Rho激酶对大鼠海马神经元骨架相关蛋白mRNA表达的影响  

Effects of Rho kinase on the cytoskeleton-related proteins mRNA in rat hippocampal neurons

作　　者：张忠其[1] 莫宏波[2] 李斌[1] 张文斌[3] 龚晓兵[4] 郭国庆[1] 

机构地区：[1]暨南大学医学院解剖学教研室,广东广州510632 [2]暨南大学医学院生物化学教研室,广东广州510632 [3]暨南大学第一附属医院外科,广东广州510630 [4]暨南大学第一附属医院消化内科人工肝室,广东广州510630

出　　处：《暨南大学学报（自然科学与医学版）》2010年第4期341-345,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广东省自然科学基金项目(8451063201000193);广东省医学科研基金项目(A2008344)

摘　　要：目的:探讨Rho激酶对大鼠海马神经元突起生长和骨架相关蛋白mRNA表达的影响。方法:用Rho激酶的抑制剂Y-27632和激动剂溶血磷脂酸(LPA)干预海马神经元,观察突起的生长并用RT-PCR检测大鼠海马神经元神经丝结合蛋白(Dbn1)、微丝肌动蛋白(F-actin)、微管相关蛋白(Tau)、α-微管蛋白(α-tubulin)mRNA的表达。结果:用LPA干预2 h后突起从远端逐渐退缩,长度缩短,细小突起退缩明显;Y-27632干预2 h后细胞胞体和突起的远侧端新生出细小突起。RT-PCR实际扩增长度与设计长度相吻合。内参照β-actin电泳条带在不同分组灰度较均一,呈高表达。Dbn1呈较高水平表达,激动剂LPA组表达下降(P<0.05),抑制剂Y-27632组表达量增多(P<0.05);F-actin和Tau表达呈低水平,激动剂作用后均使表达量相应的减少(P<0.05),抑制剂组表达增多(P<0.05);α-tubulin表达量最高,激动剂组表达量出现明显下降(P<0.05),抑制剂组表达增加(P<0.05)。结论:激活Rho激酶下调Dbn、F-actin、Tau、α-tubulin mRNA的表达并诱导突起缩短,抑制则增加其表达并促进突起生长。Aim：To investigate the effects of Rho kinase on the cytoskeleton-related proteins mRNA and neurites outgrowth in rat hippocampal neurons. Methods： Cultured rat hippocampal neurons were treated with Rho kinase activator LPA and inhibitor Y-27632,and neurites outgrowth was observed by using invert microscope.Expression of mRNA of Dbn1,F-actin,Tau,α-tubulin was detected by RT-PCR.Results： The neurites collapsed gradually from the distal of the neurons after treated with LPA for 2 hours,especially for the small neurites.However,small new neurites outgrew from the distal of the cell bodies and primitive neurites after incubation with Y-27632 for 2 hours.Results of RT-PCR,we found that the expression of β-actin was similar and at high levels in all groups.Expression of Dbn1 mRNA was at a relatively high level in all groups,but decreased in LPA group（P0.05） and increased in Y-27632 group（P0.05）;The expression of F-actin and Tau was similar to Dbn1 mRNA.Although F-actin and Tau were expressed at a lower levels in all groups,the expression of both genes declined after intervention with LPA（P0.05） and increased after intervention with Y-27632（P0.05）;The higher expression of α-tubulin mRNA was detected in neurons,meanwhile LPA significantly down-regulate its expression（P0.05）,and Y-27632 could significantly up-regulated its expression（P0.05）.Conclusion：Activating Rho kinase could down-regulate the expression levels of Dbn1,F-actin,Tau,α-tubulin mRNA and induce shortening of neurites.On the other hand,inhibiting Rho kinase could up-regulate the expression levels and promote neurites outgrowth.

关 键 词：RHO激酶 神经元 突起 生长 大鼠 海马 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] Q954.6[医药卫生—基础医学]