敲除TRPC6对MPTP诱导小鼠神经炎性损伤的影响

发布时间：2018-08-31 09:48

【摘要】：目的在小鼠MPTP(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine)诱导的神经炎症模型中检测小胶质细胞的TRPC6通道的表达,并探究TRPC6通道对于炎症表达和多巴胺能神经元损伤的影响。方法在MPTP注射的小鼠中,用标记CD11b的磁珠分选出黑质区域的小胶质细胞,并通过蛋白质免疫印迹的方法检测TRPC6的表达。在构建CD11b-TRPC6~(-/-)小鼠后,通过免疫荧光的方法检测MPTP诱导的小胶质细胞的增殖以及多巴胺能神经元的损伤。同时结合蛋白质免疫印迹和实时荧光定量PCR的方法,在MPTP模型中检测TRPC6敲除后cryαB蛋白水平和炎症因子表达。结果在MPTP注射的小鼠中,小胶质细胞中TRPC6通道的表达显著上调。另外,在注射MPTP的CD11b-TRPC6~(-/-)小鼠中,小胶质细胞中的cryαB蛋白水平被上调。同时,MPTP诱导的小胶质细胞的增殖以及炎症因子表达增强被抑制,而多巴胺能神经元的损伤得到缓解。结论小胶质细胞中TRPC6通道表达在MPTP模型中上调,而TRPC6的敲除可增加小胶质细胞中cryαB蛋白水平,并降低炎症因子的表达,从而减少多巴胺能神经元的损伤。
[Abstract]:Objective to detect the expression of TRPC6 channel in microglia in mouse MPTP (1-methyl-4-phenyl-1aniline) induced neuritis model, and to explore the effect of TRPC6 channel on inflammatory expression and dopaminergic neuron injury. Methods microglia in substantia nigra were isolated from mice injected with MPTP and the expression of TRPC6 was detected by Western blot. After constructing CD11b-TRPC6~ (- / -) mice, the proliferation of microglia induced by MPTP and the damage of dopaminergic neurons were detected by immunofluorescence. The level of cry 伪 B protein and the expression of inflammatory factors after TRPC6 knockout were detected in MPTP model by Western blot and real-time fluorescent quantitative PCR. Results in mice injected with MPTP, the expression of TRPC6 channel in microglia was significantly up-regulated. In addition, the level of cry 伪 B protein in microglia was up-regulated in CD11b-TRPC6~ (-r -) mice injected with MPTP. At the same time, MPTP induced proliferation of microglia and increased expression of inflammatory factors were inhibited, while the damage of dopaminergic neurons was alleviated. Conclusion the expression of TRPC6 channel in microglia is up-regulated in MPTP model, and the knockout of TRPC6 can increase the level of cry 伪 B protein in microglia cells and decrease the expression of inflammatory factors, thus reducing the damage of dopaminergic neurons.
【作者单位】：漳州卫生职业学院;厦门大学附属东南医院(福建省漳州中国人民解放军第175医院);
【基金】：国家自然基金(81601223)
【分类号】：R742.5

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