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

doi:10.12122/j.issn.1673-4254.2026.07.13

Abstract

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

Kelin SHI, Xiaochuan QI, Shixiang XU, Yuanyuan GAO, Mingzhe FANG, Juncheng HU, Chunjing WANG, Changqing LIU, Yu GUO. CHIR99021 alleviates neuroinflammation in a mouse model of Parkinson's disease by inhibiting the GSK-3β/NF-κB pathway[J]. Journal of Southern Medical University, 2026, 46(7): 1593-1600.

Figures/Tables 5

Fig.1 Effect of CHIR99021 on motor function of MPTP-PD mice. A: Flow chart of MPTP-PD and MPTP+CHIR99021 modeling in the mice. B: Pole test turning time. C: Pole test down time. D: Rotarod test stick time. E: Rotarod test maximum rotation. F: Total distance traveled by mice in the open-field test. G-I: Movement trajectory of the mice. **P<0.01, ***P<0.001 (n=8).

Fig.2 CHIR99021 reduces the release of pro-inflammatory factors induced by MPP⁺. A, B: Western blotting for detecting expression levels of the inflammation-related markers. C: Cell immunofluorescence staining (Scale bar=10 μm). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Fig.3 CHIR99021 reduces the release of pro-inflammatory factors induced by MPTP. A, B: Western blotting for detecting expression levels of neuroinflammation-related markers in mouse substantia nigra tissue. C: Immunofluorescence staining of substantia nigra tissue (Scale bar=10 μm). *P<0.05, **P<0.01, ***P<0.001 .

Fig.4 Effect of CHIR99021 on expression level of autophagy markers in SH-SY5Y cells. A, B: Western blotting for detecting expression levels of autophagy markers in SH-SY5Y cells. C: Immunofluorescence staining of the cells (Scale bar=10 μm). *P<0.05, **P<0.01, **P<0.001.

Fig.5 CHIR99021 activates autophagy function in the mouse brain. A, B: Western blotting for detecting expression levels of neuroautophagy-related markers in mouse substantia nigra tissue. C: Immunofluorescence staining of the substantia nigra tissue (Scale bar=10 μm). *P<0.05, **P<0.01, ***P<0.001.

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