基质衍生因子-1对大鼠脑出血后神经细胞凋亡的作用及其机制研究  

Mechanism of stromal derived factor(SDF-1) on nerve cells apoptosis after hemorrhagic brain injury in rat

作　　者：黎柏源[1] 白万胜[1] 赵永博[1] 孙平安[1] 周波[1] 胡彪[1] 

机构地区：[1]解放军第323医院神经外科,陕西西安710054

出　　处：《中国现代医学杂志》2015年第1期21-25,共5页China Journal of Modern Medicine

摘　　要：目的研究基质衍生因子-1(SDF-1)对大鼠脑出血后神经细胞凋亡的作用及其可能机制,从而为探讨SDF-1/CXCR4信号通路在脑出血后继发损伤中的作用奠定基础。方法健康雄性SD大鼠30只,利用立体定位技术向大鼠尾状核内注入VII型胶原酶0.5U建立脑出血模型。设立假手术组、生理盐水对照组、SDF-1处理组,各组于术后72 h经尾状核行冠状切片,TUNEL染色观察神经细胞凋亡,RT-PCR和Western-Blot测定SDF-1的m RNA和蛋白水平。建立体外诱导大鼠皮质神经元凋亡模型,给予SDF-1及其受体CXCR4抑制剂AMD3100处理,流式细胞仪检测神经细胞凋亡情况,Western-Blot法测定凋亡相关蛋白caspase-3基因蛋白表达。结果与假手术组和对照组相比,SDF-1处理组中SDF-1的m RNA和蛋白水平明显增多;且血肿周围凋亡细胞数低于对照组。体外分析表明,80 ng/ml SDF-1处理后可明显减少神经细胞的凋亡,同时降低凋亡相关蛋白caspase-3蛋白的表达水平;AMD3100预处理后,SDF-1抗凋亡能力明显下降,同时caspase-3的水平无明显变化。结论 SDF-1具有抗神经细胞凋亡作用,这一作用可能是由CXCR4介导caspase-3通路来实现的,提示SDF-1/CXCR4可作为受损后神经保护的潜在靶点。[Objective] To study the possible mechanism of stromal derived factor -1 （SDF-1） on nerve cells apoptosis after hemorrhagic brain injury, so as to explore the SDF-1/CXCR4 axis as a foundation in secondary injury after intracerebral hemorrhage. [Methods] 30 Male Sprague-Dawley rats were used to induce an ICH model. Sham group, control group and SDF-1 group were treated respectively, 72 hours after the operation, brains were cut into slices. The nerve cells apoptosis was analyzed by TUNEL assay, mRNA and protein levels of SDF-1 were determined by RT-PCR and Western-Blotting. Furthermore, the model of cortical neurons apoptosis in vitro was established, stimulated with certain concentration of SDF-1, then nerve cells apoptosis was analyzed by flow cytometry, the expression levels of easpase-3 mRNA and protein were detected by western blotting after pretreatment with the CXCR4 inhibitor AMD3100. [Results] mRNA and protein level of SDF-1 in rats with hemorrhagic brain injury were significantly increased, and the number of apoptotic ceils was lower than the control group; further mechanism analysis showed that SDF-1 reduced the apoptosis of neural cells, and decreased the expression of apoptosis related protein caspase-3, which can be significantly abolished by pretreatment withCXCR4 inhibitor AMD3100. [Conclusion] SDF-1 can reduce the apoptosis of neural ceils after hemorrhagic brain injury in rat CXCR4-mediated caspase-3 pathway. Consequently, our findings could clarify a new explanation about how SDF-1 reduces the apoptosis of neurons and provide a potential target for neuron protection after damage.

关 键 词：SDF-1/CXCR4 脑出血 神经细胞 凋亡 

分 类 号：R743[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]