神经干细胞过表达Hsp75降低Aβ介导的神经毒性

发布时间：2018-07-09 20:21

本文选题：热休克蛋白 + C.神经干细胞　；参考：《中国病理生理杂志》2016年02期

【摘要】：目的:应用腺病毒为载体在体外过表达热休克蛋白75(Hsp75),研究Hsp75蛋白过表达对神经干细胞在Aβ诱导神经毒性中的作用,并初步探讨其作用机制。方法:体外培养小鼠神经干细胞C17.2,实验分为对照组、Aβ处理组、腺病毒阴性感染组和腺病毒Hsp75过表达感染组。使用荧光显微镜观察腺病毒的感染情况并进行细胞免疫鉴定;倒置相差显微镜观察各组神经干细胞的形态;MTT法检测细胞活力;流式细胞术检测细胞凋亡率;Western blot检测Hsp75和活化型caspase-3蛋白水平。结果:荧光显微镜观察和Western blot检测表明腺病毒成功感染神经干细胞并高效表达Hsp75蛋白。此外,腺病毒感染不会导致细胞形态改变及细胞分化,同时也不会影响细胞活力。与对照组比较,Aβ处理组及腺病毒阴性感染组细胞存活率显著降低(P0.05),细胞凋亡率及活化型caspase-3蛋白水平显著增高(P0.05);然而,Hsp75过表达能显著提高神经干细胞的存活率,减少细胞凋亡和降低活化型caspase-3蛋白水平(P0.05)。结论:Hsp75过表达对Aβ诱导损伤的C17.2细胞具有明显保护作用,其机制可能是与抑制caspase-3途径依赖的细胞凋亡有关。
[Abstract]:Aim: to investigate the effect of overexpression of Hsp75 protein on neural stem cells induced by A 尾 -induced neurotoxicity in vitro using adenovirus as a vector to overexpression heat shock protein 75 (Hsp75), and to explore the mechanism of the overexpression of Hsp75 protein. Methods: C17.2mouse neural stem cells were cultured in vitro and divided into control group, adenovirus negative infection group and adenovirus Hsp75 overexpression group. The infection of adenovirus was observed by fluorescence microscope, and the cell viability was detected by MTT assay with inverted phase contrast microscope. Apoptosis rate was detected by flow cytometry and the levels of Hsp75 and activated caspase-3 protein were detected by Western blot. Results: fluorescence microscopy and Western blot analysis showed that adenovirus successfully infected neural stem cells and expressed Hsp75 protein. In addition, adenovirus infection does not cause cell morphological changes and cell differentiation, nor does it affect cell viability. Compared with the control group, the cell survival rate decreased significantly (P0.05), the apoptosis rate and the level of activated caspase-3 protein increased significantly in the A 尾 treated group and the adenovirus negative infection group (P0.05), but the overexpression of Hsp75 significantly increased the survival rate of neural stem cells. To reduce apoptosis and reduce the level of activated caspase-3 protein (P0.05). Conclusion the overexpression of 1: Hsp75 has a significant protective effect on A 尾 -induced C17.2 cells, and the mechanism may be related to the inhibition of apoptosis dependent on the caspase-3 pathway.
【作者单位】：南方医科大学珠江医院药剂科;广东省中西医结合医院药事管理科;中山大学附属第三医院肝胆外科;广州市中西医结合医院药剂科;南方医科大学珠江医院烧伤科;
【基金】：广东省自然科学基金资助项目(No.S2012010008199;No.S2013010015546)
【分类号】：R749.16

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