摘要
目的观察蛋白质非酶糖化是否会增加心肌缺氧/复氧损伤的敏感性。方法常规培养H9C2细胞,分别给予PBS(对照组)、丙酮醛(200μmol/L)、丙酮醛(200μmol/L)+氨基胍(100μmol/L)孵育6 d后接受缺氧/复氧处理,以MTT法测定心肌细胞存活率,微量酶活性测试法检测乳酸脱氢酶(LDH)释放变化,比色法检测丙二醛(MDA)含量,并通过免疫组织化学检测心肌细胞终末糖化晚期产物(AGEs)表达。结果缺氧/复氧降低细胞存活率,增加心肌细胞MDA含量和LDH释放;而丙酮醛处理6 d的心肌细胞的AGEs含量明显增加,并对缺氧/复氧损伤的敏感性增加,与PBS孵育细胞经缺氧/复氧处理后相比,MDA含量和LDH释放显著上升,存活率明显下降(P<0.01);MGO引起的上述变化可以被糖化抑制剂氨基胍部分阻断(P<0.05)。结论蛋白质非酶糖化是糖代谢中间产物丙酮醛增加心肌细胞对缺氧/复氧损伤敏感性的重要机制。
Objective To investigate whether protein glycation increases the injury of myocardial cells subjected to hypoxia/reoxygenation. Methods The cultured H9C2 cells were incubated for 6 days with PBS, Methylglyoxal, and Methylglyoxal＋Aminoguanidine, respectively. Then the cells were treated by hypoxia/reoxygenation, and the survival rates of cardiomyocytes in each group were detected by MTT method. LDH release and MDA content were determined in each group. AGEs contents in cells treated with PBS, MGO or MGO＋AG were assayed by immunohistochemical staining. Results Preculturing cardiomyocytes with MG for 6 days caused a significant increase in AGEs production, which was dramatically reduced by cotreatment with AG, a strong AGEs formation inhibitor, and preculturing cardiomyocytes with MG for 6 days made cardiomyocytes more susceptible to H/R, as evidenced by decreased survival rate of cells and LDH release. Conclusion Protein glycation sensitized cultured cardiomyocytes to H/R injury, and therapeutic interventions inhibitting AGEs formation may attenuate ischemic-reperfusion injury in hyperglycemic state diseases such as diabetes.
出处
《中华临床医师杂志（电子版）》
CAS
2013年第24期207-209,共3页
Chinese Journal of Clinicians(Electronic Edition)
基金
国家自然科学基金(81170144
81100150)
关键词
肌细胞
心脏
非酶糖化
缺氧
复氧
Myocytes, cardiac
Glyeation
Hypoxia/reoxygenation
