摘要
目的探讨LC3/mTOR自噬通路调控黄芪甲苷在大鼠脑缺血再灌注损伤中的作用和意义。方法将50只成年雄性Wistar大鼠构建大鼠脑缺血再灌注损伤(MCAO)模型,随机分为对照组、模型组、雷帕霉素(RAPA)组、3-MA组、黄芪甲苷(AST)组。采用灌胃方法在造模前分别灌注雷帕霉素,3-MA和黄芪甲苷。按Longa评分标准进行神经功能缺陷评分;计算梗死灶体积和HE染色观察脑损伤情况;采用实时荧光定量PCR(RT-PCR)和蛋白免疫印迹法(Western blot)检测mTOR、LC3、Beclin-1、ATG5、SQSTM1/p62和Caspase-3mRNA和蛋白质表达。结果与模型组相比,RAPA组、3-MA组、AST组Longa评分显著降低,梗死面积(%)显著减少,差异有统计学意义(P<0.05)。黄芪甲苷治疗后,大鼠Longa评分最低,梗死面积(%)最少,差异有统计学意义(P<0.05)。与模型组相比,AST组mTOR、p62和Caspase-3mRNA和蛋白质水平显著降低,LC3和Beclin-1mRNA和蛋白质水平显著升高,差异有统计学意义(P<0.05)。结论黄芪甲苷可以有效治疗和缓解大鼠MCAO,LC3/mTOR自噬通路在此过程中发挥重要的作用。
Objective To investigate the role of LC3 / mTOR autophagy pathway in the regulation of astragaloside in cerebral ischemia-reperfusion injury in rats. Methods Fifty adult male Wistar rats were used to establish a model of middle cerebral artery occlusion （MCAO） in rats.The rats were randomly divided into control group,model group,rapamycin group,3-MA group,Astragaloside A （AST） group.The gavage method was used to perfuse rapamycin,3-MA and astragaloside respectively before modeling.Longa score was used to evaluate neurological deficit.Infarct volume and HE staining were used to evaluate brain injury.Real-time PCR and Western blot were used to detect mTOR,LC3,Beclin-1,ATG5,SQSTM1/p62 and Caspase-3 mRNA And protein expression. Results Compared with the model group,the Longa score of RAPA group,3-MA group and AST group decreased significantly and infarct area （%） decreased significantly （ P 〈0.05）.Astragaloside treatment,the Longa score the lowest,infarct size （%） least,the difference was statistically significant （ P 〈 0.05 ）.The mRNA and protein levels of mTOR,p62 and Caspase-3 in AST group were significantly lower than those in model group,while the contents of LC3 and Beclin-1 mRNA and protein were significantly increased in AST group （ P 〈0.05）. Conclusion Astragaloside can effectively treat and relieve cerebral ischemia-reperfusion injury in rats,and LC3/mTOR autophagy pathway plays an important role in this process.
作者
史楠
张燕
李晋峰
金磊
张蔚
赵梅珍
罗峥
赵玫
SHI Nan;ZHANG Yan;LI Jinfeng;JIN Lei;ZHANG Wei;ZHAO Meizhen;LUO Zheng;ZHAO Mei(Department of Neurology,Zhoupu Hospital Affiliated to Shanghai Health Medical College,Shanghai 201318,China)
出处
《检验医学与临床》
CAS
2018年第22期3363-3366,共4页
Laboratory Medicine and Clinic
基金
上海健康医学院种子基金项目(HMSF-17-22-037)
上海市浦东新区重点专病项目(PWZzb2017-11)
