人参皂苷Rg1对原代培养心肌细胞缺氧复氧损伤的作用

Protection effect of ginsenoside Rg1 on neonatal rat cardiomyocyte from hypoxia/reoxygenation injury

目的探讨人参皂苷Rg1对原代培养乳鼠心肌细胞缺氧复氧损伤的作用及其是否能直接抑制线粒体通透性转换孔的开放。方法原代培养大鼠乳鼠心肌细胞,构建缺氧复氧损伤(H/R)模型,应用不同浓度人参皂苷Rg1进行干预,通过检测细胞活力、线粒体膜电位、细胞色素C的释放,观察人参皂苷Rg1对心肌细胞H/R损伤的作用;将不同浓度的人参皂苷Rg1分别和线粒体25℃孵育10 min,加入200μmol/L CaCl_2诱发mPTP的开放,以1μmol/L的mPTP特异性抑制剂环孢素A(Cs A)为阳性对照,每间隔30 s观察520 nm处吸光度的变化,连续观察10 min,以线粒体肿胀程度评估mPTP的开放情况。结果人参皂苷Rg1能增强H/R损伤后心肌细胞活力,阻止H/R引起的线粒体膜电位的降低,减少细胞色素C释放;6.25,25,100,400μmol/L人参皂苷Rg1无直接抑制mPTP开放的作用。结论人参皂苷Rg1可减轻心肌细胞H/R损伤,保护心肌细胞,其作用与直接抑制mPTP开放无关。

Objective It is to investigate the effect of ginsenoside Rg1 on hypoxia-reoxygenation damage of primary cultured neonatal rat myocardial cell and that whether ginsenoside Rg1 could directly inhibit the opening of mitochondrial permeability transition pore. Methods Hypoxia-reoxygenation( H/R) injury model on primary cultured neonatal rat myocardial cells were established. Then,the effect of ginsenoside Rg1 was determined by examining cell vitality,mitochondrial transmerabrane potential and the release of cytochrome C. Different concentrations of ginsenoside Rg1 were respectively incubated with mitochondria at 25 ℃ for 10 min,followed by the addition of 200 μmol/L CaCl_2 to induce the opening of mPTP,with 1 μmol/L cyclosporin A,a specific inhibitor of mPTP as a positive control. The change of absorbance on 520 nm was continuously observed every 30 s,for 10 min,which reflected the mitochondria swelling degree and indicated the opening of mPTP. Results Ginsenoside Rg1 could enhance the vitality of myocardial cells after H/R injury,prevent the reduction of mitochondrial transmerabrane potential caused by H/R injure,decrease the release of cytochrome C,and protect the myocardial cell. 6. 25,25,100,400 μmol/L Ginsenoside Rg1 could not directly restrain the opening of mPTP. Conclusion Ginsenoside Rg1 relieved the injury of myocardial cells caused by H/R and protected the myocardial cell,which had nothing to do with the direct inhibition of mPTP opening.