强心一号复方对脓毒症小鼠心肌细胞凋亡的影响

Effect of Qiangxin1 Decoction on Apoptosis of Myocardial Cells in Sepsis Mice

目的从心肌细胞凋亡及凋亡相关蛋白表达调节的角度探讨强心一号复方对脓毒症小鼠的心脏保护机制。方法取36只健康雄性C57BL/6小鼠,随机分为假手术组、模型组、强心组,每组12只。模型组和强心组采取盲肠结扎穿孔术(CLP)制备脓毒症模型。强心组于术后2 h给予1 g/kg强心一号药液灌胃,模型组和假手术组给予等量0.9%氯化钠注射液灌胃。采用临床分级评价术后临床指数,麦胚凝集素(WGA)染色评价心肌细胞肥大,原位末端标记法(TUNEL)评价心肌细胞凋亡,蛋白免疫印迹法检测凋亡相关蛋白B细胞淋巴瘤因子2(Bcl-2)、Bcl-2相关X蛋白(Bax)和裂解的半胱氨酸天冬氨酸蛋白酶3(Cleaved Caspase3)的蛋白表达水平。结果与假手术组相比,模型组临床评分显著下降(P<0.05),心肌细胞代偿性肥大(P<0.05),心脏组织TUNEL阳性细胞增多(P<0.05),Cleaved Caspase3蛋白表达显著增高(P<0.05)。与模型组相比,强心组临床评分显著改善(P<0.05),心肌细胞代偿性肥大情况改善(P<0.05),抗凋亡蛋白Bcl-2蛋白表达显著增高(P<0.05),促凋亡蛋白Bax和Cleaved Caspase3蛋白表达显著下降(P<0.05)。结论强心一号复方可抑制CLP小鼠心肌细胞凋亡从而发挥心脏保护作用,其机制可能与调节凋亡相关蛋白Bcl-2、Bax和Cleaved Caspase3的表达有关。

Objective:To explore the cardioprotective mechanism of Qiangxin1 Decoction on sepsis mice from the perspective of apoptosis and expression regulation of apoptosis-related proteins.Methods:36 male C57 BL/6 mice were randomly divided into sham-operated group,the model group and Qiangxin group,12 mice in each.Cecal ligation and puncture(CLP)was used to establish sepsis model in the model group and Qiangxin group.Qiangxin group was given 1 g/kg Qiangxin1 Decoction 2 hours after operation,while the sham-operated was given0.9%sodium chloride injection.Postoperative clinical index was assessed by clinical grading.Myocardial hypertrophy was assessed by WGA staining.Cardiomyocyte apoptosis was assessed by TUNEL.The protein expression of Bcl-2,Bax and Cleaved Caspase 3 were detected by Western blotting.Results:Compared with the sham operation group,the clinical score of the model group decreased significantly(P<0.05).Cardiomyocytes was compensatory hypertrophy(P<0.05).TUNEL positive cells and the protein expression of Cleaved Caspase 3 were significantly increased(P<0.05).Compared with the model group,the clinical score and compensatory hypertrophy of cardiac myocytes were significantly improved in Qiangxin group(P<0.05).The protein expression of anti-apoptotic protein Bcl-2 was significantly increased(P<0.05),and the expression of pro-apoptotic protein Bax and Cleaved Caspase 3 was significantly decreased(P<0.05).Conclusion:Qiangxin1 Decoction can inhibit the apoptosis of cardiac myocytes and play a protective role in the heart of CLP mice.The mechanism may be related to the regulation of the expression of apoptosis-related proteins Bcl-2,Bax and Cleaved Caspase3.