《中国康复理论与实践》

《中国康复理论与实践》 ›› 2015, Vol. 21 ›› Issue (07): 773-779.

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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

陈兴泳，汪银洲，雷惠新，张旭   

  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2015-07-25 发布日期:2015-07-25

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

CHEN Xing-yong, WANG Yin-zhou, LEI Hui-xin, ZHANG Xu   

  1. Department of Neurology, Fujian Provincial Hospital, Fujian Medical University Shengli Clinical College, Fuzhou, Fujian 350001, China
  • Received:1900-01-01 Revised:1900-01-01 Published:2015-07-25 Online:2015-07-25

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被引次数

15

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。

关键词: 脑缺血再灌注损伤, 自噬, 灯盏生脉胶囊, 大鼠

Abstract: Objective To study the effects of Dengzhan Shengmai (DZSM) capsules on the autophagy in brains after cerebral Ischemia-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's score 3 days after operation, and the expression of microtubule-associated 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's score (P<0.001). The expression of LC3-II and Beclin1 decreased in the DZSM group compared 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 Inhibition of autophagy after focal cerebral ischemia-reperfusion plays a role in neuroprotection, which may be a way of DZSM to work.

Key words: cerebral ischemia-reperfusion injury, autophagy, Dengzhan Shengmai Capsules, rats