摘要
目的利用网络药理学分析、分子对接分析及动物实验验证探讨天麻钩藤饮(TGY)对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的亚急性帕金森病(PD)模型小鼠的运动功能障碍及分子机制的影响。方法90只8周龄C57BL/6小鼠随机分为6组(n=15):对照组、模型组、模型+坏死性凋亡抑制剂NEC-1组(6.5 mg/kg)、模型+不同剂量的天麻钩藤饮治疗(TGY-L组2.5 g/kg;TGY-H组5 g/kg)、模型+阳性对照美多芭组(76 mg/kg)。采用连续5 d腹腔注射MPTP(30 mg/kg)构建亚急性PD小鼠模型。随机选取小鼠进行运动能力测试(n=6);Western blotting检测小鼠纹状体中酪氨酸羟化酶(TH)和α-突触核蛋白(α-syn)、坏死性凋亡相关蛋白受体相互作用蛋白1(RIPK1)、RIPK3和混合谱系激酶域样(MLKL)及磷酸化的表达情况(n=4),并基于网络药理学和分子对接分析方法对天麻钩藤饮调控坏死性凋亡治疗PD的关键靶点进行预测。结果与对照组相比,PD组小鼠出现运动功能障碍,α-syn表达增加(P<0.001),TH蛋白表达下降(P<0.001)。连续6 d的NEC-1治疗后运动功能得到改善,p-RIPK1/RIPK1、p-RIPK3/RIPK3、p-MLKL/MLKL和α-syn表达减少(P<0.05或P<0.001),以及TH表达增加(P<0.01)。网络药理学预测结果显示天麻钩藤饮可调控坏死性凋亡通路中的RIPK1来改善神经退行性疾病,且分子对接结果表明天麻钩藤饮中的化合物与RIPK1存在相互作用。动物实验结果证实,低、高剂量的天麻钩藤饮改善了PD小鼠的运动功能障碍,增加TH表达(P<0.01),降低α-syn和坏死性凋亡通路相关蛋白表达水平(P<0.05或P<0.01)。结论天麻钩藤饮通过抑制坏死性凋亡通路,促进TH蛋白表达和减轻α-syn蓄积,进而改善PD小鼠的运动功能障碍。
Objective To investigate the effects of formulated granules of Tianma Gouteng Yin(TGY)on motor deficits in a mouse model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced subacute Parkinson's disease(PD)and explore the possible molecular mechanisms.Methods Ninety C57BL/6 mice were randomized equally into 6 groups,including a control group,a PD model group,a NEC-1(6.5 mg/kg)treatment group,two TGY treatment groups at 5 and 2.5 g/kg,and a Madopar(76 mg/kg)treatment(positive control)group.Mouse models of PD were established by intraperitoneal injection of MPTP(30 mg/kg)for 5 consecutive days with the corresponding treatments for 15 days.The mice were randomly selected for motor function tests.Western blotting was used to detect the changes in expressions of TH,α-syn,RIPK1,RIPK3 and MLKL in the striatum of the mice.Network pharmacology analysis and molecular docking studies were performed to explore TGYmediated regulation of the necroptosis pathway for PD treatment.Results Compared with those in the control group,the PD model mice exhibited obvious motor deficits with significantly increasedα-syn protein expression and lowered TH protein expression in the striatum.Treatment with NEC-1 obviously improved motor deficits,inhibited the necroptosis pathway,and alleviated the changes in TH andα‑syn proteins in PD mice.Network pharmacology and molecular docking analyses suggested that the therapeutic effect of TGY in PD was associated with the modulation of RIPK1,a key protein in the necroptosis pathway.In PD mouse models,TGY treatment at the two doses significantly improved motor deficits of the mice,increased TH expression,and decreased the expressions ofα-syn and necroptosis-related proteins in the striatum.Conclusion TGY can effectively inhibit the necroptosis pathway,increase TH expression and decreaseα-syn expression in the striatum to improve motor deficits in PD mice.
作者
陈丹丹
任乾千
吕梦林
张宝文
刘醒然
张蒙
王阳
寇现娟
CHEN Dandan;REN Qianqian;LÜMenglin;ZHANG Baowen;LIU Xingran;ZHANG Meng;WANG Yang;KOU Xianjuan(School of Sports Medicine,Wuhan Sports University,Wuhan 430079,China;Hubei Provincial Key Laboratory of Exercise Training and Monitoring,Wuhan Sports University,Wuhan 430079,China;School of Physical Education,Guangxi University of Science and Technology,Liuzhou 545000,China;School of Physical Education and Health,Guangxi Medical University,Nanning 530021,China)
出处
《南方医科大学学报》
北大核心
2025年第8期1571-1580,共10页
Journal of Southern Medical University
基金
国家自然科学基金(81601228)
湖北省自然科学基金重点项目(2024AFD242)
湖北省高等学校优秀中青年科技创新团队计划项目(T2024019)
广西自然科学基金(2025GXNSFBA069048)
广西高校中青年教师科研基础能力提升项目(2023KY0365,2023KY0357)。
关键词
帕金森病
天麻钩藤饮
运动功能障碍
坏死性凋亡
Parkinson's disease
Tianma Gouteng Yin
motor deficits
necroptosis pathway
