大鼠海马注射溶血磷脂酸诱导Rb蛋白磷酸化和凋亡的研究  

Involvement of lysophosphatidic acid in phosphorylation of phospho-Rb and apoptosis in rat hippocampal neurons

作　　者：李慧[1] 张兆辉[1] 汪宁[1] 刘美真[1] 黄刚[1] 张珊珊[1] 何爱燕[1] 

机构地区：[1]武汉大学人民医院神经内科,430060

出　　处：《卒中与神经疾病》2011年第2期75-77,105,共4页Stroke and Nervous Diseases

摘　　要：目的观察大鼠海马注射溶血磷脂酸(LPA)后磷酸化Rb蛋白的表达及神经元凋亡的变化。方法将60只Wistar大鼠分为实验组(n=20)注射LPA、对照组(n=20)注射PBS和拮抗剂组(n=20)注射LPA+苏拉明,利用脑立体定位技术在大鼠双侧海马微量注射溶血磷脂酸溶剂,于注射后12、24、48和72h各不同时间点采用免疫荧光组织化学方法测定该区域神经细胞中ser795位点磷酸化Rb蛋白(p-Rb,ser795)的表达水平;TUNEL技术检测神经细胞凋亡情况。结果实验组注射LPA后12h海马CA3区神经细胞中p-Rb表达增加,注射LPA24h后表达到达高峰,且均高于拮抗剂组和对照组(P<0.05)。LPA注射后12h可检测到凋亡细胞,24h TUNEL阳性细胞数达高峰,且阳性细胞数均多于拮抗剂组和对照组(P<0.05)。LPA注射后的12h和24h可观察到p-Rb和TUNEL分别以镶嵌和重叠的方式共定位。结论 LPA在动物水平上诱导了Rb蛋白的磷酸化并导致了神经细胞早期的凋亡,可能间接或者不参与其晚期凋亡过程。Objective To investigate the effects of Lysophosphatidic acid （LPA） on hyperphosphorylation of phospho-Rb （ser 795） and apoptosis in hippocampal neurons. Methods 60 adult rats were assigned to 3 groups： LPA injection groups, LPA ＋ suramin injection groups and vehiculurn injection groups. StereOtaxic microinjection of LPA or vehiculum into hippocampus was performed. At interval time of 12,24,48 and 72 hours after injection, the expression of Phospho-Rb（ser 795） was observed by immunohistochemical assay, and neuron apoptosis was detected by TUNEL. Results Compared with control groups, the expression of phospho-Rb modestly increased in CA3 sector of hippocampus at 12 hours, dramatically increased at 24 hours after injection （P〈0. 05）. Many TUNEL postive cells were detected at 12 hours,reached a maximum at 24 hours after injection（P〈0. 05）. Most phospho-Rb appeared to be localized in an overlapping and mosaic pattern within TUNEL at 12h and 24h after injection staining cells. Conehtsions In vivo,LPA can effectively induce hyperphosphorylation of Rb and phospho-Rb may be involved in the eariy stage of neuronal apoptosis in rat hippocampual neurons, it may not he involved or in an indirect way in the late stage of apoptosis.

关 键 词：溶血磷脂酸 p-Rb蛋白 细胞凋亡 

分 类 号：R741[医药卫生—神经病学与精神病学]