摘要
目的探索SIRT1-TLR4信号通路在1-甲基-4-苯基-1,2,3,6-四氢吡啶盐酸盐(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)帕金森病(PD)小鼠模型炎症反应中的作用。方法选用MPTP处理C57BL/6小鼠构建帕金森病小鼠模型,将小鼠随机分为对照组(Con组)、模型组(MPTP组)、激活剂组(RV+MPTP组)、抑制剂组(EX527+RV+MPTP组)。腹腔注射MPTP建立帕金森病小鼠模型,并给予白藜芦醇(RV)灌胃、SIRT1特异性抑制剂EX527腹腔注射处理,通过爬杆实验、旷场实验评估小鼠行为学改变,利用免疫荧光、Western blot等检测酪氨酸羟化酶(TH)、SIRT1、TLR4、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)等相关蛋白表达。结果和Con组相比,MPTP组小鼠爬杆总时间和爬杆转向时间明显增加,行走总路程、进入中心区域总距离、进入中心区域次数均明显减少(P<0.05);和MPTP组相比,RV+MPTP组小鼠爬杆总时间和爬杆转向时间明显缩短,行走总路程、进入中心区域总距离、进入中心区域次数均明显增加(P<0.05);和RV+MPTP组相比,EX527+RV+MPTP组小鼠爬杆总时间和爬杆转向时间明显增加,行走总路程、进入中心区域总距离、进入中心区域次数均明显减少(P<0.05)。免疫荧光检测发现,和Con组相比,MPTP组小鼠脑黑质区TH阳性神经元和SIRT1蛋白表达均明显减少,TLR4表达明显增加(P<0.05);和MPTP组相比,RV+MPTP组小鼠脑黑质区TH阳性神经元和SIRT1蛋白表达均明显增加,TLR4表达明显减少(P<0.05);和RV+MPTP组相比,EX527+RV+MPTP组小鼠脑黑质区TH阳性神经元和SIRT1蛋白表达均明显降低,TLR4表达明显增加(P<0.05)。蛋白质印迹检测发现,和Con组相比,MPTP组小鼠脑黑质中TH、SIRT1蛋白表达明显降低,TLR4、IL-6、TNF-α蛋白表达明显升高(P<0.05);和MPTP组相比,RV+MPTP组小鼠脑黑质中TH、SIRT1蛋白表达明显升高,TLR4、IL-6、TNF-α蛋白表达明显降低(P<0.05);和RV+MPTP组相比,EX527+RV+MPTP组小鼠小鼠脑黑质中TH、S
Objective To invstigate the role of SIRTI-TLR4 signaling pathway in the inflammatory response in a mouse model of MPTP Parkinson's disease(PD).Methods MPTP-treated C57BL/6 mice were selected to construct a mouse model of Parkinson's disease,and randomly divided into control group(Con group),model group(MPTP group),activator group(RV+MPTP group)and inhibitor group(EX527+RV+MPTP group).The mouse model of Parkinson's disease was established by intraperitoneal injection of MPTP and treated with resveratrol(RV)gavage and SIRTI-specific inhibitor EX527.Behavioral changes were assessed by pole climbing and open field experiments,and the expression of tyrosine hydroxylase(TH),SIRT1,TLR4,tumor necrosis factorα(TNF-α),interleukin 6(IL-6)and other related proteins were assessed by immunofluorescence and Western blot.Results Compared with the Con group,the mice in the MPTP group had significantly increased total pole climbing time and pole climbing turning time,and significantly reduced total walking distance,total distance into the central area,and number of times into the central area(P<0.05).Compared with the MPTP group,the mice in the RV+MPTP group had significantly shorter total pole climbing time and pole climbing and turning time,and significantly greater total walking distance,total distance into the central area,and number of times into the central area(P<0.05).Compared with the RV+MPTP group,mice in the EX527+RV+MPTP group had significantly increased total pole climbing time and pole climbing turning time,and significantly decreased total walking distance,total distance into the central area,and number of times into the central area(P<0.05).Immunofluorescence detection revealed that compared with the Con group,the expression of TH-positive neurons and SIRT1 protein in the nigrostriatal area of the brain was significantly reduced and TLR4 expression was significantly increased in the MPTP group(P<0.05).Compared with the MPTP group,the expression of TH-positive neurons and SIRTI protein was significantly increased
作者
邢瑶平
郭彦杰
郭清华
郑婷华
邢红霞
XING Yao-ping;GUO Yan-jie;GUO Qing-hua;ZHENG Ting-hua;XING Hong-xia(Departmentof Neurology,Ward 1,Xinxiang Medical University,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453000,Henan,China)
出处
《广东医学》
CAS
2023年第5期550-557,共8页
Guangdong Medical Journal
基金
河南省医学科技攻关计划联合共建项目(LHGJ20210548)
