摘要
目的研究小鼠脑出血(ICH)后自噬是否被激活及NF-κB信号通路对ICH诱发的自噬相关蛋白表达的影响。方法正常昆明小鼠纹状体注射胶原酶Ⅳ建立脑出血模型,或于脑出血模型建立前5 min,侧脑室给药SN50(0.1μg/μl)或生理盐水1μl,随机分为脑出血组、SN50处理组和生理盐水处理组,同时设立空白对照组。每组分别于脑出血1 h,6 h,24 h,48 h,7 d后(5只/时间点)断头取脑,脑出血各组取出血区及周边脑组织进行免疫印记实验检测Beclin-1、Bcl-2、LC3蛋白表达水平或激活情况,SN50处理组和生理盐水处理组各时间点取出血区及周边脑组织进行免疫印记实验检测LC3蛋白激活情况。结果小鼠脑出血后出血区及出血周边脑组织Beclin-1蛋白表达上调、Bcl-2蛋白表达下调,以6～48 h时最为显著(P<0.05);Beclin-1/Bcl-2比值随时间变化逐渐增高,以6～48 h时最为显著(P<0.05);LC3被激活,随时间变化蛋白表达逐渐增加,48 h时达高峰,至7 d时仍较空白对照组有显著差异(P<0.05)。脑出血后,SN50处理组LC3蛋白激活高于生理盐水处理组,以6～48 h时最为显著(P<0.05)。结论脑出血后自噬被激活,这种激活的机制之一涉及到Beclin-1/Bcl-2比值的增加;NF-κB特异性抑制剂SN50促进了脑出血后自噬相关蛋白的上调表达。
Objective To investigate whether autophagy was activated and the effects of SN50,an inhibitor of NF-κB,on expression of autophagy-related proteins after intracerebral hemorrhage（ICH） in mice.Methods Intracerebral hemorrhage was produced by injection of collagenaseⅣinto striatum in adult Kun＇ ming mice,and mice were randomly divided into the SN50 pretreatment group,the saline vehicle group,the ICH group,and the control group.The SN50 and saline groups were produced by intracere-broventricular （i.c.v.） administration of SN50（0.1μg/μl,1μl） or saline（1μl） 5 min before ICH. Animals were sacrificed 1 h,6 h,24 h,48 h and 7 d after ICH in each group,and brains were harves- ted.Region of hemorrhage and perilesional were dissected for Western blot analysis of Beclin-1,Bcl-2 and LC3.Results Beclin-1 was significantly up-regulated and Bcl-2 significantly down-regulated from 6 to 48 h post ICH in mice（P〈0.05）.The ratio of Beclin-1/Bcl-2 was increased from 6 to 48 h post ICH （P〈0.05）.LC3 was significantly activated from 1 to 7 d after ICH in mice（P〈0.05）.Up-regulation of LC3 activation was enhances in SN50-pretreated groups compared with that in saline vehicle groups. Conclusion Autophagy is activated and ratio of Beclin-l/Bcl-2 is one of the regulation mechanisms in ICH mice.Inhibition of NF-κB signal pathway by SN50 enhances the up-regulation of autophagy-related protein induced by ICH in mice.
出处
《苏州大学学报（医学版）》
CAS
北大核心
2011年第1期30-34,共5页
Suzhou University Journal of Medical Science
基金
国家自然科学基金资助项目(30571909
0872666)
上海市法医学重点实验室开放基金资助项目(KF0904)
