黄芪皂甙Ⅳ联合BMSCs移植对大鼠脑缺血再灌注海马神经细胞凋亡及Livin、Caspase-9蛋白的影响

目的探讨黄芪皂甙Ⅳ联合骨髓间充质干细胞（BMSCs）移植对缺血再灌注大鼠海马Livin、Caspase-9蛋白及神经细胞凋亡表达的影响。方法实验动物随机分为假手术组、模型组、BMSCs组、联合组。采用线栓法建立大鼠大脑中动脉缺血（MCAO）模型,假手术组不予处理。BMSCs组于造模后3 h移植BMSCs。联合组既移植BMSCs,又给予黄芪皂甙Ⅳ治疗。观察缺血72 h后,各组神经功能评分,PKH-26标记的移植BMSCs分布,采用Western blot和免疫组化方法检测大鼠海马Livin、Caspase-9蛋白的表达,TUNEL方法检测细胞凋亡指数。结果 PKH-26标记的BMSCs在海马有表达。各组中联合组神经功能恢复最为明显（P〈0.05）。与模型组比较,联合组和BMSCs组均可上调Livin蛋白表达（P〈0.05）,下调Caspase-9蛋白表达（P〈0.05）,降低神经元细胞凋亡指数（P〈0.05）,以联合组作用更为显著（P〈0.05）。结论黄芪皂甙Ⅳ联合BMSCs移植对脑缺血再灌注神经元细胞凋亡有协同抑制作用,其作用机制可能与黄芪皂甙Ⅳ促进BMSCs上调Livin蛋白表达,下调Caspase-9蛋白表达,抑制细胞凋亡有关。

Influence of Astragaloside ⅣCombined with Bone Mesenchymal Stem Cell Transplantation on Expressions of Livin,Caspase-9 and Neural Cell Apoptosis in Hippocampus after Cerebral Ischemia-reperfusion in Rats

Objective To investigate the effects of Astragaloside Ⅳ combined with bone mesenchymal stem cell（BMSCs） on the apoptosis of neural cells and expressions of livin and caspase-9 in hippocampal region after cerebral ischemia-reperfusion in rats.Methods A total of 42 healthy SD rats were randomly divided into sham operation group,model group,BMSCs group,and combination group.The rat models with middle cerebral artery occlusion were established.The models in BMSCs and combination groups were treated with suture method,and the rats in four groups were injected with PBS and PKH-26 labeled BMSCs suspension via caudal vein 3 hours after middle cerebral artery occlusion（MCAO）respectively.Astragaloside Ⅳ（25 mg/kg） was administered in combination group.After reperfusion 72h,neurological function scores were observed.The expressions of livin and caspase-9 proteins were determined by immunohistochemical method and Western blot.The apoptosis of neural cells was detected by TUNEL method.Results PKH-26 labeled BMSCs were observed in hippocampus.Compared with model and BMSCs group,the neurological scores in combination group were decreased significantly（P〈0.05）.Compared with model group,the number of apoptotic neural cells and the expression in caspase-9 proteins were significantly lower in BMSCs group and combination group（P〈0.05）.The expression of livin proteins was significantly increased（P〈0.05）.The effect in combination group was more powerful than that in BMSCs group（P〈0.05）.Conclusion The combination group could play coordinate repression on neural cell apoptosis in rats after cerebral ischemia-reperfusion,which might be related to astragaloside Ⅳ to improve BMSCs on up-regulating livin expression,down-regulating Caspase-9 expression.