南方医科大学学报

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (7): 1593-1600.doi: 10.12122/j.issn.1673-4254.2026.07.13

• • 上一篇    

CHIR99021通过抑制GSK-3β/NF-κB信号通路减轻帕金森病小鼠的神经炎症

石科林1,2(), 齐霄川1,3, 徐世祥1,2, 高缘圆1,4, 房明哲1,4, 胡俊诚1,4, 王春景1,2, 刘长青1,2, 郭俣1,3()   

  1. 1.蚌埠医科大学，安徽省神经再生技术与医用新材料工程研究中心，安徽 蚌埠 233000
    2.蚌埠医科大学，生命科学学院，安徽 蚌埠 233000
    3.蚌埠医科大学，检验医学院，安徽 蚌埠 233000
    4.蚌埠医科大学，第一临床医学院，安徽 蚌埠 233000
  • 收稿日期:2025-11-11 出版日期:2026-07-20 发布日期:2026-07-20
  • 通讯作者: 郭俣 E-mail:shikelin00@163.com;guoyu@bbmu.edu.cn
  • 作者简介:石科林，在读硕士研究生，E-mail: shikelin00@163.com
  • 基金资助:
    国家自然科学基金(82371382);安徽省自然科学基金(2308085MH256);蚌埠医科大学龙湖英才项目(LH250102002);蚌埠医科大学自然科学研究项目资助(2024byzd024)

CHIR99021 alleviates neuroinflammation in a mouse model of Parkinson's disease by inhibiting the GSK-3β/NF-κB pathway

Kelin SHI1,2(), Xiaochuan QI1,3, Shixiang XU1,2, Yuanyuan GAO1,4, Mingzhe FANG1,4, Juncheng HU1,4, Chunjing WANG1,2, Changqing LIU1,2, Yu GUO1,3()   

  1. 1.Anhui Engineering Research Center for Neural Regeneration Technology and Medical New Materials, Bengbu Medical University, Bengbu 233000, China
    2.School of Life Sciences, Bengbu Medical University, Bengbu 233000, China
    3.School of Laboratory Medicine, Bengbu Medical University, Bengbu 233000, China
    4.First School of Clinical Medicine, Bengbu Medical University, Bengbu 233000, China
  • Received:2025-11-11 Online:2026-07-20 Published:2026-07-20
  • Contact: Yu GUO E-mail:shikelin00@163.com;guoyu@bbmu.edu.cn
  • Supported by:
    National Natural Science Foundation of China(82371382)

RichHTML

9

PDF

110

摘要：

目的 探讨CHIR99021抑制GSK-3β/NF-κB通路改善帕金森病神经炎症的作用。 方法 SH-SY5Y细胞加入1-甲基-4-苯基吡啶（MPP+）（1 mmol/L）培养24 h，构建MPP-PD细胞模型，治疗组SH-SY5Y细胞先经CHIR99021（3μmol/L）预处理2 h，再加入MPP+（1 mmol/L）培养24 h。将野生型（WT）小鼠连续5 d腹腔注射1-甲基-4-苯基-1，2，3，6-四氢吡啶（MPTP）构建MPTP-PD动物模型，治疗组连续7 d腹腔注射CHIR99021，从第3天开始间隔12 h再腹腔注射MPTP，连续注射5 d。以WT型小鼠作对照，通过旷场实验、爬杆实验、疲劳转棒实验评估各组运动行为能力。通过Western blotting 和免疫荧光检测CHIR99021在体内和体外抑制GSK-3β/NF-κB通路信号调控NLRP3炎症小体激活和细胞自噬改善帕金森病神经炎症的作用。 结果 与对照组相比，MPP⁺-PD细胞模型中，p-GSK-3β （Ser9）的蛋白表达水平下调（P<0.01），而总GSK-3β蛋白量差异没有统计学意义。p-NF-κB p65 （Ser536）的蛋白表达水平上调（P<0.01），而总NF-κB p65 （Ser536）蛋白量差异没有统计学意义。同时ASC、NLPR3、Caspase-1炎症小体表达上调（P<0.01）。而在MPP⁺+CHIR99021组中，p-GSK-3β、NF-κB及炎症小体蛋白表达下调.与对照组相比，MPTP-PD小鼠模型中，小鼠运动功能下降（P<0.001），脑内TH神经元减少（P<0.01）。黑质部位的p-GSK-3β （Ser9）水平同样降低（P<0.01），总GSK-3β蛋白量变化差异无统计学意义。而p-NF-κB p65 （Ser536）水平同样显著升高（P<0.01），总NF-κB p65蛋白量表达变化差异无统计学意义，ASC、NLPR3、Caspase-1炎症小体表达上调（P<0.01）。而MPTP-CHIR99021组较MPTP-PD组相比，p-GSK-3β、NF-κB及炎症小体蛋白表达下调（P<0.01）。 结论 CHIR99021通过抑制GSK-3β/NF-κB信号通路，显著改善帕金森病（PD）小鼠的运动功能，减轻神经炎症，并激活细胞自噬以维持细胞内环境稳态。靶向GSK-3β可同时调控PD多个关键病理环节，具有作为多功能治疗靶点的潜力。

关键词: 帕金森病, CHIR99021, GSK-3β, NF-κB, 神经炎症

Abstract:

Objective To investigate the effect of CHIR99021 for improving neuroinflammation in a mouse model of Parkinson's disease (PD) and the role of the GSK-3β/NF-κB pathway in mediating this effect. Methods In the cell experiment, induced SH-SY5Y cells were pretreated with 3 μmol/L CHIR99021 for 2 h prior to MPP+ treatment. In a male C57BL/6 mouse model of MPTP-induced PD, the effect of intraperitoneal injection of CHIR99021 for 7 days on motor function of the mice were assessed using behavioral tests. Western blotting and immunofluorescence staining were used to detect the changes in the GSK-3β/NF-κB pathway, NLRP3 inflammasome and autophagy in both SH-SY5Y cells and the mouse models. Results MPP+-treated SH-SY5Y cells showed significantly downregulated expression of p-GSK-3β protein and upregulated expressions of p-NF-κB p65, ASC, NLPR3, and caspase-1, and these changes were obviously attenuated by pretreatment with CHIR99021. Compared with normal mice, the PD mouse models showed significantly decreased motor function and reduced TH neurons in the brain with lowered expression of p-GSK-3β, increased expression of p-NF-κB p65 (Ser536), and upregulated ASC, NLPR3, and caspase-1 expressions. Treatment with CHIR99021 significantly increased the expression of p-GSK-3β, and reduced the expression of p-NF‑κB p65 (Ser536), ASC, NLPR3, and caspase-1. Conclusion CHIR99021 inhibits the GSK-3β/NF‑κB signaling pathway to improve motor function of PD mice, reduce neuroinflammation, and activate autophagy to maintain the homeostasis of intracellular environment, suggesting that targeting GSK-3β, a potential multifunctional therapeutic target, can simultaneously regulate multiple key pathological links in PD.

Key words: Parkinson's disease, CHIR99021, GSK-3β, NF-κB, neuroinflammation