中文摘要：

目的：基于Calpain/p35-Cdk5/p25通路探究肾脑复元汤联合人脐带间充质干细胞（h UC-MSCs）干预对缺氧缺糖神经元的保护作用。方法：原代培养神经元,并随机分为正常组、模型组（缺糖缺氧造模,OGD）、中药组（OGD＋含中药血清）、干细胞组（OGD＋h UC-MSCs共培养）、联合组（OGD＋含中药血清＋h UC-MSCs共培养）、抑制剂组（OGD＋Calpain抑制剂）,采用CCK-8法检测神经元活力,AO/EB染色观察细胞形态学变化,流式细胞仪检测各组细胞凋亡率,Werstern blot法检测Cdk5、p25、p35的表达。结果：模型组神经元缺糖缺氧后,突起回缩,轴突网络稀释,细胞核变性、甚至碎裂,与正常组比较,细胞活力明显下降,大量细胞凋亡,Cdk5、p25表达明显上升,p35表达明显下降（P〈0.01）;与模型组比较,中药组、干细胞组、联合组及抑制剂组神经元活力升高,凋亡细胞数降低,Cdk5及p25表达下降,p35表达上升（P〈0.05,P〈0.01）;与中药组、干细胞组及抑制剂组相比,联合组神经元活力升高、早凋细胞减少（P〈0.05）;联合组及抑制剂组CDK5、p25表达较中药组及干细胞组下降,p35表达较中药组及干细胞组上升（P〈0.05）。结论：肾脑复元汤联合h UC-MSCs移植能显著改善缺糖缺氧神经元凋亡,其作用机制可能与调控Calpain/p35-Cdk5/p25通路有关。

英文摘要：

Objective： To investigate the neuroprotective effects of combination of Shennao Fuyuan Decoction and human umbilical cord mesenchymal stem cells（h UC-MSCs） on oxygen-glucose deprive（OGD） neurons of ischemia stroke based on Calpain/p35-Cdk5/p25 signal pathway. Methods： Primary cultured neurons were randomly divided into normal group, model group, TCM group（OGD＋drug serum of TCM）, stem cell group（OGD＋h UC-MSCs co-culture）, combination group（OGD ＋ medicated serum of TCM＋h UC-MSCs administration）, inhibitor group（OGD＋inhibitor of calpain）. The neuron vitality was evaluated by CCK8, the pathological changes of neurons were observed by AO/EB staining, apoptotic neurons were detected by FCM, the expression of CDK5, p25, p35 was detected by Werstern-blot method. Results： After OGD, axones of model group retracted and became sparse, nucleus degenerated or became cataclastic, cell vitality decreased apparently, the expression of Cdk5, p25 increased while p35 expression decreased（P〈0.05, P〈0.01）. Compared with TCM, stem cell and inhibitor group, cell vitality of neurons in combination group was higher, apoptotic neurons were less（P〈0.05）. The expression of Cdk5 and p25 in combination and inhibitor group were lower than TCM and stem cell group, p35 expression was higher than TCM and stem cell group（P〈0.05）. Conclusion： The combination of Shennao Fuyuan Decoction and h UC-MSCs implantation could significantly reduce the apoptosis of neurons after OGD, this effect was probably due to the regulation of Calpain/p35-Cdk5/p25 pathway.