大鼠脑缺血再灌注后自噬的变化及灯盏生脉胶囊干预的影响  

Effects of Dengzhan Shengmai Capsules on Autophagy after Cerebral Ischemia-reperfusion in Rats

作　　者：陈兴泳[1] 汪银洲[1] 雷惠新[1] 张旭[1] 

机构地区：[1]福建医科大学省立临床医学院,福建省立医院神经内科,福建福州市350001

出　　处：《中国康复理论与实践》2015年第7期773-779,共7页Chinese Journal of Rehabilitation Theory and Practice

基　　金：福建省自然科学基金面上项目(No.2013J01275;No.2014J01401);福建省卫生厅中医药研究课题(No.wzzy201304)

摘　　要：目的观察大鼠脑缺血再灌注后自噬的变化及灯盏生脉胶囊干预的影响。方法 Sprague-Dawley大鼠50只分为假手术组、3-甲基腺嘌呤(3-MA)组、3-MA对照组、灯盏生脉胶囊(DZSM)组和DZSM对照组,每组10只。除假手术组外,其他大鼠线栓法制作大脑中动脉阻塞1 h再灌注模型。3-MA组和3-MA对照组造模前1 h立体定位侧脑室内注射3-MA或生理盐水,DZSM组和DZSM对照组从造模后4 h起每天予DZSM 20 mg/kg或生理盐水灌胃,共3 d。再灌注3 d后,各组采用Longa评分检测,免疫荧光染色和Western blotting检测脑组织微管相关蛋白1轻链3(LC3)、Beclin1表达,化学荧光法检测DZSM组和DZSM对照组活性氧水平。结果与假手术组相比,脑缺血再灌注后LC3、Beclin1表达显著增加(P<0.001),尤其在梗死灶周边的皮层和纹状体区。3-MA组LC3、Beclin1阳性细胞计数均显著少于3-MA对照组(均P<0.001),Longa评分显著低于3-MA对照组(P<0.001)。DZSM组LC3-Ⅱ、Beclin1蛋白表达显著低于DZSM对照组(均P<0.001),活性氧水平、Longa评分也显著低于DZSM对照组(P<0.001)。结论抑制局灶性脑缺血再灌注后自噬反应可保护神经功能。DZSM胶囊的作用机制包括抑制自噬反应。Objective To study the effects of Dengzhan Shengmai （DZSM） capsules on the autophagy in brains after cerebral Isch-emia-reperfusion in rats. Methods 50 Sprague-Dawley rats were divided into sham group （n=10）, 3-methyladenine （3-MA） group （n=10）, 3-MA-control group （n=10）, DZSM group （n=10） and DZSM-control group （n=10）. The middle cerebral arteries were occluded for 1 hour and re-perfused in all the rats except the sham group. The 3-MA and 3-MA-control groups were injected 3-MA or normal saline （NS） into the right lateral ventricle 1 hour before operation. The DZSM and DZSM-control groups accepted DZSM or NS by gavage daily for 3 days since 4 hours after reperfusion. The rats were assessed with Longa＆#39;s score 3 days after operation, and the expression of microtubule-associat-ed protein 1 light chain 3 （LC3） and Beclin1 in the brain were detected with immunofluorescent labeling and Western blotting;and the level of reactive oxygen species （ROS） were detected with chemiluminescence in the DZSM and DZSM-control groups. Results The expression of LC3 and Beclin1 increased in the ipsilateral ischemic hemisphere, especially in the cortex and striatum surrounding the infarct core （P〈0.001, compared with the sham group）. The expression of LC3 and Beclin1 decreased in the 3-MA group compared with the 3-MA-control group （P〈0.001）, with the decrease of Longa＆#39;s score （P〈0.001）. The expression of LC3-II and Beclin1 decreased in the DZSM group com-pared with the DZSM-control group （P〈0.001）, with the decrease of Longa＇s score （P〈0.001） and level of ROS （P〈0.001）. Conclusion Inhi-bition of autophagy after focal cerebral ischemia-reperfusion plays a role in neuroprotection, which may be a way of DZSM to work.

关 键 词：脑缺血再灌注损伤 自噬 灯盏生脉胶囊 大鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]