益气活血化浊解毒方对脑缺血再灌注损伤模型大鼠自噬相关蛋白Beclin-1、LC3-Ⅱ、mTOR表达的影响  

Effects of Yiqi Huoxue Huazhuo Jiedu Decoction on the expression of autophagy related proteins in brain tissue of rats with cerebral ischemia reperfusion injury

作　　者：崔怡娴 孙亚萍 石瑞 谢占维 孙玲玲 田军彪[1] CUI Yixian;SUN Yaping;SHI Rui(Hebei college of Chinese Medicine,Shijiazhuang,Hebei 050091,China;Graduate School of Hebei University of Traditional Chinese Medicine,Shijiazhuang,Hebei 050091,China)

机构地区：[1]河北省中医院/河北中医药大学第一附属医院,河北石家庄050011 [2]河北中医药大学研究生学院,河北石家庄050091

出　　处：《四川中医》2025年第2期98-104,共7页Journal of Sichuan of Traditional Chinese Medicine

基　　金：河北省科技厅重点研发计划项目(编号:20377710D);河北省中医药管理局科研计划重点项目(编号:Z2022008);河北中医学院科研能力提升重点项目(编号:KTZ2019028);河北中医学院科研能力提升一般项目(编号:KTZ2019013);河北中医学院研究生创新资助项目(编号:XCXZZSS2023025)。

摘　　要：目的:观察益气活血化浊解毒方对脑缺血再灌注损伤模型大鼠自噬相关蛋白Beclin-1、LC3-Ⅱ、mTOR表达的影响,初步探讨其作用机制。方法:将60只SPF级SD大鼠随机分为假手术组、模型组、尼莫地平组、中药低剂量组、中药中剂量组、中药高剂量组,每组10只。除假手术组外,其余各组按Zea-Longa线栓法制备大脑中动脉闭塞(MCAO)大鼠模型,造模成功后,各组大鼠分别予相应药物干预,给药3天后进行大鼠神经功能缺损评分;盐酸2,3,5-三苯基四氮(TTC)染色观察脑梗死范围;Nissl染色观察脑组织病理改变;Western blot检测大鼠脑组织mTOR、Beclin-1、LC3-Ⅱ蛋白相对表达量;qRT-PCR检测大鼠脑组织p-mTOR、Beclin-1、LC3-ⅡmRNA表达水平。结果:(1)与假手术组相比,模型组大鼠神经功能缺损评分增加(P<0.05);脑梗死体积百分比增大(P<0.05);细胞病理形态改变严重;Western blot检测结果显示Beclin-1、LC3-Ⅱ蛋白表达水平增多(P<0.05),mTOR蛋白表达水平降低(P<0.05)。qRT-PCR检测结果显示Beclin-1、LC3-ⅡmRNA表达水平均升高(P<0.05),p-mTOR mRNA表达水平降低(P<0.05)。(2)与模型组相比,各治疗组神经功能缺损评分均不同程度下降(P<0.05);脑梗死体积百分比均减少(P<0.05);细胞病理形态呈不同程度的改善,以中药高剂量组与尼莫地平组改善最为明显;Western blot检测结果显示Beclin-1、LC3-Ⅱ蛋白表达水平均降低(P<0.05),mTOR蛋白表达量均升高(P<0.05);qRT-PCR检测结果显示Beclin-1、LC3-ⅡmRNA表达水平均降低(P<0.05),p-mTOR mRNA表达水平均升高(P<0.05)。结论:益气活血化浊解毒方对脑缺血再灌注损伤大鼠缺血脑组织有一定改善作用,可能通过调节Beclin-1、LC3-Ⅱ、mTOR表达,抑制细胞自噬来实现。Objective:To Observe the protective effect of the Yiqi Huoxue Huazhuo Jiedu formula(YHJF)on brain tissue of rats with middle cerebral artery occlusion model,and exploring the effect of YHJF on autophagy and proteins of Beclin-1,LC3-Ⅱ,mTOR.Methods:Sixty SPF level SD rats were divided into the Sham group,the Model group,the Nimodipine group,the TCM-L group,the TCM-M group,and the TCM-H group,with 10 rats in each group.Except for the Sham group,all other groups prepared models of MCAO.Each group receives corresponding intervention measures.The mNSS method deficits in rats after 3 days.Observation of cerebral infarction area using TTC staining.Observation of pathological changes in brain tissue using Nissl staining.Western blot detection of relative expression levels of mTOR,Beclin-1,and LC3-Ⅱproteins.RT-PCR detection of p-mTOR,Beclin-1,and LC3-ⅡmRNA expression levels.Results:Compared with the sham group,the mNSS of the model group rats increased(P<0.05);the percentage of cerebral infarction volume increased(P<0.05);severe changes in cellular pathological morphology;the Western blot results showed an increase in the expression levels of Beclin-1and LC3-Ⅱproteins(P<0.05),while the expression levels of mTOR proteins decreased(P<0.05).The PCR detection results showed that the expression levels of Beclin-1and LC3-ⅡmRNA increased(P<0.05),while the expression levels of p-mTOR mRNA decreased(P<0.05).Compared with the model group,the mNSS scores in each treatment group decreased to varying degrees(P<0.05);the percentage of cerebral infarction volume decreased(P<0.05);the pathological morphology of cells showed varying degrees of improvement.The Western blot results showed that the expression levels of Beclin-1 and LC3-Ⅱproteins decreased(P<0.05),while the expression level of mTOR protein increased(P<0.05);the qRT-PCR detection results showed that the expression levels of Beclin-1 and LC3-ⅡmRNA decreased(P<0.05),while the expression levels of p-mTOR mRNA increased(P<0.05).Conclusion The YHJF has a certain improv

关 键 词：益气活血化浊解毒方 脑缺血再灌注损伤 自噬 BECLIN-1 LC3-Ⅱ MTOR 

分 类 号：R285[医药卫生—中药学]