姜黄素联合瑞舒伐他汀对大鼠心肌梗死损伤的保护作用机制的研究

Study on the protective mechanism of curcumin combined with rosuvastatin on myocardial infarction

目的研究姜黄素联合瑞舒伐他汀对大鼠心肌梗死损伤的保护作用以及可能作用机制。方法用结扎左冠脉前降支的方法构建心肌梗死大鼠模型。40只大鼠随机分为假手术组、心肌梗死组、姜黄素组、瑞舒伐组、姜黄素+瑞舒伐组。假手术组实行开胸术,但不结扎左冠脉前降支。冠脉结扎24 h后,以灌胃的方式分别给予大鼠姜黄素100mg/(kg·d)、瑞舒伐他汀10 mg/(kg·d)、姜黄素100 mg/(kg·d)+瑞舒伐他汀10 mg/(kg·d),假手术组、心肌梗死组以同样的方式给予等量生理盐水,连续给药4周。实验结束后以超声仪检测左心室大小如左室舒张末期内径(LVIDd)、左室收缩末期内径(LVIDs)、左室射血分数(EF);大鼠处死后,测量大鼠体重、心脏重量以及比值;采用末端脱氧核昔酸转移酶(Td T)介导的原位末端标记法(TUNEL)检测心肌梗死组织石蜡切片心肌细胞凋亡指数;蛋白质印迹法(Western Blot)检测心肌组织中凋亡相关蛋白Fas、含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)的表达量。结果与假手术组相比,心肌梗死组大鼠LVIDd、LVIDs、心脏重量、心脏/体重显著增加(P<0.05),EF、体重显著降低(P<0.05),表明大鼠心脏收缩、舒张功能障碍,心功能损伤,造模成功;瑞舒伐他汀和姜黄素显著降低LVIDd、LVIDs、心脏重量、心脏/体重(P<0.05),EF、体重显著增加(P<0.05),表明瑞舒伐他汀和姜黄素可改善心肌梗死损伤,联合作用效果更显著;大鼠发生心肌梗死后心肌细胞的凋亡指数显著上调(P<0.05),Fas、Caspase-3蛋白表达量显著上调(P<0.05),姜黄素和瑞舒伐他汀可缓解心肌细胞的凋亡以及Fas、Caspase-3蛋白增加,联合作用效果优于单药的作用。结论姜黄素和瑞舒伐他汀可缓解大鼠心肌梗死损伤,改善心功能,可能通过抑制Fas、Caspase-3蛋白水平进而阻碍心肌细胞凋亡发挥作用。

Objective To investigate the protective effect of curcumin combined with rosuvastatin on myocardial infarction and its possible mechanism. Methods The 40 rats models of myocardial infarction were established by ligating the left coronary artery. Rats were randomly divided into sham operation group,myocardial infarction group,curcumin group,rosuvastatin group,curcumin ＋ rosuvastatin group; Sham operation group underwent thoracotomy,but not ligation of left coronary artery. After coronary ligation for 24 h,the rats were given 100 mg/（ kg·d） curcumin,10 mg/（ kg·d） rosuvastatin,100 mg/（ kg·d） curcumin ＋ 10 mg/（ kg·d） rosuvastatin,respectively. The sham operation group and the myocardial infarction group were given the same amount of normal saline in the same way for 4 weeks. At the end of the experiment,Left ventricle internal diameter at end-diastolic phase（ LVIDd）,LV internal diameter at end-systolic phase（ LVIDs） and ejection fraction（ EF） were measured by ultrasound. After sacrificed,the weight,heart weight and the ratio were measured. The apoptotic indexes of paraffin sections of myocardial infarction were detected by Transferase-mediated d UTP nick end labeling（ TUNEL）. Western blot was used to detect the expression of Fas and caspase-3 in myocardial tissue. Results Compared with the sham operation group,LVIDd,LVIDs,heart weight,heart/body weight of rats in myocardial infarction group were significantly increased（ P〈0. 05）,and EF,body weight were significantly decreased（ P〈0. 05）,indicating that cardiac systolic and diastolic dysfunction,and cardiac function damaged,the rat model of myocardial infarction were successful. Rosuvastatin and curcumin significantly decreased LVIDd,LVIDs,heart weight,heart/body weight（ P〈0. 05）,and EF and body weight were significantly increased（ P〈0. 05）,indicating that rosuvastatin and curcumin could improved myocardial infarction,and the combined effect was more significant. The apoptotic index of cardiomyocytes,Fas and Caspase-3 protein level were significantly up-regulated（ P〈0. 05） after myocardial infarction in rats（ P〈0. 05）. Curcumin and rosuvastatin coula relieved the apoptotic of cardiomyocytes and Fas and Caspase-3 protein level,and the combined effect due to single drug effect. Conclusion Curcumin and rosuvastatin can alleviate myocardial infarction injury and improve cardiac function in rats,which may by inhibiting Fas and Caspase-3 protein level and thus hinder the myocardial cell apoptosis.