芒果甙对柔红霉素致大鼠心肌细胞过氧化损伤的保护作用

Protective effect of mangiferin on daunorubicin-induced rat cardiomyocytes peroxidation

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 daunorubicin-induced rat cardiomyocytes peroxidation and the possible mechanism.Medthods：Creat cardiomyocytes injury model through 4μmol/L DNR,then treat with different concentrations of Mangiferin to detect cardiomyocytes viability by MTT assay;to detect LDH,CKand CK-MB by Automatic biochemical analyzer;to detect the content of SOD and MDA by spectrophotometer,to detect the level ROS andΔ m by FCM;determine Hsp70 and Catalase CAT gene expression change by fluorescence quantitative PCR.Results：The 4μmol/L DNR model group can injure cardiomyocytes obviously and mangiferin can protect cardiomyocytes as long as it＇s concentration reachs 25μmol/L.Compaired with DNR model group and VitC group,100μmol/L mangiferin group can improve cardiomyocyte viability obviously（P0.01）,reduce LDH、CK and CK-MB leakage（p0.01）,reduce SOD consumption and MDA production and reduce ROS in cardiomyocyt（P0.01）.Compaired with DNR model group and VitC group,100μmol/L mangiferin group can restore Δ m effectively（P0.01）.However,the effect difference on restoring Δ m between DNR model group and VitC group has no stastical significance（P=0.368）.With the concentration of mangiferin increasing,the expression of HSP70 and CAT gene decreasing.Conclusion： In vitro,Mangiferin can relieve myocardial toxicity induced by daunorubicin,the possible mechanism is related with mangiferin can effectively clear the ROS and relieve peroxidation.