40 Hz光闪烁刺激通过抑制STING信号通路改善帕金森病小鼠的认知功能障碍

doi:10.12122/j.issn.1673-4254.2026.10.04

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (10): 2325-2337.doi: 10.12122/j.issn.1673-4254.2026.10.04

• • 上一篇

40 Hz光闪烁刺激通过抑制STING信号通路改善帕金森病小鼠的认知功能障碍

任乾千¹(), 张宝文¹, 刘醒然¹^,², 丁伟利³^,⁴, 陈丹丹¹^,⁵, 寇现娟¹()

^1.武汉体育学院运动医学院，湖北武汉 430079
^2.广西医科大学体育与健康学院，广西南宁 530021
^3.燕山大学电气工程学院，河北秦皇岛 066004
^4.河北省智能康复及神经调控重点实验室，河北秦皇岛 066004
^5.广西科技大学体育学院，广西柳州 545006

收稿日期:2025-03-16 出版日期:2026-10-20 发布日期:2026-10-26
通讯作者: 寇现娟 E-mail:906371632@qq.com;kouxianjuan@126.com
作者简介:任乾千，在读硕士研究生，E-mail: 906371632@qq.com
基金资助:
湖北省高等学校优秀中青年科技创新团队(T2024019);教育部人文社会科学研究规划基金项目(25YJA890017)

40 Hz flicker light stimulation suppresses STING signaling to improve cognitive dysfunction in mice with Parkinson disease

Qianqian REN¹(), Baowen ZHANG¹, Xingran LIU¹^,², Weili DING³^,⁴, Dandan CHEN¹^,⁵, Xianjuan KOU¹()

^1.School of Sports Medicine, Wuhan Sports University, Wuhan 430079, China
^2.School of Sports and Health, Guangxi Medical University, Nanning 530021, China
^3.School of Electrical Engineering, Yanshan University, Qinhuangdao 066004, China
^4.Hebei Key Laboratory of Intelligent Rehabilitation and Neuromodulation, Qinhuangdao 066004, China
^5.School of Physical Education, Guangxi University of Science and Technology, Liuzhou 545006, China

Received:2025-03-16 Online:2026-10-20 Published:2026-10-26
Contact: Xianjuan KOU E-mail:906371632@qq.com;kouxianjuan@126.com

摘要/Abstract

摘要：

目的探究40 Hz光闪烁刺激对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）小鼠认知功能障碍的影响及其分子机制。方法将40只SPF级8周龄雄性C57BL/6小鼠随机分为对照组、PD模型组、40 Hz+PD干预组、PD+sh-NC组、PD+sh-STING组（n=8）。除对照组外，其余4组均采用腹腔注射MPTP（30 mg/kg）的方式建立亚急性PD小鼠模型；PD+sh-STING组进行脑立体定位注射AAV干扰STING基因；40 Hz+PD组进行连续4周的40 Hz光闪烁刺激干预。干预结束后通过行为学实验检测小鼠的运动能力与认知功能；采用免疫荧光染色观察小鼠海马S100钙结合蛋白A10（S100A10）、补体C3（C3）与星形胶质细胞蛋白GFAP的共定位水平；尼氏染色观察尼氏小体的数量；ELISA检测小鼠血清中的白介素含量；Western blotting和RT-qPCR分别检测黑质与海马组织中的α-突触核蛋白（α-syn）和酪氨酸羟化酶（TH）、突触可塑性、星形胶质细胞标志物、神经炎症、细胞因子信号传导抑制因子3（SOCS3）、JAK2-STAT3及STING通路相关蛋白与转录水平。结果与对照组相比，PD组小鼠运动能力与认知功能显著降低，TH蛋白表达降低（P<0.01），α-syn的表达增加（P<0.01）。40 Hz光闪烁刺激干预后PD小鼠认知功能得到改善，海马神经元数量显著增加（P<0.0001）、S100A10表达增加（P<0.05），p-STING/STING、C3、GFAP、IL-6、TNF-α表达减少（P<0.05）。与PD+sh-NC组小鼠相比，PD+sh-STING组小鼠认知功能明显恢复，海马区SOCS3、JAK2-STAT3表达水平无显著变化（P>0.05），而C3表达水平降低（P<0.05），S100A10表达水平则升高（P<0.05）。结论 40 Hz光闪烁刺激可能通过抑制STING信号通路的异常激活，促进星形胶质细胞向A2型极化，改善海马突触可塑性，进而缓解PD小鼠的认知功能障碍。

关键词: 40 Hz光闪烁刺激, 帕金森病, 认知功能障碍, 星形胶质细胞极化, STING信号

Abstract:

Objective To explore the effects of 40 Hz flicker light stimulation on cognitive dysfunction in mice with subacute Parkinson disease (PD) induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and the underlying mechanisms. Methods Forty 8-week-old male C57BL/6 mice were randomly allocated to a control group and 4 MPTP-induced PD model groups receiving blank treatment, 40 Hz flicker light stimulation, or stereotactic intraventricular injection of a control AAV (sh-NC) or an AVV carrying sh-STING. After the treatments, the mice were examined for changes in motor functions and cognitive behaviors. Immunofluorescence staining, Nissl staining, ELISA, Western blotting and RT-qPCR were used to detect hippocampal co-localization of S100A10 and C3 with GFAP, Nissl neuron count, serum interleukins, and nigral and hippocampal expressions of α-syn, TH, synaptic, astrocytic, inflammatory, SOCS3, JAK2-STAT3, and the STING pathway at the mRNA and protein levels. Results The PD mouse models exhibited impaired motor and cognitive functions with lowered TH and elevated α‑syn expressions. 40 Hz light flicker stimulation improved cognitive function of the PD mice, significantly increased the number of hippocampal neurons and S100A10 expression, and reduced the expressions of p-STING/STING, C3, GFAP, IL-6, and TNF‑α. Compared with the PD mice with sh-NC injection, the mice receiving sh-STING AVV injection showed significantly improved cognitive function with lowered hippocampal expression level of C3 and increased expression of S100A10 without significant changes in the expression levels of SOCS3 and JAK2-STAT3. Conclusion 40 Hz flicker light stimulation mitigates impairment of cognitive function in mice with MPTP-induced PD by inhibiting STING overactivation to promote A2 astrocyte polarization and enhance hippocampal synaptic plasticity.

Key words: 40 Hz light flicker stimulation, Parkinson disease, cognitive dysfunction, astrocyte polarization, STING signaling

任乾千, 张宝文, 刘醒然, 丁伟利, 陈丹丹, 寇现娟. 40 Hz光闪烁刺激通过抑制STING信号通路改善帕金森病小鼠的认知功能障碍[J]. 南方医科大学学报, 2026, 46(10): 2325-2337.

Qianqian REN, Baowen ZHANG, Xingran LIU, Weili DING, Dandan CHEN, Xianjuan KOU. 40 Hz flicker light stimulation suppresses STING signaling to improve cognitive dysfunction in mice with Parkinson disease[J]. Journal of Southern Medical University, 2026, 46(10): 2325-2337.

图/表 10

表1 引物序列

Tab.1 Primer sequence for RT-qPCR

Gene	Forward primer (5'-3')	Reverse primer (5'-3')
BDNF	TCATACTTCGGTTGCATGAAGG	AGACCTCTCGAACCTGCCC
SYN	AGCTCAACAAATCCCAGTCTCT	CGGATGGTCTCAGCTTTCAC
PSD93	ATCATTCCTTACCTCGGCTAACT	TCAGAGGAGAGATGTGAGACTG
PSD95	TGAGATCAGTCATAGCAGCTACT	CTTCCTCCCCTAGCAGGTCC
C1q	TGGAGACCTCTCAGGGATGG	CTTTCACGCCCTTCAGTCCT
Arg-1	CTCCAAGCCAAAGTCCTTAGAG	AGGAGCTGTCATTAGGGACATC
C3	TAGAGGCCCATGATGCTCAG	AAGTTTGCCACCACTGTCAC
S100A10	TGGAAACCATGATGCTTACGTT	GAAGCCCACTTTGCCATCTC
IL-6	TAGTCCTTCCTACCCCAATTTCC	TTGGTCCTTAGCCACTCCTTC
TNF-ɑ	GACGTGGAACTGGCAGAAGAG	TTGGTGGTTTGTGAGTGTGAG
GAPDH	GGTTGTCTCCTGCGACTTCA	TGGTCCAGGGTTTCTTACTCC

图1 PD小鼠造模以及运动功能检测结果

Fig.1 Modeling of Parkinson disease (PD) mice and motor function test results. A: Dark box residence time in the black-and-white box test. B: Duration time in the rotating rod test. C: Duration time in the hanging test. D: Pathway map and heat map of open field test. E: Total distance and movement speed in the open field test. F: Western blotting detection results. G: Expression levels of α-syn and TH proteins. Data are presented as Mean±SD and analyzed using one-way ANOVA. #P<0.05, ##P<0.01, ####P<0.0001 vs Control. *P<0.05, **P<0.01 vs PD.

图2 各组小鼠认知功能检测结果

Fig.2 Cognitive function test results in each group. A: Water maze test trajectory. B: Target quadrant activity distance within the water maze. C: Number of target platform crossing within the water maze. D: Y maze test trajectory and heat map. E: Y maze spontaneous response alternation rate. F: Novel object recognition index. Data are presented as Mean±SD. #P<0.05,##P<0.01,###P<0.001 vs Control. *P<0.05 vs PD.

图3 各组小鼠海马神经元及突触相关检测结果

Fig.3 Detection results of hippocampal neurons and synapses in each group. A: Nissl staining of the hippocampus in each group (Original magnification: × 50). B: Nissl neuronal count statistics in the hippocampus in each group. C: Western blotting detection results. D: Analysis of Western blotting detection results. E: Analysis of RT-qPCR detection results. Data are presented as Mean±SD and analyzed using one-way ANOVA. #P<0.05,##P<0.01, ###P<0.001,####P<0.0001 vs Control. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs PD.

图4 各组小鼠海马SOCS3以及JAK2-STAT3-STING通路检测结果

Fig.4 Detection results of SOCS3 and the JAK2-STAT3-STING pathway in mouse hippocampus in each group. A: Western blotting detection results. B: Analysis of the results of Western blotting. Data are presented as Mean±SD and analyzed using one-way ANOVA. #P<0.05,##P<0.01, ###P<0.001,####P<0.0001 vs Control. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs PD.

图5 各组小鼠海马星形胶质细胞极化表型检测结果

Fig.5 Detection results of hippocampal astrocyte polarization phenotypes in the mice in each group. A: Immunofluorescence staining of detecting hippocampal GFAP (green) and C3 (yellow) in each group (×100; arrows indicate co-staining regions of GFAP and C3). B: Quantitative analysis of immunofluorescence staining results. C: Immunofluorescence staining for detecting hippocampal GFAP (green) and S100A10 (red) in each group (×100; arrows indicate co-staining regions of GFAP and S100A10). D: Quantitative analysis of immunofluorescence staining results. E: Western blotting detection results. F: Quantitative analysis of Western blotting results. G: RT-qPCR detection analysis. Data are presented as Mean±SD and analyzed using one-way ANOVA. ##P<0.01, ###P<0.001,####P<0.0001 vs Control. *P<0.05, **P<0.01, ***P<0.001 vs PD.

图6 各组小鼠海马神经炎症及全身性炎症检测结果

Fig.6 Detection results of hippocampal neuroinflammation and systemic inflammation in each group. A: Analysis of RT-qPCR detection results. B: Serum ELISA detection results. C: Western blotting detection results. D: Quantitative analysis of Western blotting detection results. Data are presented as Mean±SD and analyzed using one-way ANOVA. ##P<0.01, ###P<0.001, ####P<0.0001 vs Control. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs PD.

图7 STING干扰小鼠认知功能检测结果

Fig.7 Effect of STING knockdown on cognitive function of PD mice. A: Western blotting detection results. B: Quantitative analysis of Western blotting results. C: Water maze test trajectory. D: Target quadrant activity path within the water maze. E: Number of target platform crossings within the water maze. F: Y maze test trajectory and heat map. G: Y maze spontaneous response alternation rate. Data are presented as Mean±SD and analyzed using one-way ANOVA. *P<0.05, **P<0.01, ***P<0.001 vs PD+sh-NC.

图8 STING干扰小鼠海马SOCS3-JAK2-STAT3信号通路检测结果

Fig.8 Detection results of the SOCS3-JAK2-STAT3 signaling pathway in the hippocampus of PD mice with STING knockdown. A: Western blotting detection results. B: Quantitative analysis of Western blotting results.

图9 STING干扰小鼠海马星形胶质细胞极化检测结果

Fig. 9 STING knockdown affects detection results of hippocampal astrocyte polarization in mice. A: Western blotting detection results. B: Quantitative analysis of Western blotting results. C: RT-qPCR for detection of C3. D: RT-qPCR for detection of S100A10. Data are presented as Mean±SD and analyzed using one-way ANOVA. *P<0.05, **P<0.01, ***P<0.001 vs sh-NC.

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40 Hz光闪烁刺激通过抑制STING信号通路改善帕金森病小鼠的认知功能障碍

40 Hz flicker light stimulation suppresses STING signaling to improve cognitive dysfunction in mice with Parkinson disease

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