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目的:研究芒果甙的抗氧化性对柔红霉素心肌细胞毒性的保护作用及可能机制。方法:以4μmol/L柔红霉素(DNR)制作心肌细胞损伤模型,加入不同浓度芒果甙作用后,用MTT法检测心肌细胞存活率(Viability);用全自动生化仪检测培养基中乳酸脱氢酶(LDH)、肌酸激酶(CK)、肌酸激酶同工酶(CK-MB);用分光光度计检测超氧化物歧化酶(SOD)及丙二醛(MDA)含量;用流式细胞术(FCM)检测细胞内活性氧(ROS)及线粒体膜电位(Δm)水平;用荧光定量PCR检测热休克蛋白70(Hsp70)及过氧化氢酶(CAT)基因表达变化。结果:浓度为4μmol/L柔红霉素能明显损伤心肌细胞,MTT法及心肌酶检测显示芒果甙作用浓度为25μmol/L时即有保护作用。100μmol/L芒果甙组与柔红霉素模型组、维生素C对照组比较,心肌细胞存活率明显提高;心肌酶LDH、CK、CK-MB漏出减少,SOD消耗和MDA生成减少;心肌细胞内ROS浓度明显减少;100μmol/L芒果甙与柔红霉素模型组、维生素C对照组比较能明显恢复Δm水平,而维生素C组与柔红霉素组比较无显著差异。芒果甙随浓度增加,HSP70、CAT基因的表达水平逐渐降低。结论:在体外试验中,芒果甙能减轻柔红霉素所致的心肌细胞毒性,其机制可能与芒果甙有效清除氧自由基,减轻过氧化损伤有关。
Objective: To study the protective effect of mangiferin on cardiomyocyte cytotoxicity induced by daunorubicin and its possible mechanism. Methods: Cardiomyocytes were induced by 4μmol / L daunorubicin (DNR). After adding different concentrations of mangiferin, the viability of cardiomyocytes was measured by MTT assay. The lactate LDH, CK and CK-MB. The contents of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by spectrophotometer. The level of reactive oxygen species (ROS) and mitochondrial membrane potential (Δm) were measured by FCM. The changes of Hsp70 and CAT gene expression were detected by real-time fluorescence quantitative PCR. Results: Daunorubicin at a concentration of 4μmol / L could obviously damage myocardial cells. MTT assay and myocardial enzyme test showed that the protective effect of mangiferin was 25μmol / L. The survival rate of myocardial cells was significantly increased in 100μmol / L mangiferin group compared with daunorubicin model group and vitamin C control group. The leakage of myocardial enzymes LDH, CK and CK-MB was decreased, while the SOD consumption and MDA production were decreased. The concentration of 100μmol / L mangiferin and daunorubicin model group, vitamin C control group can significantly restore the level of Δm, while the vitamin C group and daunorubicin group no significant difference. With the increase of the concentration of mangiferin, the expression levels of HSP70 and CAT genes decreased gradually. Conclusion: In vitro, mangiferin can attenuate daunorubicin-induced cardiomyocyte cytotoxicity. Its mechanism may be related to the effective clearance of oxygen free radicals by mangiferin and the reduction of peroxidation injury.