脑外伤后T型钙通道对神经干细胞增殖的诱导作用

发布时间：2018-06-15 14:31

本文选题：脑外伤 + 神经干细胞　；参考：《西安交通大学学报(医学版)》2016年04期

【摘要】：目的探讨脑外伤后T型钙通道对神经干细胞的诱导作用及其相关机制。方法构建成年大鼠脑损伤模型,分为手术组和手术+米贝地尔组(米贝地尔组),并另设正常对照组、假手术组。分离侧脑室壁的脑室下层(subventricular zone,SVZ)细胞,用神经干细胞悬浮培养法进行培养并鉴定其干细胞性质;在神经干细胞培养基中加入不同剂量的米贝地尔,通过计数神经干细胞球、噻唑蓝(MTT)法分析米贝地尔对神经干细胞增殖抑制作用,计算米贝地尔对神经干细胞的半抑制浓度(IC50),Western blot法检测脑组织中Cav3.2以及神经干细胞中Cav3.2、Cyclin A和caspase-3的蛋白表达。结果体内实验表明,手术组中神经干细胞的增殖能力显著高于正常对照组和假手术组,应用米贝地尔腹腔注射抑制脑组织中T型钙通道后,神经干细胞的增殖明显降低。体外细胞实验表明,米贝地尔能够明显抑制神经干细胞球的形成,MTT法显示米贝地尔浓度大于5μmol/L时,可显著抑制细胞增殖,5、10、20μmol/L组的吸光度(A值)与对照组比较,差异具有统计学意义(P0.05)。IC50值为8.93μmol/L。Western blot检测结果显示,米贝地尔组的Cav3.2蛋白和Cyclin A蛋白的表达显著降低,caspase-3蛋白的表达显著升高。结论脑外伤后可激活脑组织中T型钙通道,上调神经干细胞的增殖能力。
[Abstract]:Objective to investigate the induction effect of T-type calcium channel on neural stem cells after brain injury and its related mechanism. Methods the adult rat brain injury model was established and divided into two groups: the operation group and the operation group (mibedil group), and the normal control group and the sham operation group. The subventricular SVZ cells were isolated from the lateral ventricular wall, cultured with neural stem cell suspension culture method and identified their stem cell properties, different doses of Mibedil were added to the neural stem cell culture medium, and the neural stem cell balls were counted. The inhibitory effect of mibedil on the proliferation of neural stem cells was analyzed by using thiazolyl blue method. The semi-inhibitory concentration of mibedil on neural stem cells was calculated. The protein expression of Cav3.2 and Cav3.2 Cyclin A and caspase-3 in brain tissue and neural stem cells were detected by IC50 blot. Results in vivo experiments showed that the proliferation of neural stem cells in the operation group was significantly higher than that in the normal control group and the sham operation group. The proliferation of neural stem cells was significantly decreased after intraperitoneal injection of Mibedil to inhibit T-type calcium channel in brain tissue. In vitro cell experiments showed that mibedil could significantly inhibit the formation of neural stem cell spheres. MTT assay showed that when the concentration of mibedil was more than 5 渭 mol / L, it could significantly inhibit the cell proliferation and the absorbance of the group with 10 ~ 10 渭 mol / L) compared with the control group. The results of Western blot analysis showed that the expression of Cav3.2 protein and Cyclin A protein in Mibedil group significantly decreased the expression of caspase-3 protein. Conclusion after brain injury, T-type calcium channel can be activated and the proliferation of neural stem cells can be upregulated.
【作者单位】：西安交通大学第二附属医院神经外科;第四军医大学唐都医院神经外科;西安交通大学第二附属医院妇产科;
【基金】：陕西省社会发展攻关项目资助(No.2014K11-01-01-12)~~
【分类号】：R651.15

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