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[电针对帕金森病小鼠碱性成纤维细胞生长因子/糖原合酶激酶-3β通路的调节作用]

[Effect of electroacupuncture on regulation of the basic fibroblast growth factor/glycogen synthase kinase-3β pathway in Parkinson's disease mice].

作者信息

Li Yuan-Yuan, Bao Xiao-Li, Wang Yao, Wang Yan-Chun

机构信息

College of Acupuncture and Orthopedics, Hubei University of Traditional Chinese Medicine, Wuhan 430060, China.

Clinical College of Traditional Chinese Medicine, Hubei University of Traditional Chinese Medicine, Wuhan 430060.

出版信息

Zhen Ci Yan Jiu. 2025 Feb 25;50(2):167-173. doi: 10.13702/j.1000-0607.20230972.

Abstract

OBJECTIVES

To observe the effect of electroacupuncture (EA) on basic fibroblast growth factor (BFGF)/glycogen synthase kinase-3β (GSK-3β) pathway in Parkinson's disease (PD) mice, so as to explore the potential mechanisms of EA in treating PD.

METHODS

Fifty C57/BL6 male mice were randomly divided into the control, model, EA, inhibitor and EA+inhibitor (combination) groups, with 10 mice in each group. The PD mouse model was established by intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine. Mice in the EA group received EA stimulation at "Fengfu" (GV16), "Taichong"(LR3) and "Zusanli"(ST36) for 30 min, once daily for 12 days. The inhibitor group received gavage of fibroblast growth factor receptor (FGFR) inhibitor AZD4547 (12.5 mg·kg·d) for 12 consecutive days. The open box experiment was used to observe the autonomous movement ability of mice. The expression level of tyrosine hydroxylase (TH) in substantia nigra was detected by immunohistochemistry. The apoptosis rate of substantia nigra cells was measured by TUNEL method. The relative protein expression levels of BFGF, phosphorylated phosphatidylinositol 3-kinase (p-PI3K), phosphorylated protein kinase B (p-AKT), p-GSK-3β, B-lymphomatoma-2 gene (Bcl-2), and Bcl associated X protein (Bax) in substantia nigra were detected by Western blot.

RESULTS

After modeling, compared with the control group, the total movement distance was shorten and the duration of rest was prolonged in the open box experiment (<0.01), the average absorbance of TH in the substantia nigra of the midbrain was decreased (<0.01), and the apoptosis rate of substantia nigra cells in the midbrain was increased (<0.01), the protein expression levels of BFGF, p-PI3K, p-AKT, p-GSK-3β and Bcl-2 were decreased (<0.01), while the expression level of Bax protein was increased (<0.01) in the model group. Compared with the model and the combination groups, the total movement distance was prolonged and the duration of rest was shorten (<0.01), the average absorbance of TH was increased (<0.05), and the apoptosis rate was decreased (<0.01, <0.05), the expression levels of BFGF, p-PI3K, p-AKT, p-GSK-3β and Bcl-2 were increased (<0.01, <0.05), while the expression level of Bax was decreased (<0.05) in the EA group.

CONCLUSIONS

EA at GV16, LR3 and ST36 can alleviate motor disorders, reduce cell apoptosis, protect dopaminergic neurons in mice with PD, which may be related to its effect in regulating the BFGF/GSK-3β pathway.

摘要

目的

观察电针(EA)对帕金森病(PD)小鼠碱性成纤维细胞生长因子(BFGF)/糖原合酶激酶-3β(GSK-3β)通路的影响,以探讨电针治疗PD的潜在机制。

方法

将50只C57/BL6雄性小鼠随机分为对照组、模型组、电针组、抑制剂组和电针+抑制剂(联合)组,每组10只。通过腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶建立PD小鼠模型。电针组小鼠于“风府”(GV16)、“太冲”(LR3)和“足三里”(ST36)接受电针刺激30分钟,每日1次,共12天。抑制剂组连续12天灌胃成纤维细胞生长因子受体(FGFR)抑制剂AZD4547(12.5mg·kg·d)。采用旷场实验观察小鼠自主运动能力。免疫组织化学法检测黑质中酪氨酸羟化酶(TH)的表达水平。TUNEL法检测黑质细胞凋亡率。采用蛋白质免疫印迹法检测黑质中BFGF、磷酸化磷脂酰肌醇3激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)、p-GSK-3β、B淋巴细胞瘤-2基因(Bcl-2)和Bcl相关X蛋白(Bax)的相对蛋白表达水平。

结果

造模后,与对照组相比,旷场实验中模型组小鼠总运动距离缩短、静止时间延长(<0.01),中脑黑质TH平均吸光度降低(<0.01),中脑黑质细胞凋亡率升高(<0.01),模型组BFGF、p-PI3K、p-AKT、p-GSK-3β和Bcl-2蛋白表达水平降低(<0.01),而Bax蛋白表达水平升高(<0.01)。与模型组和联合组相比,电针组小鼠总运动距离延长、静止时间缩短(<0.01),TH平均吸光度升高(<0.05),凋亡率降低(<0.01,<0.05),BFGF、p-PI3K、p-AKT、p-GSK-3β和Bcl-2表达水平升高(<0.01,<0.05),而Bax表达水平降低(<0.05)。

结论

针刺GV16、LR3和ST36可缓解PD小鼠的运动障碍,减少细胞凋亡,保护多巴胺能神经元,其机制可能与调节BFGF/GSK-3β通路有关。

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