天麻钩藤饮通过抑制坏死性凋亡通路改善帕金森病小鼠的运动功能障碍

doi:10.12122/j.issn.1673-4254.2025.08.01

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (8): 1571-1580.doi: 10.12122/j.issn.1673-4254.2025.08.01

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天麻钩藤饮通过抑制坏死性凋亡通路改善帕金森病小鼠的运动功能障碍

陈丹丹¹^,³(), 任乾千¹, 吕梦林¹, 张宝文¹, 刘醒然¹^,⁴, 张蒙¹, 王阳³, 寇现娟¹^,²()

^1.武汉体育学院，运动医学院，湖北武汉 430079
^2.武汉体育学院，运动训练监控湖北省重点实验室，湖北武汉 430079
^3.广西科技大学体育学院，广西柳州 545000
^4.广西医科大学体育与健康学院，广西南宁 530021

收稿日期:2025-03-27 出版日期:2025-08-20 发布日期:2025-09-05
通讯作者: 寇现娟 E-mail:1506087776@qq.com;kouxianjuan@126.com
作者简介:陈丹丹，在读博士研究生，E-mail: 1506087776@qq.com
基金资助:
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);广西高校中青年教师科研基础能力提升项目(2023KY0357)

Tianma Gouteng Granule improves motor deficits in mouse models of Parkinson's disease by regulating the necroptosis pathway

Dandan CHEN¹^,³(), Qianqian REN¹, Menglin LÜ¹, Baowen ZHANG¹, Xingran LIU¹^,⁴, Meng ZHANG¹, Yang WANG³, Xianjuan KOU¹^,²()

^1.School of Sports Medicine, Wuhan Sports University, Wuhan 430079, China
^2.Hubei Provincial Key Laboratory of Exercise Training and Monitoring, Wuhan Sports University, Wuhan 430079, China
^3.School of Physical Education, Guangxi University of Science and Technology, Liuzhou 545000, China
^4.School of Physical Education and Health, Guangxi Medical University, Nanning 530021, China

Received:2025-03-27 Online:2025-08-20 Published:2025-09-05
Contact: Xianjuan KOU E-mail:1506087776@qq.com;kouxianjuan@126.com
Supported by:
National Natural Science Foundation of China(81601228)

摘要/Abstract

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。

关键词: 帕金森病, 天麻钩藤饮, 运动功能障碍, 坏死性凋亡

Abstract:

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 TGY-mediated 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 increased α-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 and α‑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 of α-syn and necroptosis-related proteins in the striatum. Conclusion TGY can effectively inhibit the necroptosis pathway, increase TH expression and decrease α-syn expression in the striatum to improve motor deficits in PD mice.

Key words: Parkinson's disease, Tianma Gouteng Yin, motor deficits, necroptosis pathway

陈丹丹, 任乾千, 吕梦林, 张宝文, 刘醒然, 张蒙, 王阳, 寇现娟. 天麻钩藤饮通过抑制坏死性凋亡通路改善帕金森病小鼠的运动功能障碍[J]. 南方医科大学学报, 2025, 45(8): 1571-1580.

Dandan CHEN, Qianqian REN, Menglin LÜ, Baowen ZHANG, Xingran LIU, Meng ZHANG, Yang WANG, Xianjuan KOU. Tianma Gouteng Granule improves motor deficits in mouse models of Parkinson's disease by regulating the necroptosis pathway[J]. Journal of Southern Medical University, 2025, 45(8): 1571-1580.

图/表 5

图 1 PD模型成功

Fig. 1 Establishment of mouse models of MPTP/p-induced Parkinson's disease (PD). A: Schematic diagram of mice tested on the runway. B: Gait pattern and stride length in the control group. C: Gait pattern and stride length in the PD model group. D: Quantification of step cycle. E: Quantification of stride length. F: Expression of TH protein and a-syn protein in the striatum of the mice. Data are presented as Mean±SD if not specified otherwise and analyzed using one-way ANOVA. ##P<0.01,###P<0.001 vs Control. LF: Left front paw. RF: Right front paw. LH: Left hind. RH: Right hind.

图2 天麻钩藤饮改善PD小鼠的运动功能障碍

Fig.2 Tianma Gouteng Yin (TGY) at low (TGY-L) and high doses (TGY-H) improves motor deficits in PD mice by. A: Quantification of step cycle. B: Quantification of stride length. C: Quantification of latency to fall in rotarod test. D: Quantification of T-total in pole test. E: Quantification of T-turn in pole test. *P<0.05, **P<0.01, ***P<0.001 vs PD; ^P<0.01, ^^P<0.01, ^^^P<0.01 vs PD+TDY-L.

图3 网络药理学预测结果

Fig.3 Prediction results of network pharmacology. A: TGY and necroptosis intersection genes. B: PPI network diagram of the core target proteins. C: Bubble map of KEGG analysis of the key genes. D: Molecular docking result patterns of RIPK1 and compounds in TGY.

图4 NEC-1减轻PD小鼠运动功能障碍和病理蛋白蓄积

Fig.4 NEC-1 attenuates motor deficits and pathological protein accumulation in PD mice. A: Quantification of latency to fall in rotarod test. B: Quantification of T-total in pole test. C: Quantification of T-turn in pole test. D: Expression of key proteins of the necroptosis pathway in the striatum of the mice. E: Expression of TH and a-syn protein in the striatum of the mice. #P<0.05, ###P<0.001 vs Control; *P<0.05, **P<0.01, ***P<0.001 vs PD.

图5 天麻钩藤饮下调坏死性凋亡通路和病理蛋白表达

Fig.5 TGY attenuates the necroptosis pathway and pathological protein accumulation in PD mice. A: Expression of the key proteins of the necroptosis pathway in the striatum of the mice. B: Expression of TH protein and a-syn protein in the striatum of the mice.*P<0.05, **P<0.01, ***P<0.001 vs PD.

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天麻钩藤饮通过抑制坏死性凋亡通路改善帕金森病小鼠的运动功能障碍

Tianma Gouteng Granule improves motor deficits in mouse models of Parkinson's disease by regulating the necroptosis pathway

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