血脂康对SD大鼠心脏缺血/再灌注心肌细胞凋亡的影响

Effects of Xuezhikang on Cardiomyocyte Apoptosis in Cardiac Ischemia/Reperfusion SD Rats

目的探讨血脂康预处理对SD大鼠在体心脏缺血/再灌注后心肌细胞凋亡的影响。方法40只SD大鼠随机分成假手术组、模型对照组、甲羟戊酸组、血脂康组及血脂康+甲羟戊酸组,各8只。假手术组:双蒸水,灌胃1周;模型对照组:双蒸水2 mL灌胃1周;甲羟戊酸组:甲羟戊酸150 mg·kg^(-1)·d^(-1)溶于2 mL双蒸水,灌胃1周;血脂康组:血脂康原粉2.8 g·kg^(-1)·d^(-1)溶于2 mL双蒸水,配成悬液灌胃1周:血脂康+甲羟戊酸组:血脂康原粉2.8 g·kg^(-1)·d^(-1)及甲羟戊酸150 mg·kg^(-1)·d^(-1)溶于2 mL双蒸水,灌胃1周。1周后结扎左冠状动脉前降支,结扎30 min后松开,恢复冠状动脉血流,复制在体心脏急性缺血/再灌注模型,假手术组仅穿线而不结扎前降支,其他手术步骤相同。再灌注120 min后,各组大鼠应用心脏原位末端转移标记技术(TUNEL)检测心肌细胞凋亡,实时荧光定量PCR和蛋白免疫印迹法检测脂肪酸合成酶(Fas)和Fas配体(FasL)mRNA和蛋白表达。结果假手术组中的SD大鼠心肌细胞偶有凋亡,缺血-再灌注损伤引起大鼠心肌细胞凋亡增加。血脂康组与模型对照组、血脂康+甲羟戊酸组、甲羟戊酸组比较,大鼠心肌细胞凋亡均减少,差异有统计学意义(P<0.01),但仍较假手术组大鼠心肌细胞凋亡增多,差异有统计学意义(P<0.01);而血脂康+甲羟戊酸组、甲羟戊酸组和模型对照组大鼠心肌细胞凋亡比较差异无统计学意义(P>0.05)。血脂康组与模型对照组比较,可减少Fas和FasL mRNA及蛋白表达,差异有统计学意义(P<0.01),甲羟戊酸可以减弱血脂康的保护效应(P<0.05)。结论血脂康抑制缺血/再灌注大鼠心肌细胞Fas/FasL凋亡通路,进而抑制心肌细胞凋亡。

Objective To investigate the effect of Xuezhikang on myocardial cell apoptosis in SD rats after cardiac ischemia/reperfusion in vivo.Methods Forty SD rats were randomly divided into sham operation group,model control group,mevalonate group,Xuezhikang group and Xuezhikang+mevalonate group,with 8 rats in each group.Sham operation group was given double distilled water by gavage with for 1 week,and model control group was given 2 mL of double distilled water by gavage for 1 week,and mevalonate group was given 150 mg·kg^(-1)·d^(-1) mevalonate dissolved in 2 mL of double-distilled water by gavage for 1 week,and Xuezhikang group was given 2.8 g·kg^(-1)·d^(-1) Xuezhikang dissolved in 2 mL of double-distilled water by gavage for 1 week,and Xuezhikang+mevalonate group was given 2.8 g·kg^(-1)·d^(-1) Xuezhikang original powder and 150 mg·kg^(-1)·d^(-1) mevalonate dissolved in 2 mL of double distilled water by gavage for 1 week.One week later,the left anterior descending coronary artery was ligated,and the ligation was released after 30 min to restore coronary blood flow to replicate the acute ischemia/reperfusion model of the heart in vivo.After reperfusion for 120 min,cardiomyocyte apoptosis was detected by terminal deoxynucleotidyl transferase-mediated nick end labeling(TUNEL)in rat heart,and mRNA and protein expressions of fatty acid synthase(Fas)and Fas ligand(FasL)were detected by real-time fluorescent quantitative PCR and western blotting.Results Cardiomyocyte apoptosis was occasionally seen in the sham-operated group,and ischemia-reperfusion injury induced increased cardiomyocyte apoptosis in rats.Compared with the control group,Xuezhikang+mevalonate group and mevalonate group,myocardial cell apoptosis in the Xuezhikang group was reduced,and the difference was statistically significant(P<0.01),but it was still higher than that in the sham operation group(P<0.01).There was no significant difference in myocardial cell apoptosis between Xuezhikang+mevalonate group,mevalonate group and model control group(P>0.05).Compared with the control group,the myocardial Fas and FasL mRNA and protein expressions of the rats in the Xuezhikang group were decreased,and the difference was statistically significant(P<0.01).Mevalonate could attenuate the protective effect of Xuezhikang(P<0.05).Conclusion Xuezhikang can inhibit the Fas/FasL apoptosis pathway of myocardial cells in ischemia/reperfusion rats,thereby inhibiting myocardial cell apoptosis.