搜风祛痰中药对心肌缺血再灌注大鼠冠脉微循环血管内皮损伤及凋亡相关蛋白的影响

Effect of Chinese herbal medicine for searching wind and expelling phlegm on vascular endothelium and apoptosis-related protein expression for coronary microcirculation in rats with myocardial ischemia-reperfusion injury

目的探讨搜风祛痰中药对心肌缺血再灌注大鼠冠脉微循环血管内皮损伤及凋亡相关蛋白的影响。方法将140只8周龄SPF级雄性SD大鼠随机分成假手术组、模型组、培哚普利组、搜风祛痰中药组,每组35只。假手术组、模型组大鼠予以生理盐水10 mL/kg灌胃,培哚普利组大鼠予以培哚普利片0.4 mg/kg灌胃,搜风祛痰中药组大鼠予以搜风祛痰中药8.1 g/kg灌胃,均每日1次,连续灌胃7 d。末次灌胃结束后1 h建立心肌缺血再灌注模型,假手术组只穿线不结扎,心电图评价造模情况。造模成功后ELISA法检测各组大鼠血清可溶性内皮细胞蛋白C受体(sEPCR)、可溶性血栓调节蛋白(sTM)水平,Western Blot法检测心肌组织中蛋白激酶B(Akt)、磷酸化蛋白激酶B(P-Akt)、B细胞淋巴瘤/白血病2关联X(Bax)、B细胞淋巴瘤/白血病2(Bcl-2)蛋白表达水平。结果模型组大鼠血清sTM、sEPCR水平和心肌组织中Bax、Bcl-2蛋白表达水平均明显高于假手术组(P均<0.05),心肌组织中Akt、P-Akt蛋白表达水平及Bcl-2/Bax比值均明显低于假手术组(P均<0.05);培哚普利组和搜风祛痰中药组大鼠血清sTM、sEPCR水平和心肌组织中Bax蛋白表达水平均明显低于模型组(P均<0.05),心肌组织中P-Akt、Bcl-2蛋白表达水平和Bcl-2/Bax比值明显高于模型组(P均<0.05);培哚普利组大鼠心肌组织中Akt蛋白表达水平明显高于模型组(P<0.05),但搜风祛痰中药组Akt蛋白表达水平与模型组比较差异无统计学意义(P>0.05)。结论搜风祛痰中药具有抑制凋亡、保护心肌缺血再灌注大鼠冠脉微循环血管内皮损伤作用,其机制可能与降低sTM、sEPCR水平,上调P-Akt、Bcl-2蛋白和下调Bax蛋白表达相关。

Objective It is to investigate the effect of Chinese herbal for searching wind and expelling phlegm on vascular endothelium and apoptosis-related protein expression for coronary microcirculation in rats with myocardial ischemia-reperfusion injury.Methods A total of 8 weeks old 140 female SPF Sprague-Dawley rats for were randomly divided into sham operation group,model group,perindopril group,searching wind and expelling phlegm Chinese herbal group(Chinese medicine group),35 rats in each group.The rats in the sham operation group and model group were given normal saline 10 mL/kg by intragastric administration,the rats in the perindopril group were given perindopril tablets 0.4 mg/kg by intragastric administration,and the rats in the Chinese medicine group were given Chinese herbal for searching wind and expelling phlegm 8.1 g/kg by intragastric administration,all once per day,continuously treated for 7 days.A model of myocardial ischemia-reperfusion was established in 1 h after the last gastric gavage.The sham-operated group was only threaded without ligation,and the electrocardiogram was used to evaluate the modeling.After successful modeling,ELISA method was used to detect the levels of soluble endothelial protein C receptor(sEPCR)and soluble thrombomodulin(sTM)in the rats of every group.Western Blot method was used to detect the expression levels of protein kinase B(Akt)and phosphorylated protein kinase B(P-Akt),B-cell lymphoma/leukemia 2 associated X(Bax),B-cell lymphoma/leukemia 2(Bcl-2)in myocardial tissue.Results The levels of serum sTM,sEPCR and the expression of Bax and Bcl-2 protein in the myocardial tissue in the model group were significantly higher than those in the sham operation group(all P<0.05),while the expression levels of Akt and P-Akt protein in the myocardial tissue and Bcl-2/Bax ratio were significantly lower than those in the sham operation group(all P<0.05).The levels of serum sTM,sEPCR and the expression of Bax protein in the myocardial tissue in the perindopril group and the Chinese medicine group were significantly lower than those in the model group(all P<0.05),the expression levels of P-Akt and Bcl-2 protein in the myocardial tissue and Bcl-2/Bax ratio were significantly higher than those in model group(all P<0.05);The protein expression level of Akt in myocardial tissue in perindopril group was significantly higher than that in the model group(P<0.05),but there was no statistically significant difference in the expression level of Akt protein between the Chinese medicine group and model group(P>0.05).Conclusion The Chinese herbal for searching wind and expelling phlegm can inhibite apoptosis and protect vascular endothelium of coronary microcirculation in rats with myocardial ischemia-reperfusion injury.The mechanism may be related to its decreasing of levels of sTM,sEPCR in serum,up-regulation of P-Akt,Bcl-2 proteins and down-regulation of Bax proteins.