘　　要：目的：观察N-乙酰半胱氨酸（N-acetylcysteine,NAC）对抗丙酮醛诱导的H9c2心肌细胞损伤及相关机制。方法：实验分为正常对照组、丙酮醛损伤组（不同浓度丙酮醛处理）、NAC＋丙酮醛组（NAC与丙酮醛共处理）、SP600125预处理＋丙酮醛组、NAC组和SP600125组。H9c2心肌细胞常规消化种板,经相应处理24 h后：应用CCK-8法检测心肌细胞的存活率;Western blot法检测H9c2心肌细胞内磷酸化和总的c-Jun氨基端激酶（p-JNK、t-JNK）表达水平;双氯荧光素（DCFH-DA）染色法检测心肌细胞内活性氧（ROS）水平;罗丹明123（Rh123）染色法检测细胞线粒体膜电位（MMP）;Hoechst 33258染色法观察H9c2心肌细胞凋亡形态学变化。结果：与对照组相比,不同浓度的丙酮醛均能够降低H9c2心肌细胞存活率,且呈剂量依赖性（P〈0.01）,NAC在一定浓度范围内（500-1 500μmol/L）可对抗丙酮醛引起心肌细胞损伤（P〈0.01）,抑制丙酮醛引起细胞内ROS水平升高,对抗丙酮醛引起细胞内MMP降低,抑制丙酮醛诱导细胞内JNK蛋白的磷酸化（P〈0.01）。与NAC的细胞保护作用类似,选择性JNK抑制剂SP600125也可抑制丙酮醛诱导的细胞损伤,包括减轻氧化应激、改善线粒体膜电位及抑制细胞凋亡。结论：N-乙酰半胱氨酸能够保护H9c2心肌细胞对抗丙酮醛引起的损伤,其机制可能与其降低细胞内ROS水平、改善MMP、抑制JNK磷酸化和抗凋亡有关。AIM：To investigate the protective effect of N-acetylcysteine（ NAC） on H9c2 cells from injuries induced by methylglyoxal（ MG） and the potential mechanism.METHODS：H9c2 cells were divided into control group,MG treatment group,NAC ＋ MG treatment group,SP600125 pretreatment ＋ MG group,NAC group and SP600125 group.The viability of the H9c2 cells was measured by CCK-8 assay.The protein levels of p-JNK and t-JNK were tested by Western blot.The changes of intracellular reactive oxygen species（ ROS） were evaluated by 2＇,7＇-dichlorofluorescein diacetate（ DCFH-DA） staining.Mitochondrial membrane potential（ MMP） was measured by rhodamine 123（ Rh123） staining.The morphological changes in apoptotic cardiomyocytes were detected by Hoechst 33258 staining.RESULTS：During 100 - 800 μmol/L concentration range,MG caused significantly reduced viability of the H9c2 cells in a dose-dependent manner.NAC had a protective effect on H9c2 cells against the injuries induced by MG during 500 - 1 500 μmol/L concentration range through raising cell viability,inhibiting cellular oxidative stress and improving MMP（ P 〈0.01）.SP600125,an inhibitor of JNK,showed the protective effect similar to NAC on H9c2 cells against MG-induced injuries,including attenuating oxidative stress,improving MMP and suppressing apoptosis.CONCLUSION：N-acetylcysteine offers obvious protective effect on H9c2 cells against the injuries induced by methylglyoxal.The underlying mechanisms may be associated with decreasing the production of ROS,ameliorating MMP,inhibiting the activation of JNK and suppressing apoptosis.

关 键 词：H9C2细胞 N-乙酰半胱氨酸 丙酮醛 C-JUN氨基端激酶 

分 类 号：R363[医药卫生—病理学]