化痰通络法对rt-PA溶栓后急性脑梗死大鼠神经细胞凋亡及Caspase-12表达的影响

Effect on apoptosis of neural cell and Caspase-12 expression in rat cortex accepted Huatan Tongluo formula with thrombolysis pherapy after acute cerebral infarction

[目的]观察化痰通络方对急性脑梗死大鼠经重组组织纤溶酶原激活剂(rt-PA)溶栓治疗后内质网应激(ERS)神经细胞凋亡途径中半胱氨酸天冬氨酸蛋白酶12(Caspase-12)蛋白和基因表达及神经元凋亡的影响。[方法]选择160只健康的SD大鼠,随机分为假手术组、模型组、rt-PA组和实验组共4组。采用自身栓子法制备大鼠大脑中动脉栓塞模型(MCAO),实验组与rt-PA组大鼠经尾静脉注入rt-PA(浓度:5.67 mg/kg)溶栓治疗,并联合化痰通络法中药灌胃(浓度:7.2 g/kg),每日2次。每组分别于为6 h、24 h、3 d和7 d 4个时相,观察梗死区域脑组织神经细胞Caspase-12蛋白和基因的表达,及神经元细胞凋亡的情况。[结果]与假手术组相比,模型组各时相Caspase-12蛋白与基因的表达量均明显增加(P<0.05)。与模型组相比,除7 d时rt-PA组Caspase-12蛋白表达减少,差异无统计学意义(P>0.05)外,rt-PA组和实验组在其他各时相Caspase-12蛋白与基因的表达均明显减少(P<0.05)。与rt-PA组相比,实验组Caspase-12基因在1 d、7 d时表达量明显减少(P<0.05),而Caspase-12蛋白表达差异无统计学意义(P>0.05),但有下降趋势。[结论]化痰通络法中药可降低急性脑梗死大鼠溶栓治疗后梗死区域Caspase-12蛋白和基因的表达,该法可能通过影响该环节,进而部分抑制内质网应激后期神经细胞的凋亡,从而防治急性脑梗死溶栓后缺血/再灌注的发生与发展。

[Objective] To observe the effect of Huatan Tongluo therapy on the expression of Caspase-12 protein,m RNA and apoptosis of nerve cells in the late of endoplasmic reticulum stress(ERS) in rat cortex accepted thrombolysis therapy with recombinant tissue-type plasminogen activator(rt-PA) after the acute cerebral infarction.[Methods] The 160 healthy SD rats which were randomly divided into sham group,model group,rt-PA group and experimental group.By using the MCAO rat model,the rats in rt-PA group and experimental group were injected rt-PA(concentration of 5.67 mg/kg),and those of experiment group were given Chinese herb of Huatan Tongluo therapy(concentration of 7.2 g/kg),2 times a day,and each group was observed at four time points: 6 h,1 d,3 d and 7 d.And then respectively observe the gene expression of Caspase-12 protein and m RNA,and the apoptosis in rats' brain neurons.[Results] Compared with sham group,the expression of Caspase-12 protein and m RNA increased significantly(P <0.05).Compared with model group,the expression of Caspase-12 protein and m RNA increased significantly(P<0.05) except 7 d which had no significant difference(P>0.05).Compared with rt-PA group,the expression levels of Caspase-12 m RNA of experiment group at 1 d and 7 d were significantly lower,and the protein had no significant difference with others(P>0.05),but with down trend.[Conclusion] Huatan Tongluo therapy can reduce the activation of Caspase-12 protein and m RNA in ERS,and inhibit the apoptosis of nerve cells,prevent reperfusion injury after acute cerebra accepted thrombolysis.