摘要
目的:探讨小续命汤对大鼠急性脑缺血再灌注后线粒体自噬中PINK1/Parkin通路及神经细胞凋亡的影响。方法:将120只大鼠随机分为假手术组、模型组、小续命汤组、小续命汤+线粒体自噬抑制剂组。采用线栓法制备右侧大脑中动脉栓塞(MCAO)大鼠模型,栓塞1 h后恢复灌注24 h,再灌注24 h后各组均选取20只。各组大鼠均于造模前连续灌胃给药5 d,术后24 h观察各组大鼠神经行为学评分、损伤部位海马区神经元病理变化、脑梗死体积,采用透射电镜观察脑损伤部位线粒体自噬情况,通过Western blotting法检测自噬相关蛋白P62、LC3以及PINK1、Parkin蛋白表达情况。结果:与模型组比较,小续命汤组大鼠海马区神经元及脑组织超微结构病变均明显减轻,Longa评分明显降低(P<0.05),大脑梗死比例明显缩小(P<0.05),P62表达水平明显下调(P<0.05),PINK1蛋白表达水平、Parkin蛋白表达水平、LC3-Ⅱ/LC3-Ⅰ比值明显升高(P<0.05);与小续命汤组比较,小续命汤+线粒体自噬抑制剂组大鼠海马区神经元及脑组织超微结构病变均明显加重,Longa评分明显升高(P<0.05),大脑梗死比例明显增大(P<0.05),P62表达水平明显上调(P<0.05),PINK1蛋白表达水平、Parkin蛋白表达水平、LC3-Ⅱ/LC3-Ⅰ比值明显降低(P<0.05)。结论:线粒体自噬可以缓解急性脑缺血再灌注后脑损伤程度,小续命汤可以通过PINK1/Parkin通路适度上调线粒体自噬,及时清除受损线粒体,减少大鼠神经细胞凋亡。
Objective: To investigate the effects of Xiaoxuming Decoction on PINK1/Parkin pathway and neuronal apoptosis in mitochondrial autophagy after acute cerebral ischemia-reperfusion in rats. Methods: A total of 120 rats were randomly divided into Sham operation group(Sham group), model group(I/R group), Xiaoxuming Decoction group(XXMD group), and Xiaoxuming Decoction + mitochondrial autophagy inhibitor group(XXMD+Mdivi-1 group). The rat model of right middle cerebral artery embolization(MCAO) was prepared by thread embolism method. The rats were perfused for 24 hours after embolization for 1 hour, and 20 rats were selected in each group after reperfusion for 24 hours. The rats in each group were continuously administered by gavage for 5 days before modeling. 24 hours after operation, the neurobehavioral score, pathological changes of neurons in hippocampus and cerebral infarction volume of rats in each group were observed. The mitochondrial autophagy in brain injury was observed by transmission electron microscope, and the expressions of autophagy related proteins P62, LC3, PINK1 and Parkin were detected by Western blotting. Results: Compared with the model group, the ultrastructural lesions of neurons and brain tissue in the hippocampus of rats in the XXMD group were significantly reduced, Longa score was significantly decreased(P<0.05), the proportion of cerebral infarction was significantly decreased(P<0.05), the expression level of P62 was significantly down-regulated(P<0.05), the protein expression levels of PINK1, Parkin and LC3-II/LC3-I were significantly increased(P <0.05). Compared with the XXMD group, the ultrastructural lesions of neurons and brain tissue in the hippocampus of rats in the XXMD+Mdivi-1 group were significantly aggravated, Longa score was significantly increased(P<0.05), the proportion of cerebral infarction was significantly increased(P<0.05), the expression level of P62 was significantly up-regulated(P<0.05),the protein expression levels of PINK1, Parkin and LC3-II/LC3-I were significantly decreased(P<0.05). Conclusion:Mitochondrial autophagy can alleviate the degree of brain injury after acute cerebral ischemia-reperfusion.Xiaoxuming Decoction can moderately up regulate mitochondrial autophagy through PINK1/Parkin pathway, timely remove damaged mitochondria, and reduce neuronal apoptosis in rats.
作者
王连辉
米国青
薛满中
WANG Lian-hui;MI Guo-qing;XUE Man-zhong(Tangshan Ninth Hospital,Tangshan Hebei 063000,China)
出处
《中医药导报》
2022年第1期36-40,44,共6页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
基金
河北省2018年度医学科学研究重点课题计划(201802623)。
