电针调控VD大鼠HIF-1α/CXCR4/NF-κB和HO-1/SOD信号通路交互改善认知的机制研究[1]
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1.辽宁中医药大学针灸推拿学院;2.辽宁中医药大学中医脏象理论及应用教育部重点实验室;3.辽宁中医药大学附属第二医院;4.辽宁中医药大学实验动物医学院

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R245

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国家自然科学基金面上项目(82474634);辽宁省科技计划联合项目(2023-MSLH-178);


Mechanism Study of Electroacupuncture Regulation of HIF-1α/CXCR4/NF-κB and HO-1/SOD Signaling Pathway Interactions in VD Rats to Improve Cognitive Function

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1.College of Acupuncture and Tuina,Liaoning University of Traditional Chinese Medicine;2.Key Laboratory of Traditional Chinese Medicine Zang-Xiang Theory and Application,Ministry of Education,Liaoning University of Traditional Chinese Medicine;3.The Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, College of Experimental Animal Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang

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    摘要:

    目的:观察电针对血管性痴呆(VD)大鼠海马组织HIF-1α/CXCR4/NF-κB、HO-1/SOD通路的影响,探讨电针通过调控通路信号交互、抑制炎症反应、修复氧化应激损伤,改善认知障碍的潜在机制。方法:采用SPF级雄性SD大鼠作为实验对象,按随机数字表法分为假手术组、模型组、电针组和假电针组,采用双侧颈总动脉结扎法(2-VO)制备VD大鼠模型。电针组予“百会”“足三里”“三阴交”进行干预,15min /次,1次/d,共干预14d,假电针组将毫针浅刺皮下,不予通电,假手术组和模型组仅接受固定处理。干预结束后,采用Morris水迷宫及Y迷宫检测各组大鼠认知与记忆能力;HE染色法观察大脑海马区形态;实时荧光定量PCR法检测海马组织HIF-1α、CXCR4 mRNA的相对表达量;Western blot法检测海马组织HIF-1α、CXCR4蛋白的相对表达与p-NF-κB/NF-κB的比值;免疫组织化学法检测海马组织TNF-α、IL-6、IL-1β的平均吸光度;免疫荧光法检测大鼠海马组织HO-1的平均荧光强度;ELISA法检测海马组织MDA、SOD、ROS的含量;荧光探针法观察海马区ROS的积累情况。结果:与假手术组比较,模型组与假电针组大鼠逃避潜伏期延长、穿越平台次数减少、自发交替率降低(P<0.01),海马区神经元排列紊乱、松散,细胞间隙增大,海马组织HIF-1α、CXCR4 mRNA及蛋白的相对表达升高(P<0.01)、p-NF-κB/NF-κB比值升高(P<0.01),TNF-α、IL-6、IL-1β平均吸光度升高(P<0.01),HO-1荧光强度降低(P<0.01),MDA含量升高(P<0.05)、SOD含量降低(P<0.01),ROS含量与积累升高(P<0.05)。相较于模型组,电针组大鼠逃避潜伏期缩短、穿越平台次数增加、自发交替率升高(P<0.01),海马区损伤减轻,HIF-1α、CXCR4的mRNA及蛋白表达降低(P<0.01),p-NF-κB/NF-κB比值降低(P<0.01),TNF-α、IL-6、IL-1β平均吸光度降低(P<0.01),HO-1荧光强度升高(P<0.01),MDA含量降低(P<0.05)、SOD含量升高(P<0.01),ROS含量与积累降低(P<0.05)。结论:电针“百会”“足三里”“三阴交”可修复VD大鼠海马神经元氧化损伤,改善认知功能,其机制与平衡HIF-1α/CXCR4/NF-κB炎性通路和HO-1/SOD抗氧化通路的交互,减轻神经炎症,提高抗氧化应激水平有关。

    Abstract:

    Objective: To examine the impact of electroacupuncture on HIF-1α/CXCR4/NF-κB and HO-1/SOD pathways in hippocampal tissue of vascular dementia (VD) rats, and to explore the potential mechanisms by which electroacupuncture improves cognitive impairment through regulating pathway signal interactions, inhibiting inflammatory responses, and repairing oxidative stress damage. Methods: SPF-grade male SD rats were randomly assigned to sham surgery, model, electroacupuncture, and sham electroacupuncture groups. Vascular dementia (VD) was induced by bilateral common carotid artery ligation (2-VO). The electroacupuncture group received intervention at Baihui (GV10), Zusanli (ST36), and Sanyinjiao (SP6) points, administered at 15 min/session, once daily for 14 days. The sham electroacupuncture group received shallow subcutaneous needle insertion without electrical stimulation. The sham surgery and model groups underwent fixation procedures only. Following intervention, cognitive and memory abilities were assessed using the Morris water maze and Y-maze. Histomorphology of the hippocampal region was examined via hematoxylin and eosin (HE) staining. Relative mRNA expression of HIF-1α and CXCR4 in hippocampal tissue was quantified by qPCR. Western blot analysis determined relative protein expression of HIF-1α and CXCR4 alongside the p-NF-κB/NF-κB ratio in hippocampal tissue. Immunohistochemistry was used to measure the mean absorbance of TNF-α, IL-6, and IL-1β in hippocampal tissue.Immunofluorescence was performed to measure the mean fluorescence intensity of HO-1 in rat hippocampal tissue. ELISA was used to determine the levels of MDA, SOD, and ROS in hippocampal tissue. Fluorescent probes were utilized to observe ROS accumulation in the hippocampal region. Results: Relative to sham-operated controls, both the model group and the sham electroacupuncture group exhibited prolonged escape latency, reduced platform crossings, and decreased spontaneous alternation rate (P<0.01). Neuronal arrangement in the hippocampal region was disordered and loosened, with enlarged intercellular gaps. Relative expression of HIF-1α and CXCR4 mRNA and proteins in hippocampal tissue increased (P<0.01), the p-NF-κB/NF-κB ratio increased (P<0.01), increased mean absorbance of TNF-α, IL-6, and IL-1β (P<0.01), decreased HO-1 fluorescence intensity (P<0.01), elevated MDA content (P<0.05), reduced SOD content (P<0.01), and increased ROS content and accumulation (P<0.05). Compared with model rats, electroacupuncture-treated rats exhibited a shortened escape latency, increased platform crossings, and elevated spontaneous alternation rate (P<0.01). Hippocampal damage was reduced, with decreased mRNA and protein expression of HIF-1α and CXCR4 (P<0.01), a reduced p-NF-κB/NF-κB ratio (P<0.01), TNF-α, IL-6, and IL-1β mean optical densities decreased (P<0.01), HO-1 fluorescence intensity increased (P<0.01), MDA content decreased (P<0.05), SOD content increased (P<0.01), and ROS content and accumulation decreased (P<0.05). Conclusion: Electroacupuncture at Baihui (GV20), Zusanli (ST36), and Sanyinjiao (SP6) repaired oxidative damage to hippocampal neurons in VD rats and improved cognitive function. This effect was mediated by balancing the HIF-1α/CXCR4/NF-κB inflammatory pathway and the HO-1/SOD antioxidant pathway, thereby reducing neuroinflammation and enhancing antioxidant stress levels.

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韩磊,杨琳,王建波,等. 电针调控VD大鼠HIF-1α/CXCR4/NF-κB和HO-1/SOD信号通路交互改善认知的机制研究[1][J]. 科学技术与工程, , ():

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  • 收稿日期:2026-03-04
  • 最后修改日期:2026-05-09
  • 录用日期:2026-05-15
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