加味涤痰汤对脑缺血再灌注损伤大鼠脑细胞自噬的特异性

目的探讨加味涤痰汤对脑缺血再灌注(CIR)损伤大鼠脑细胞自噬的特异性。方法选取SPF级雄性SD大鼠90只,分为假手术组、模型组、中药高剂量组、中药低剂量组、西药组。对大鼠进行可逆性脑中动脉闭塞线栓法栓塞右侧大脑中动脉,构建CIR损伤模型,造模成功后24 h,假手术组与模型组灌胃生理盐水,中药高剂量组、中药低剂量组分别灌胃加味涤痰汤0.768 g/(kg·d)、0.384 g/(kg·d),西药组灌胃吡拉西坦片0.5 g/(kg·d),灌胃容积为10 ml/kg;1次/d,连续给药7 d。于给药周期结束24 h内,每组取5只大鼠行Western印迹测定脑组织中自噬相关蛋白微管相关蛋白1轻链(LC)3-Ⅱ、Beclin1、Bcl-2的表达。结果与假手术组比较,模型组缺血脑组织中LC3-Ⅱ、Bcl-2表达显著升高。与模型组比较,中药高剂量组、低剂量组及西药组LC3-Ⅱ表达均显著降低,3组LC3-Ⅱ/β-actin、Bcl-2/β-actin比值与模型组比较差异有统计学意义(P<0.05)。与假手术组比较,模型组缺血脑组织中Beclin1表达显著升高,与模型组比较,中药高剂量组及西药组Beclin1表达显著下降,两组Beclin1/β-actin比值与模型组比较差异有统计学意义(P<0.05)。结论加味涤痰汤能干预CIR损伤后的自噬,调节自噬的活性,抑制LC3-Ⅱ、Beclin1的表达,有效下调自噬,起到神经保护作用。

The specificity of modified Di tan modified decoction on autophagy in brain cells of rats with cerebral ischemia reperfusion injury

Objective To investigate the effect of modified Di tan modified decoction on autophagy in brain cells of rats with cerebral ischemia reperfusion injury.Methods 90 male SD rats of grade SPF were selected and divided into 5 groups:sham,model,high,low dose of traditional Chinese medicine groups and Western medicine group.Right middle cerebral occlusion was made by reversible middle cerebral artery occlusion suture-occluded method to make construction of reperfusion(CIR)injury model.After the success of the model 24 h,sham operation group and model group were lavaged with physiological saline,high and low dose of traditional Chinese medicine groups were orally modified Di tan modified decoction 0.768 g/(kg·d)and 0.384 g/(kg·d),Western medicine group was intragastric administration of Piracetam tablets 0.5 g/(kg·d),intragastric volume 10 ml/kg,once a day.Medicine was continuously given for 7 days.Within 24 h,5 rats in each group were subjected to Western blot to determine the expressions of autophagy related protein LC3-Ⅱ,Beclin1 and Bcl-2 in brain tissues at the end of the administration cycle.Results Compared with sham group,the expressions of LC3-Ⅱ and Bcl-2 in the ischemic brain tissue of model group were significantly increased(P<0.05).Compared with model group,expression of LC3-Ⅱ was significantly decreased in high dose of traditional Chinese medicine group,low dose of traditional Chinese medicine group and Western medicine group(P<0.05).Compared with sham group,the expression of Beclin1 in brain tissue of model group was significantly increased(P<0.05).Compared with model group,the expression of Beclin1 was significantly decreased in high dose of traditional Chinese medicine group and Western medicine group(P<0.05).Conclusions Di tan modified decoction could interfere with autophagy after MCAO/R,regulate the activity of autophagy,inhibit the expression of LC3-Ⅱ and Beclin1,effectively down regulate autophagy,and play a neuroprotective role after ischemia reperfusion.