RhoA蛋白参与凝血酶对胎鼠大脑皮层神经元的损伤
RhoA단백삼여응혈매대태서대뇌피층신경원적손상
RhoA is involved in thrombin-induced neuron injury in the cortex of fetal rats.
  • 万方数据
    中华神经医学杂志 중화신경의학잡지
    CHINESE JOURNAL OF NEUROMEDICINE
    2012年 6期 570-574 ,共5页

    唐海%王兴启%沈霞%赵辉%高丽%陈浩%崔桂云

    당해%왕흥계%침하%조휘%고려%진호%최계운

    脑出血%凝血酶%RhoA蛋白%皮层神经元 腦齣血%凝血酶%RhoA蛋白%皮層神經元
    뇌출혈%응혈매%RhoA단백%피층신경원
    Intracerebral hemorrhage%Thrombin%RhoA%Cortex neuron
    目的 探讨凝血酶诱导的胎鼠大脑皮层神经元损伤与RhoA蛋白活化表达的关系.方法 体外培养胎鼠大脑皮层神经元8d后,采用Western blotting检测不同浓度凝血酶(0、1、10、30和100 U/mL)作用3h后及30 U/mL凝血酶作用不同时间(0、0.5、1、3和6h)对皮层神经元RhoA蛋白活化表达的影响；RhoA蛋白活化抑制剂Exoenzyme C3预处理0.5 h及30 U/mL凝血酶作用皮层神经元3h后,行Western blotting检测RhoA蛋白活化表达的情况,并采用Hoechst33258核染色及CCK-8法观察Exoenzyme C3预处理和未处理时凝血酶对皮层神经元的损伤情况. 结果 在浓度实验中,30 U/mL凝血酶作用皮层神经元3h后能明显增加RhoA膜蛋白表达,与0U/mL组相比差异有统计学意义(P＜0.05)；而各浓度凝血酶对RhoA总蛋白表达无明显影响.在时程实验中,与其他时间点相比,30 U/mL凝血酶在3h时能明显增加RhoA膜蛋白表达,差异有统计学意义(P＜0.05),但对RhoA总蛋白表达也无明显影响.在Exoenzyme C3干预实验中,Exoenzyme C3预处理组与凝血酶组相比,RhoA膜蛋白表达明显降低,差异有统计学意义(P＜0.05)；Hoechst3258核染色显示,与凝血酶组相比,Exoenzyme C3预处理组中核浓染细胞数量明显降低,差异有统计学意义(P＜0.05)；CCK-8法检测细胞活性显示,凝血酶组细胞存活率明显低于Exoenzyme C3预处理组,差异有统计学意义(P＜0.05). 结论 凝血酶诱导的胎鼠大脑皮层神经元损伤与RhoA膜蛋白的高表达即RhoA蛋白活化表达有关.
    목적 탐토응혈매유도적태서대뇌피층신경원손상여RhoA단백활화표체적관계.방법 체외배양태서대뇌피층신경원8d후,채용Western blotting검측불동농도응혈매(0、1、10、30화100 U/mL)작용3h후급30 U/mL응혈매작용불동시간(0、0.5、1、3화6h)대피층신경원RhoA단백활화표체적영향；RhoA단백활화억제제Exoenzyme C3예처리0.5 h급30 U/mL응혈매작용피층신경원3h후,행Western blotting검측RhoA단백활화표체적정황,병채용Hoechst33258핵염색급CCK-8법관찰Exoenzyme C3예처리화미처리시응혈매대피층신경원적손상정황. 결과 재농도실험중,30 U/mL응혈매작용피층신경원3h후능명현증가RhoA막단백표체,여0U/mL조상비차이유통계학의의(P＜0.05)；이각농도응혈매대RhoA총단백표체무명현영향.재시정실험중,여기타시간점상비,30 U/mL응혈매재3h시능명현증가RhoA막단백표체,차이유통계학의의(P＜0.05),단대RhoA총단백표체야무명현영향.재Exoenzyme C3간예실험중,Exoenzyme C3예처리조여응혈매조상비,RhoA막단백표체명현강저,차이유통계학의의(P＜0.05)；Hoechst3258핵염색현시,여응혈매조상비,Exoenzyme C3예처리조중핵농염세포수량명현강저,차이유통계학의의(P＜0.05)；CCK-8법검측세포활성현시,응혈매조세포존활솔명현저우Exoenzyme C3예처리조,차이유통계학의의(P＜0.05). 결론 응혈매유도적태서대뇌피층신경원손상여RhoA막단백적고표체즉RhoA단백활화표체유관.
    Objective To investigate the relationship between the activation of RhoA and thrombin-induced neuron injury in the cortex of fetal rats. Methods The neurons from the fetal ratcortex were culturedin vitro for 8 d; and then,they were treated by thrombin at different concentrations (0,1,10,30 and 100 U/mL) for 3 h,and by 30 U/mL thrombm at different times (0,0.5,1,3 and 6 h);Western blotting was used to examine the effects of these different treatments on the activation of RhoA.The neurons were pretreated with Exoenzyme C3,the RhoA inhibitor,for 0.5 h,and incubated with 30 U/ml thrombin for 3 h; and then,Western blotting was employed to examine the activation of Rho A; besides that,the injuries of these neurons with the presence and absence of Exoenzyme C3 were observed by Hoechst33258 staining and CCK-8 assay.Results The activation of RhoA expressing in membrane with the treatment of 30 U/mL thrombin for 3 h was significantly increased as compared with that under the treatment of 0 U/mL (P＜0.05); and the total RhoA showed no significant changes with the treatment of all concentrations. The 3 h site with thrombin (30 U/mL) could more significantly induce R hoA expression as compared with other time sites (P＜0.05),and the total RhoA showed no significant changes under the treatment of all time sites. Pretreatrnent of neurons with Exoenzyme C3 could significantly down-regulate RhoA level as compared with those without pretreatment (P＜0.05); meanwhile,Hoechst33258 staining indicated that the number of brightly stained neurons in the Exoenzyme C3presence group was dramatically decreased as compared with that in the Exoenzyme C3 absence group (P＜0.05), and CCK-8 assay showed that the cell survival rate in Exoenzyme C3 absence group significantly decreased as compared with that in the Exoenzyme C3 presence group (P＜0.05).Conclusion RhoA in membrane is related to the thrombin-induced neuron injury in the cortex of fetal
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