钙敏感受体对海马神经元细胞缺氧复氧损伤的作用

发布时间：2018-04-25 10:09

本文选题：钙敏感受体 + 缺氧/复氧　；参考：《青岛大学》2016年博士论文

【摘要】：目的:探讨钙敏感受体(Ca SR)在缺氧复氧条件下对海马神经元细胞的作用及其可能的机制。方法:利用原代培养的新生大鼠脑内的海马神经元细胞建立缺氧复氧损伤模型,采用随机数字表法分成四组,分别为正常对照组、缺氧复氧组(H/R)、激动剂处理(Gd Cl_3)组、拮抗剂处理(NPS-2390)组。在倒置显微镜下观察海马神经元细胞数量和细胞形态,同时,利用MTT法检测神经元细胞的存活情况。应用流式细胞仪检测四个组的神经元细胞的凋亡率,应用Western-blot法检测神经元细胞中Ca SR,caspase-3,Bcl-2蛋白表达和细胞色素(Cyt C)释放,同样应用Western-blot法检测ERK和P38蛋白磷酸化水平。结果:与对照组比较,H/R组的细胞数目显著减少,与H/R组比较,Gd Cl_3组细胞数量进一步减少,而钙敏感受体拮抗剂组的细胞数目有效增加。与对照组比较,H/R组及激动剂处理(Gd Cl_3)组中神经元细胞凋亡率明显升高,细胞凋亡率在拮抗剂(NPS-2390)组中显著下降(P0.05)。与对照组比较,H/R组及激动剂处理(Gd Cl_3)组细胞中caspase-3,BAX蛋白表达上调和细胞色素(Cyt C)释放增多,与上两组比较,拮抗剂(NPS-2390)组caspase-3,BAX蛋白表达减少和细胞色素(Cyt C)释放减少(P0.05)。在四组中,ERK1/2,P38和p P38的表达没有显著差异,但是p ERK1/2的表达在H/R组及激动剂处理(Gd Cl_3)组显著增加,在拮抗剂(NPS-2390)组减少(P0.05)。结论:抑制钙敏感受体的表达能够有效减轻海马神经元的缺血再灌注损伤,抑制缺血再灌注后神经元细胞凋亡,其机制与抑制ERK1/2的磷酸化有关。
[Abstract]:Aim: to investigate the effect of calcium sensitive receptor (Ca SRR) on hippocampal neuronal cells under hypoxia reoxygenation and its possible mechanism. Methods: the model of hypoxic reoxygenation injury was established by primary cultured hippocampal neurons in neonatal rats. The model was randomly divided into four groups: normal control group, hypoxia reoxygenation group (H / R) group, and agonist treatment group (GdCl _ 3). Antagonists treated with NPS-2390) group. The number and morphology of hippocampal neurons were observed under inverted microscope, and the survival of neurons was detected by MTT method. The apoptotic rate of neurons in four groups was detected by flow cytometry, the expression of Ca SR-caspase-3 Bcl-2 protein and cytochrome Cyt C release were detected by Western-blot assay, and the phosphorylation levels of ERK and P38 protein were also detected by Western-blot method. Results: compared with the control group, the number of cells in the H / R group was significantly decreased, and that in the Gd Cl_3 group was further decreased, while that in the calcium sensitive receptor antagonist group was significantly increased. Compared with the control group, the apoptosis rate of neurons in the H / R group and the agonist treated GdCl _ 3 group was significantly increased, and the apoptosis rate in the antagonist NPS-2390 group was significantly decreased (P 0.05). Compared with the control group, the expression of caspase-3mBAX protein and the release of cytochrome Cyt C increased in the H- / R group and the agonist treated GdCl _ 3 group. Compared with the above two groups, the expression of caspase-3mBAX protein and the cytochrome cyt C release decreased in the antagonist NPS-2390 group. There was no significant difference in the expression of P38 and pP38 between the four groups, but the expression of p ERK1/2 was significantly increased in the H / R group and the agonist treated GdCl _ 3 group, but decreased in the antagonist group (NPS-2390). Conclusion: inhibiting the expression of Ca ~ (2 +) sensitive receptor can effectively reduce the ischemia-reperfusion injury of hippocampal neurons and inhibit the apoptosis of neurons after ischemia-reperfusion. The mechanism is related to the inhibition of phosphorylation of ERK1/2.
【学位授予单位】：青岛大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R743.3

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