强制性运动疗法及“白脉散”有效成分组对实验性MCAO/R模型神经血管单元细胞凋亡的影响及机制研究

Mechanisms of Constraint-induced Movement Therapy and Effective Compounds Group of “Baimai” Powder on the Apoptosis Cells of Neurovascular Unit on Experimental MCAO/R Model

目的:研究强制性运动疗法、"白脉散"抗脑缺血有效成分组及二者联用对大鼠脑缺血再灌注损伤的保护作用及相关机制。方法:75只雄性SD大鼠随机分为假手术组(Sham)、模型组(MCAO/R)、银杏叶提取物组(EGB)、强制性运动疗法组(CIMT)、"白脉散"有效成分组(ECG),强制性运动疗法+"白脉散"有效成分组(CIMT+ECG),每组7只。采用线栓法制作大鼠大脑中动脉闭塞再灌注模型,缺血2h再灌注24h后进行神经功能评分,然后ECG组及CIMT+ECG组以155mg/kg/d灌胃,EGB组以5.6 mg/kg/d灌胃,其余组灌胃等体积的溶媒,同时CIMT组及CIMT+ECG组建立运动疗法模型,每天1次,连续7天。利用免疫组化法观察神经血管单元(NVU)细胞中Bcl-2、Caspase-3蛋白表达的情况,运用原位末端标记(TUNEL)法计数NVU凋亡细胞。结果:CIMT与"白脉散"ECG单独或联合应用可显著改善模型大鼠神经功能评分、抑制NVU细胞凋亡、增加Bcl-2蛋白的表达,降低Caspase-3蛋白的表达。结论:CIMT及"白脉散"ECG可保护MCAO/R模型大鼠神经功能,其机制可能与其抗NVU细胞凋亡,调节凋亡调控因子蛋白表达有关,且二者联用比二者单用具有更佳的保护效果。

Objective： To study the neuroprotective effects and related mechanisms of constraint -induced movement therapy（ CI- MT）, effective compounds group （ECG） of ＂Baimai Powder＂ and combination of the two on cerebral ischemia- reperfusion inju- ry in rats. Method： 75 adult Sprague Dawley rats were randomly divided into 6 groups： sham- operated group （Sham）, middle cerebral artery occlusion/reperfusion group（ MCAO/R）, extract of Ginkgo Biloba group （EGB）, constraint - induced movement therapy group （CIMT） ,＂ Baimai Powder＂ ECG group （ECG）, constraint -induced movement therapy ＋ ＂Baimai Powder＂ ECG group（CIMT ＋ ECG）. The model of middle cerebral artery occlusion（MCAO） was established by thread ligation method, ische- mia 2h then reperfusion 24h, then the neurological score was taken , CIMT ＋ ECG group and ECG group were ig with the dosage of 155 mg/kg/d, EGB group was ig with the dosage of 5.6 mg/kg/d, the equivalent vehicle was given to the other groups, all groups were ig for 7d, respectively, Sports rehabilitation model was established to the CIMT group and CIMT ＋ ECG group at the same time. Then the protein expression of Bcl - 2 and Caspase - 3 of the cells of the neurovascular unit （NVU） were detected by immuno - hisochemistry, apoptosis cells of the NVU were detected by means of TdT - Mediated dUTP nick end labeling （TUNEL）. Result： CIMT and ECG of ＂Baimai Powder＂ singly or in combination can significantly improve the neurological score, restrain the apoptosis cells of the NVU, increase the expression of Bcl - 2 positive cells and decrease the expression of Caspase -3 positive cells. Conclusion： CIMT and ECG of ＂Baimai Powder＂ can remarkably improve the neural behavioral func- tions of model rats by resisting the apoptosis cells of the NVU and adjusting protein expression of apoptosis - regulating cyto- kines. The curative effect of the combination of CIMT and the ECG of ＂Baimai Powder＂ is much better.