TRPC1沉默对脑缺血大鼠神经干细胞迁移的影响

发布时间：2018-12-13 11:26

【摘要】：目的探讨瞬时受体电位通道1(TRPC1)沉默对脑缺血大鼠内源性神经干细胞增殖、迁移和分化的影响。方法 SD大鼠分为假手术组、脑缺血组、TRPC1沉默组(脑缺血+TRPC1沉默)和阴性对照组,以脑室内注射siRNA沉默TRPC1,以线栓法制作大鼠大脑中动脉脑缺血(MCAO)模型,脑缺血模型制作成功后,腹腔内注射Brdu标记内源性神经干细胞,分别于48 h、4 w后处死大鼠,行Brdu免疫组化染色、免疫荧光双标染色(Brdu/GFAP、Brdu/Neun)观察NSC的增殖、迁移和分化情况。结果脑缺血后48 h,脑缺血组和TRPC1沉默组SVZ区Brdu阳性表达均较假手术组明显增多(P0.01),但TRPC1沉默组Brdu阳性细胞数较缺血组低(P0.01)。脑缺血4 w后,缺血组与假手术组相比,皮质区具有更多的Brdu阳性细胞(P0.01),同样TRPC1沉默组Brdu阳性细胞数多于假手术组,但明显低于脑缺血组(P0.01);免疫荧光双标染色发现,脑缺血各组Brdu/GFAP、Brdu/Neun双阳性细胞的数量均比假手术组明显增高,但TRPC沉默组双阳性细胞显著少于脑缺血组和阴性对照组(P0.01)。结论 TRPC1沉默显著影响脑缺血大鼠SVZ区NSC的增殖、向脑缺血区迁移及向成熟细胞的分化。
[Abstract]:Objective to investigate the effects of transient receptor potential channel 1 (TRPC1) silencing on the proliferation, migration and differentiation of endogenous neural stem cells (NSCs) in rats with cerebral ischemia. Methods SD rats were divided into three groups: sham operation group, cerebral ischemia group, TRPC1 silencing group (cerebral ischemia TRPC1 silencing) and negative control group. The rat middle cerebral artery ischemic (MCAO) model was made by intraventricular injection of siRNA silencing TRPC1, with thread embolization. After the model of cerebral ischemia was established successfully, Brdu was injected intraperitoneally to label endogenous neural stem cells. The rats were killed at 48 h or 4 weeks later. The proliferation of NSC was observed by Brdu immunohistochemical staining and double immunofluorescence staining (Brdu/GFAP,Brdu/Neun). Migration and differentiation. Results at 48 hours after cerebral ischemia, the positive expression of Brdu in SVZ in cerebral ischemia group and TRPC1 silencing group was significantly higher than that in sham operation group (P0.01), but the number of Brdu positive cells in TRPC1 silencing group was lower than that in ischemic group (P0.01). After 4 weeks of cerebral ischemia, there were more Brdu positive cells in the cortex in the ischemic group than in the sham operation group (P0.01). The number of Brdu positive cells in the same TRPC1 silencing group was higher than that in the sham operation group, but significantly lower than that in the ischemic group (P0.01). Immunofluorescence double labeling staining showed that the number of Brdu/GFAP,Brdu/Neun double positive cells in cerebral ischemia group was significantly higher than that in sham operation group, but the number of double positive cells in TRPC silencing group was significantly lower than that in cerebral ischemia group and negative control group (P0.01). Conclusion TRPC1 silencing has a significant effect on the proliferation of NSC in the SVZ area, migration to the ischemic area and differentiation into mature cells of cerebral ischemia rats.
【作者单位】：连云港市第一人民医院神经内科;重庆医科大学附属第一医院神经内科;
【基金】：中国博士后科学基金(No.2016M601759) 连云港市第一人民医院博士科研启动基金
【分类号】：R743.3

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