化痰通络方对急性脑梗死大鼠rt-PA 溶栓后神经细胞自噬途径中Beclin1 与LC3 蛋白表达的影响

Effects of Huatan Tongluo formulae on expression of Beclin1 and LC3 on gene neuronal autophagy pathway in rat cortex accepted thrombolysis therapy after acute cerebral infarction

关键词:  化痰通络方  大鼠重组组织纤溶酶原激活剂  溶栓  急性脑梗死  自噬途径  自噬相关蛋白  微管相关蛋白轻链

Key words:  Huatan Tongluo formulae  rt-PA  thrombolysis  acute cerebral infarction  autophagy pathway  Beclin1  LC3

摘要:

[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)溶栓后神经细胞自噬途径中自噬相关蛋白(Beclin1)及微管相关蛋白轻链3(LC3)蛋白表达的影响。[方法] 选择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),每组采用6 h、1 d、3 d和7 d 4个时相。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(浓度为5.67 mg/kg)及联合化痰通络中药(浓度为7.2 g/kg)干预,每日2 次。于相应时间点采用Western blot 检测各组大鼠大脑皮质梗死区组织中Beclin1及LC3 蛋白的表达。[结果] Western blot结果显示:与假手术组相比,模型组在4 个时相中Beclin1和LC3 蛋白的相对丰度值均明显增高(P<0.05);采用rt-PA 和rt-PA联合化痰通络法干预后,4 个时相内Beclin1与LC3 蛋白的相对丰度值均明显下降(P<0.05);且rt-PA 联合化痰通络组下降更加显著,与rt-PA 组比较差异有统计学意义(P<0.05)。[结论] 化痰通络方可通过降低神经细胞自噬途径中Beclin1 与LC3 蛋白的表达,部分抑制神经细胞的过度自噬,进而保护神经细胞损伤,从而更好的防治急性脑梗死溶栓后缺血再灌注的发生与发展。

Abstract:

[Objective] To observe the effects of Huatan Tongluo formulae on expression of Beclin1 and LC3 on gene neuronal autophagy pathway in rat cortex accepted thrombolysis therapy after the acute cerebral infarction. [Methods] 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), each group was divided into four time phases: 6 hours, 1 day, 3 days and 7 days. In the corresponding time points prepares the MCAO rat model, the rats in rt-PA group were injected rt-PA(concentration of 5.67 g/kg), and those of Chinese medicine group were given rt-PA and Chinese medicine Huatan Tongluo, 2 times a day. Then respectively observe the protein expression of Beclin1 and LC3 by Western Blot. [Results] The results of Western Blot showed: compared with sham group, the expression of Beclin1 and LC3 in model group increased significantly(P<0.05), compared with model group in the same time phases, the expression of Beclin1 and LC3 decreased in rt-PA group and Chinese medicine group(P<0.05),especially in Chinese medicine group. [Conclusion] Huatan Tongluo formulae can inhibit the activation of Beclin1 and LC3 on gene neuronal autophagy pathway, and inhibit the nerve cells excessive autophagy at a certain extent, to better prevention and treatment of reperfusion injury after acute cerebra accepted thrombolysis.