电针改善血管性痴呆大鼠认知障碍并抑制TLR4/MyD88/NF-κB通路介导的星形胶质细胞活化

doi:10.12122/j.issn.1673-4254.2026.08.09

摘要/Abstract

摘要：

目的探讨电针对血管性痴呆（VD）大鼠认知功能及神经炎症反应的影响，并观察其对Toll样受体4/髓样分化初级反应蛋白88/核因子κB（TLR4/MyD88/NF-κB）通路介导星形胶质细胞异常活化的调控作用。方法 60只SPF级雄性SD大鼠随机分为假手术组（n=10）和造模组（n=50）。采用双侧颈总动脉结扎术（2-VO）制备VD模型，筛选造模成功大鼠30只，随机分为模型组、电针组及西药组（每组10只）。电针组选取“百会”、“神庭”穴，采用疏密波（2 Hz/15 Hz，1 mA，30 min/d）干预；西药组灌胃盐酸多奈哌齐（0.45 mg/kg），连续治疗28 d。通过Morris水迷宫评估认知功能；苏木精-伊红和尼氏染色观察神经元病理损伤；免疫组织化学法及透射电子显微镜检测胶质纤维酸性蛋白（GFAP）标记的星形胶质细胞活化状态及超微结构；免疫荧光检测GFAP与磷酸化NF-κB（p-NF-κB）共定位；ELISA测定海马炎症因子白细胞介素1β（IL-1β）、白细胞介素6（IL-6）和肿瘤坏死因子α（TNF-α）水平；Western blotting检测补体成分3（C3）、S100钙结合蛋白A10（S100A10）、TLR4、MyD88蛋白表达及p-NF-κB/NF-κB比值。结果与假手术组相比，模型组大鼠逃避潜伏期延长、平台穿越次数减少、目标象限停留时间缩短（P<0.01）；海马神经元排列紊乱、核固缩；星形胶质细胞呈异常激活状态，超微结构受损，GFAP与p-NF-κB共定位表达增强；炎症因子水平、C3及TLR4/MyD88/NF-κB通路蛋白表达均显著升高（P<0.01），S100A10的表达量显著降低（P<0.01）。与模型组相比，电针与西药干预均能显著缩短逃避潜伏期，增加平台穿越次数（P<0.01）；减轻神经元病理损伤，抑制星形胶质细胞过度活化及超微结构破坏；降低促炎因子含量，下调C3、TLR4、MyD88蛋白表达及p-NF-κB/NF-κB比值（P<0.05，P<0.01），上调S100A10的表达（P<0.05，P<0.01）。结论电针“神庭”、“百会”可改善VD大鼠认知障碍，减轻神经炎症反应，其作用机制可能与下调TLR4/MyD88/NF-κB通路相关蛋白表达、调节星形胶质细胞A1/A2样表型失衡有关。

关键词: 血管性痴呆, 电针, 星形胶质细胞, 神经炎症, TLR4/MyD88/NF-κB通路

Abstract:

Objective To investigate the effects of electroacupuncture (EA) on cognitive function and neuroinflammation in a rat model of vascular dementia (VD) and the underlying mechanism. Methods Sixty male SD rats were randomly assigned to sham-operated group (n=10) and VD model group (n=50) receiving bilateral common carotid artery occlusion. Thirty rats with successful VD modeling were randomized into model group, EA group, and donepezil treatment group (n=10). EA treatment was administered at the acupoints Baihui (GV20) and Shenting (GV24) with a disperse-dense wave (2/15 Hz, 1 mA, 30 min/day), and donepezil was given by gavage at 0.45 mg/kg. Both interventions lasted 28 days. Cognitive function of the rats was assessed using Morris water maze test, and neuronal pathologies were observed using HE and Nissl staining. GFAP-labeled astrocyte activation was assessed by immunohistochemistry, and astrocytic ultrastructure was examined with transmission electron microscopy. GFAP/p-NF-κB colocalization was detected by immunofluorescence staining. Hippocampal IL-1β, IL-6, and TNF-α levels were measured by ELISA, and the protein expression levels of C3, S100A10, TLR4, and MyD88 and the p-NF-κB/NF‑κB ratio were detected by Western blotting. Results Compared with the sham-operated rats, VD rats showed significant cognitive impairment, obvious neuronal disorganization and pyknosis in the hippocampus, excessive astrocyte activation, increased GFAP/p-NF‑κB colocalization, inflammatory cytokine levels and expressions of C3 and TLR4/MyD88/NF-κB pathway proteins, and decreased expression of S100A10. Treatment with EA and donepezil significantly improved the performance of the rats in Morris water maze test, alleviated neuronal injury, inhibited astrocyte overactivation and ultrastructural damage, reduced inflammatory cytokine levels, expressions of C3, TLR4, and MyD88 proteins and the p-NF-κB/NF-κB ratio, and increased the expression of S100A10 in the hippocampus. Conclusion EA at GV20 and GV24 improves cognitive impairment and attenuate neuroinflammation in VD rats possibly by inhibiting TLR4/MyD88/NF-κB signaling and regulating astrocytic A1/A2-like phenotypic imbalance.

Key words: vascular dementia, electroacupuncture, astrocytes, neuroinflammation, TLR4/MyD88/NF-κB pathway

高建红, 史册, 李蔚然, 尚祥, 王飞, 杨琪琪, 李飞. 电针改善血管性痴呆大鼠认知障碍并抑制TLR4/MyD88/NF-κB通路介导的星形胶质细胞活化[J]. 南方医科大学学报, 2026, 46(8): 1814-1822.

Jianhong GAO, Ce SHI, Weiran LI, Xiang SHANG, Fei WANG, Qiqi YANG, Fei LI. Electroacupuncture ameliorates cognitive impairment and suppresses TLR4/MyD88/NF-κB pathway-mediated astrocyte activation in rats with vascular dementia[J]. Journal of Southern Medical University, 2026, 46(8): 1814-1822.

图/表 9

图1 Morris水迷宫结果比较

Fig.1 Comparison of performance of the rats in Morris water maze test (Mean±SD, n=10). **P<0.01 vs Sham group; ##P<0.01 vs Model group.

图2 病理形态学比较

Fig.2 Comparison of pathomorphology of the hippocampal neurons of the rats (HE staining).

图3 尼氏小体比较

Fig.3 Comparison of Nissl bodies in the hippocampus of the rats (Nissl staining, n=4).

图4 GFAP阳性表达比较

Fig.4 Comparison of GFAP expression in the hippocampus of the rats (Immunohistochemistry, n=4).

图5 各组大鼠星形胶质细胞超微结构比较

Fig.5 Ultrastructure of the astrocytes in the hippocampus of the rats (Transmission electron microscopy, A: original magnification: ×10 000; B: ×20 000).

图6 GFAP与p-NF-κB共定位表达比较

Fig.6 Colocalization of glial fibrillary acidic protein (GFAP) and phosphorylated nuclear factor-κB (p-NF-κB) in the hippocampus of the rats (Immunofluorescence staining).

图7 IL-1β、IL-6和TNF-α含量比较

Fig.7 Levels of IL-1β, IL-6 and TNF-α in the hippocampus of the rats (Mean±SD, n=3). **P<0.01 vs Sham group; ##P<0.01 vs Model group.

图8 C3、S100A10蛋白表达

Fig.8 Expression of C3 and S100A10 proteins in the hippocampus of the rats (Mean±SD, n=3). **P<0.01 vs Sham group; #P<0.05, ##P<0.01 vs Model group.

图9 TLR4、MyD88蛋白表达和p-NF-κB/NF-κB值比较

Fig.9 TLR4 and MyD88 protein expressions and the p-NF‑κB/NF‑κB ratio in the hippocampus of the rats (Mean±SD, n=3). **P<0.01 vs Sham group; #P<0.05, ##P<0.01 vs Model group.

参考文献 30

[1]	Rajeev V, Fann DY, Dinh QN, et al. Pathophysiology of blood brain barrier dysfunction during chronic cerebral hypoperfusion in vascular cognitive impairment[J]. Theranostics, 2022, 12(4): 1639-58. doi：10.7150/thno.68304
[2]	Wang WZ, Qiao O, Ji HX, et al. Autophagy in vascular dementia and natural products with autophagy regulating activity[J]. Pharmacol Res, 2021, 170: 105756. doi：10.1016/j.phrs.2021.105756
[3]	Mok VCT, Cai Y, Markus HS. Vascular cognitive impairment and dementia: Mechanisms, treatment, and future directions[J]. Int J Stroke, 2024, 19(8): 838-56. doi：10.1177/17474930241279888
[4]	Kim HJ, Kim H, Lee JH, et al. Toll-like receptor 4 (TLR4): new insight immune and aging[J]. Immun Ageing, 2023, 20(1): 67. doi：10.1186/s12979-023-00383-3
[5]	Qiao CM, Tan LL, Ma XY, et al. Mechanism of S100A9-mediated astrocyte activation via TLR4/NF-κB in Parkinson's disease[J]. Int Immunopharmacol, 2025, 146: 113938. doi：10.1016/j.intimp.2024.113938
[6]	Zou HH, Chen XR, Lu JC, et al. Neurotropin alleviates cognitive impairment by inhibiting TLR4/MyD88/NF-κB inflammation signaling pathway in mice with vascular dementia[J]. Neurochem Int, 2023, 171: 105625. doi：10.1016/j.neuint.2023.105625
[7]	丁旭, 邓祥敏, 殷紫, 等. 鸢尾黄素通过调节TLR4/MyD88/NF-κB信号通路改善血管性痴呆大鼠认知缺陷[J]. 中国免疫学杂志, 2024, 40(3): 540-5.
[8]	Wang XL, Liu XX, Zhang WY, et al. Acupuncture reduces neuroinflammation and apoptosis, regulates peripheral immunity, and modulates T-cell subset distribution in vascular dementia rats[J]. Chin Med, 2025, 20(1): 203. doi：10.1186/s13020-025-01243-6
[9]	文林月, 杨丽芳, 欧小燕, 等. 针刺通过良性调控反应性星形胶质细胞极化改善后循环缺血性眩晕[J]. 上海针灸杂志, 2024, 43(10): 1139-46. doi：10.13460/j.issn.1005-0957.2024.13.4006
[10]	史晓萍, 宗阿南, 陶钧, 等. 《关于善待实验动物的指导性意见》的研究[J]. 中国医科大学学报, 2007, 36(4):493.
[11]	田朝禧, 高建红, 黄静, 等. 延龄草总皂苷调控线粒体自噬对血管性痴呆大鼠的神经保护作用[J]. 解放军医学杂志, 2026, 51(3): 410-8.
[12]	Su Y, Zhong YY, Yuan NN, et al. Danggui-Shaoyao-San modulates sphingolipid metabolism to promote oligodendrocyte differentiation and maturation in vascular dementia rats[J]. Chin Med, 2026, 21(1): 11. doi：10.1186/s13020-025-01284-x
[13]	孙晓丽, 张锴镔, 郭淑贞, 等. 龙生蛭胶囊对血管痴呆大鼠认知功能的影响及作用机制[J]. 中国实验方剂学杂志, 2022, 28(9): 72-9.
[14]	中国针灸学会. 实验动物常用穴位名称与定位第2部分: 大鼠[J]. 针刺研究, 2021, 46(4): 351-2.
[15]	高建红, 陈雅雯, 田朝禧, 等. 延龄草总皂苷通过调控Shh信号通路减轻血管性痴呆大鼠神经元损伤[J]. 中国病理生理杂志, 2025, 41(2): 354-61.
[16]	Ma CM, Zhou Y, Yi W, et al. Electroacupuncture of the Baihui and Shenting acupoints for vascular dementia in rats through the miR-81/IL-16/PSD-95 pathway[J]. Ann Transl Med, 2022, 10(10): 540. doi：10.21037/atm-22-2068
[17]	Yang YH, Rao T, Jiang YJ, et al. Electroacupuncture ameliorates cognitive impairment and white matter injury in vascular dementia rats via activating HIF-1α/VEGF/VEGFR2 pathway[J]. Neuro-science, 2025, 573: 364-80. doi：10.1016/j.neuroscience.2025.03.063
[18]	Jurcau A, Simion A. Neuroinflammation in cerebral ischemia and ischemia/reperfusion injuries: from pathophysiology to therapeutic strategies[J]. Int J Mol Sci, 2022, 23(1): 14. doi：10.3390/ijms23010014
[19]	Xiao HH, Zhang FR, Li S, et al. Xinshubao tablet rescues cognitive dysfunction in a mouse model of vascular dementia: Involvement of neurogenesis and neuroinflammation[J]. Biomed Pharmacother, 2024, 172: 116219. doi：10.1016/j.biopha.2024.116219
[20]	Fan YY, Huo J. A1/A2 astrocytes in central nervous system injuries and diseases: Angels or Devils?[J]. Neurochem Int, 2021, 148: 105080. doi：10.1016/j.neuint.2021.105080
[21]	张程程, 张欣雨, 刘璐. 神经炎症疾病中星形胶质细胞和小胶质细胞互作机制的研究进展[J]. 中国免疫学杂志, 2025, 41(11): 2756-63.
[22]	Ahremenko E, Andreev A, Apushkin D, et al. Glial cells in the early stages of neurodegeneration: pathogenesis and therapeutic targets[J]. Int J Mol Sci, 2025, 26(24): 11995. doi：10.3390/ijms262411995
[23]	Saggu R, Schumacher T, Gerich F, et al. Astroglial NF-kB contributes to white matter damage and cognitive impairment in a mouse model of vascular dementia[J]. Acta Neuropathol Commun, 2016, 4(1): 76. doi：10.1186/s40478-016-0350-3
[24]	Zhao YX, Huang YY, Cao Y, et al. Astrocyte-mediated neuroinflammation in neurological conditions[J]. Biomolecules, 2024, 14(10): 1204. doi：10.3390/biom14101204
[25]	李方存, 张鼎, 刘迟, 等. 温肺降浊方基于miR-93和miR-330-5p介导的TLR4/MyD88/NF-κB信号通路改善血管性痴呆大鼠海马区病变[J]. 中国老年学杂志, 2025, 45(8): 1882-8.
[26]	Ransohoff RM. How neuroinflammation contributes to neuro-degeneration[J]. Science, 2016, 353(6301): 777-83. doi：10.1126/science.aag2590
[27]	Gao YR, Hu M, Niu XL, et al. Dl-3-n-butylphthalide improves neuroinflammation in mice with repeated cerebral ischemia-reperfusion injury through the Nrf2-mediated antioxidant response and TLR4/MyD88/NF‑κB signaling pathway[J]. Oxid Med Cell Longev, 2022, 2022: 8652741. doi：10.1155/2022/8652741
[28]	Jiang HS, Yang XY, Wang YZ, et al. Vitamin D protects against traumatic brain injury via modulating TLR4/MyD88/NF‑κB pathway-mediated microglial polarization and neuroinflammation[J]. Biomed Res Int, 2022, 2022: 3363036. doi：10.1155/2022/3363036
[29]	Zou XS, Shi L, Yin HL, et al. Compound Gaoziban tablet alleviates depression toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-kappa B pathway[J]. J Tradit Chin Med, 2022, 42(6): 956-64.
[30]	Bu Y, Li WS, Lin J, et al. Electroacupuncture attenuates immune-inflammatory response in hippocampus of rats with vascular dementia by inhibiting TLR4/MyD88 signaling pathway[J]. Chin J Integr Med, 2022, 28(2): 153-61. doi：10.1007/s11655-021-3350-5