化痰通络方对急性脑梗死大鼠rt-PA溶栓后神经细胞凋亡途径中内质网应激相关基因GADD153/CHOP与JNK1表达的影响

目的:观察化痰通络方对急性脑梗死大鼠重组组织纤溶酶原激活剂(rt-PA)溶栓后内质网应激(endoplasmic reticulum stress,ERS)后期神经细胞凋亡途径中生长抑制DNA损伤基因153(C/EBP homologous protein,GADD153/CHOP)和c-Jun氨基末端激酶-1(c-Jun NH2-terminal kinases-1,JNK1)基因表达的影响。方法:将160只健康SD大鼠随机分为假手术组,模型组,rt-PA组,化痰通络方联合rt-PA组(简称中药组)。采用自身栓子法制备大鼠大脑中动脉栓塞模型(MCAO),rt-PA组与中药组分别给予尾静脉注入rt-PA(5.67 mg·kg-1)及联合化痰通络中药(7.2 g·kg-1)干预,2次/d。于6 h,1,3,7 d 4个时相分别采用RT-PCR检测各组大鼠大脑皮质梗死区组织中GADD153/CHOP与JNK1 mRNA的表达,采用TUNEL法检测各组大鼠大脑神经元细胞的凋亡。结果:与假手术组比较,模型组在4个时相中GADD153/CHOP与JNK1基因表达均明显增高(P0.05),同一时相内与假手术组比较,模型组神经元凋亡显著升高(P0.05);与模型组比较,rt-PA组和中药组在4个时相中均可降低JNK1 mRNA的表达量(P0.05),同时rt-PA组在1 d时可降低GADD153/CHOP mRNA的表达,而中药组可在6 h和1 d时同时降低其表达,差异显著(P0.05),rt-PA组在1 d和7 d时相神经元凋亡明显低于模型组(P0.05),中药组各个时相神经元凋亡均显著降低(P0.05),但rt-PA组与中药组之间无明显差异。结论:化痰通络方可通过降低内质网应激后期神经细胞凋亡途径中GADD153/CHOP与JNK1 mRNA的表达,部分抑制神经细胞的凋亡,从而防治急性脑梗死溶栓后缺血再灌注的发生与发展。

Effect of Huatan Tongluo Formulae on Expression of GADD153/CHOP and JNK1 Related with Endoplasmic Reticulum Stress in Apoptosis Pathway in Rat Cortex Accepted Thrombolysis Pherapy After Acute Cerebral Infarction

Objective: To observe the effect of Huatan Tongluo formulae on expression of C/EBP homologous protein(GADD153/CHOP) and C-Jun NH₂-terminal Rinases(JNK1) in apoptosis pathway of endoplasmic reticulum stress in rat reconstructive tissue plasminogen activator(rt-PA) after the acute cerebral infarction. Method: The 160 healthy male SD rats were randomly divided into sham group, model group, rt-PA group, rt-PA-Huatan Tongluo group (Chinese medicine group for short). Own embolus method was used to establish rat models of middle cerebral artery embolism (MCAO). The rats in rt-PA group were injected with rt-PA (concentration of 5.67 mg·kg^-1), and those of Chinese medicine group were given with rt-PA and Chinese medicine Huatan Tongluo formulae (7.2 g·kg^-1), 2 times a day. RT-PCR was used in four time phases of 6 hours, 1 day, 3, 7 days to detect the expression of GADD153/CHOP and JNK1 mRNA in cerebral cortex infarction tissues in rats. TUNEL method was used to detect brain neurons apoptosis in rats. Result: Compared with sham group, the gene expression of GADD153/CHOP and JNK1 in 4 time phases was significantly higher in model group (P<0.05). Compared with sham group within the same time phase, neurons apoptosis in model group was significantly increased (P<0.05). Compared with the model group, the expression of JNK1 mRNA was decreased in 4 time phases in both rt-PA group and Chinese medicine group (P<0.05), and rt-PA group could decrease GADD153/CHOP mRNA expression on 1 d while Chinese medicine group could decrease the expression on 6 h and 1 d, with significant difference (P<0.05). Neurons apoptosis in rt-PA group on 1 and 7 d was significantly lower than that in the model group (P<0.05), while Chinese medicine group decreased neurons apoptosis in various time phases (P<0.05). However, there was no significant difference between rt-PA group and Chinese medicine group. Conclusion: Huatan Tongluo formulae can prevent and treat partially reperfusion injury after acute cerebra accepted thrombolysis by reducing GADD153/CHOP and JNK1 mRNA expression in apoptosis pathway of endoplasmic reticulum stress, and partially inhibiting the apoptosis of nerve cells.