摘要
目的 基于PTEN诱导激酶1(PTEN-induced putative kinase 1,PINK1)/帕金蛋白(Parkin)通路研究槲皮素减轻血管性痴呆(vascular dementia,VD)大鼠海马神经元损伤的作用机制。方法 通过双侧颈总动脉永久性结扎法建立VD大鼠模型,分为模型组及槲皮素低、中、高(25、50、100 mg/kg)组和欧来宁(14.8 g/kg)组,另取12只大鼠作为假手术组,给予药物干预21 d后,采用穿梭箱实验评价大鼠学习记忆能力,比较各组大鼠步入潜伏期(step-through latency,STL)及在暗室中花费的总时间(total time spent in the dark chamber,TDC);采用Nissl染色检测大鼠海马组织神经元损伤情况;采用透射电镜观察大鼠海马区神经元超微结构;采用ELISA法检测大鼠脑组织中乙酰胆碱(acetylcholine,Ach)、5-羟色胺(5-hydroxytryptamine,5-HT)水平和血清中白细胞介素-6(interleukin-6,IL-6)、IL-17、IL-1β水平;采用Western blotting检测大鼠脑组织中自噬相关蛋白与PINK1/Parkin信号通路相关蛋白表达。结果 与假手术组比较,模型组大鼠海马组织神经元损伤及超微结构受损严重,神经元内出现少量自噬体;STL显著降低(P<0.05),TDC显著升高(P<0.05);脑组织中Ach和5-HT水平显著降低(P<0.05),血清中IL-6、IL-17和IL-1β水平显著升高(P<0.05);脑组织中微管相关蛋白1轻链3II(microtubule-associated protein 1 light 3 II,LC3II)/LC3I、Beclin1、PINK1和Parkin蛋白表达水平均显著升高(P<0.05)。与模型组比较,槲皮素组大鼠海马组织神经元损伤及超微结构受损减轻,且神经元自噬现象增强;STL显著升高(P<0.05),TDC显著降低(P<0.05);脑组织中Ach及5-HT水平显著升高(P<0.05),血清中IL-6、IL-17和IL-1β水平均显著降低(P<0.05);脑组织中LC3II/LC3I、Beclin1、PINK1和Parkin蛋白表达水平均显著升高(P<0.05)。结论 槲皮素可激活PINK1/Parkin信号通路,降低VD大鼠炎性细胞因子水平,减轻海马神经元炎症损伤,提高大鼠学习记忆能力,改善其痴呆症状。
Objective To explore the mechanism of quercetin on alleviating hippocampal neuron damage in vascular dementia(VD)rats based on PTEN-induced putative kinase 1(PINK1)/Parkin signaling pathway. Methods VD rat model was established by permanent ligation of bilateral common carotid arteries,and were divided into model group,quercetin low-,medium-and high-dose(25,50,100 mg/kg) groups,and oxiracetam(14.8 g/kg) groups,and another 12 rats were selected as sham-operated group. After 21 d of drug intervention,shuttle box experiment was used to evaluate the learning and memory ability of rats,step-through latency(STL)and total time spent in the dark chamber(TDC) of rats in each group were compared;Nissl staining was used to detect neuronal damage in hippocampus of rats;Transmission electron microscope was used to detect the ultrastructural observation of neurons in rat hippocampus;ELISA was used to detect acetylcholine(Ach) and 5-hydroxytryptamine(5-HT) levels in brain tissue,and interleukin-6(IL-6),IL-17,and IL-1β levels in serum;Western blotting was used to detect expressions of autophagy-related proteins and PINK1/Parkin signaling pathway related proteins in brain tissues of rats in each group. Results Compared with sham operation group,neurons and ultrastructure of hippocampus in model group were seriously damaged,a small number of autophagosomes appear in neurons;STL was significantly decreased(P < 0.05),and TDC was significantly increased(P < 0.05);Levels of Ach and 5-HT in brain tissue were significantly decreased(P < 0.05),levels of IL-6,IL-17 and IL-1β in serum were significantly increased(P < 0.05);Microtubule-associated protein 1 light 3 II(LC3II)/LC3I,Beclin1,PINK1 and Parkin protein expression levels in brain tissue were significantly increased(P < 0.05). Compared with model group,neuronal damage and ultrastructural damage in hippocampus of rats in quercetin group were alleviated,and autophagy of neurons was enhanced;STL was significantly increased(P < 0.05),and TDC was significantly decreased(P < 0.05);Levels
作者
唐森
方建
高立功
黄一苇
TANG Sen;FANG Jian;GAO Li-gong;HUANG Yi-wei(Department of Neurology,Zhumadian Central Hospital,Zhumadian 463400,China;Department of Neurology,the First Affiliated Hospital of Henan University,Kaifeng 475001,China)
出处
《中草药》
CAS
CSCD
北大核心
2022年第20期6529-6535,共7页
Chinese Traditional and Herbal Drugs
基金
河南省医学科技攻关计划联合共建项目(LHGJ20210555)。
