骨髓间充质干细胞外泌体对氯化钴引起的神经干细胞凋亡的影响

Effect of Exosomes Derived from Mesenchymal Stem Cells on Apoptosis of Neural Stem Cells Induced by Cobalt Chloride

目的探讨鼠骨髓间充质干细胞外泌体对神经干细胞凋亡及Wnt信号通路的影响。方法提取大鼠原代骨髓间充质干细胞,收集细胞上清后超速离心分离得到外泌体并予以鉴定。通过加入氯化钴(CoCl2)诱导神经干细胞发生化学性缺氧,从而导致细胞凋亡,CCK8检测氯化钴引起神经干细胞凋亡的最适浓度。对神经干细胞分为空白对照组、氯化钴组、外泌体组,培养12h通过Tunel检测细胞凋亡,并收集细胞Westernblot检测凋亡标志蛋白、凋亡相关蛋白及Wnt信号通路蛋白表达。结果通过Tunel可以看出外泌体能够降低神经干细胞的凋亡,并且能够通过抑制DNA修复酶(PARP)的切割,减轻神经干细胞的凋亡;并且能上调B淋巴细胞瘤/白血病-2蛋白(Bcl-2)和下调Bcl-2相关x蛋白(Bax)的表达;同时能够减轻氯化钴导致的Wnt信号通路的激活。结论骨髓间充质干细胞外泌体能够通过抑制Wnt信号通路抑制氯化钴对神经干细胞发生的凋亡诱导作用。

Objective To investigate the effect of exosomes derived from rat mesenchymal stem cells on the apoptosis of neural stem cell and Wnt signaling pathway.Methods Primary mesenchymal stem cells of rats were extracted,and the supernatant was collected and ultracentrifuged,and then exosomes were obtained;chemical hypoxia was induced in neural stem cells by adding cobalt chloride,which led to apoptosis;the optimal concentration of cobalt chloride to induce apoptosis was detected by CCK8;the neural stem cells were divided into 3 groups:blank control group,cobalt chloride group and exosome group;the cells were cultured for 12 hours,and cell apoptosis was detected by Tunel and the expression of apoptotic marker protein,apoptosis-related protein and Wnt signaling pathway by Western blot.Results The exosomes reduced the apoptosis of neural stem cells,attenuated the apoptosis through inhabiting the cleavage of PARP,up-regulated the expression of B lymphoma/leukemia-2 protein(Bcl-2),down-regulated the expression of Bcl-2 related x protein(Bax) and alleviated the activation of the Wnt signaling pathway caused by cobalt chloride.Conclusions The exosomes derived from mesenchymal stem cells can inhabit the inducing effect of cobalt chloride on the apoptosis of neural stem cells by inhabiting Wnt signaling pathway.