發(fā)布時(shí)間：2018-04-25 10:09

  本文選題：鈣敏感受體 + 缺氧/復(fù)氧��； 參考：《青島大學(xué)》2016年博士論文

【摘要】：目的:探討鈣敏感受體(Ca SR)在缺氧復(fù)氧條件下對(duì)海馬神經(jīng)元細(xì)胞的作用及其可能的機(jī)制。方法:利用原代培養(yǎng)的新生大鼠腦內(nèi)的海馬神經(jīng)元細(xì)胞建立缺氧復(fù)氧損傷模型,采用隨機(jī)數(shù)字表法分成四組,分別為正常對(duì)照組、缺氧復(fù)氧組(H/R)、激動(dòng)劑處理(Gd Cl_3)組、拮抗劑處理(NPS-2390)組。在倒置顯微鏡下觀察海馬神經(jīng)元細(xì)胞數(shù)量和細(xì)胞形態(tài),同時(shí),利用MTT法檢測(cè)神經(jīng)元細(xì)胞的存活情況。應(yīng)用流式細(xì)胞儀檢測(cè)四個(gè)組的神經(jīng)元細(xì)胞的凋亡率,應(yīng)用Western-blot法檢測(cè)神經(jīng)元細(xì)胞中Ca SR,caspase-3,Bcl-2蛋白表達(dá)和細(xì)胞色素(Cyt C)釋放,同樣應(yīng)用Western-blot法檢測(cè)ERK和P38蛋白磷酸化水平。結(jié)果:與對(duì)照組比較,H/R組的細(xì)胞數(shù)目顯著減少,與H/R組比較,Gd Cl_3組細(xì)胞數(shù)量進(jìn)一步減少,而鈣敏感受體拮抗劑組的細(xì)胞數(shù)目有效增加。與對(duì)照組比較,H/R組及激動(dòng)劑處理(Gd Cl_3)組中神經(jīng)元細(xì)胞凋亡率明顯升高,細(xì)胞凋亡率在拮抗劑(NPS-2390)組中顯著下降(P0.05)。與對(duì)照組比較,H/R組及激動(dòng)劑處理(Gd Cl_3)組細(xì)胞中caspase-3,BAX蛋白表達(dá)上調(diào)和細(xì)胞色素(Cyt C)釋放增多,與上兩組比較,拮抗劑(NPS-2390)組caspase-3,BAX蛋白表達(dá)減少和細(xì)胞色素(Cyt C)釋放減少(P0.05)。在四組中,ERK1/2,P38和p P38的表達(dá)沒(méi)有顯著差異,但是p ERK1/2的表達(dá)在H/R組及激動(dòng)劑處理(Gd Cl_3)組顯著增加,在拮抗劑(NPS-2390)組減少(P0.05)。結(jié)論:抑制鈣敏感受體的表達(dá)能夠有效減輕海馬神經(jīng)元的缺血再灌注損傷,抑制缺血再灌注后神經(jīng)元細(xì)胞凋亡,其機(jī)制與抑制ERK1/2的磷酸化有關(guān)。
[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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R743.3

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