| 引用本文: |
陈盼, 谢紫薇, 李娜, 黄超飞, 张泓, 阳庆军, 邹莹洁, 袁思, 谭洁.运动预处理结合电针对血管性痴呆大鼠海马小胶质细胞极化及TREM2/DAP12蛋白表达的影响[J].湖南中医药大学学报,2025,45(6):1044-1053[点击复制] |
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| 运动预处理结合电针对血管性痴呆大鼠海马小胶质细胞极化及TREM2/DAP12蛋白表达的影响 |
| 陈盼,谢紫薇,李娜,黄超飞,张泓,阳庆军,邹莹洁,袁思,谭洁 |
| (湖南中医药大学针灸推拿与康复学院, 湖南 长沙 410208;湘雅博爱康复医院康复治疗部, 湖南 长沙 410000) |
| 摘要: |
| 目的 观察运动预处理结合电针对血管性痴呆（VD）大鼠海马小胶质细胞极化、髓系细胞触发受体2（TREM2）/衔接蛋白DNAX激活蛋白12（DAP12）的影响，探讨运动预处理结合电针改善VD大鼠认知功能的可能机制。方法 将90只雄性SD大鼠随机均分为非运动预处理组和运动预处理组，非运动预处理组又均分为假手术组（Sham组）、模型组（Model组）和电针组（EA组），运动预处理组又均分为运动预处理假手术组（EP-Sham组）、运动预处理模型组（EP-Model组）和运动预处理电针组（EP-EA组），每组15只。运动预处理组所有大鼠在造模前行不负重游泳运动训练4周，每周5 d，每天30 min。Model组、EP-Model组、EA组和EP-EA组通过双侧颈总动脉永久性双重结扎法制备VD模型，Sham组和EP-Sham组只分离双侧颈总动脉不作结扎。造模后第7天，EP-EA组和EA组大鼠进行为期4周的电针治疗（电针百会、大椎和双侧肾俞），每周6 d，每天30 min。于运动预处理前、运动预处理后及电针干预后，采用Morris水迷宫测试大鼠学习记忆能力，罗克沙尔坚牢蓝染色观察大鼠海马CA1区髓鞘脱失情况，免疫荧光双标染色法检测大鼠海马CA1区诱导型一氧化氮合酶（iNOS）、精氨酸酶1（Arg1）与离子钙结合适配器分子1（Iba1）共表达水平，Western blot法检测大鼠海马组织中TREM2、DAP12的表达水平。结果 与Sham组比较，Model组大鼠平均逃避潜伏期延长、穿越平台次数减少（P＜0.05）；海马CA1区完整髓鞘数量明显降低，髓鞘组织海绵状空泡样改变严重，小胶质细胞胞质内可见大量髓鞘碎片；髓鞘脱失评分显示海马CA1区髓鞘脱失增多（P＜0.05）；海马CA1区中iNOS与小胶质细胞标志物Iba1共表达水平增加（P＜0.05），Arg1与小胶质细胞标志物Iba1共表达水平减少（P＜0.05）；海马中TREM2、DAP12蛋白表达下降（P<0.05）。与Model组比较，EA组、EP-Model组和EP-EA组大鼠平均逃避潜伏期缩短、穿越平台次数增多（P＜0.05）；海马CA1区髓鞘较为完整，排列较为整齐和均匀；髓鞘脱失评分显示海马CA1区髓鞘脱失减少（P＜0.05）；海马CA1区中iNOS与Iba1共表达水平减少（P＜0.05），Arg1与Iba1共表达水平增加（P＜0.05）；海马中TREM2、DAP12蛋白表达升高（P<0.05）。与EP-Model组比较，EP-EA组大鼠平均逃避潜伏期缩短、穿越平台次数增多（P<0.05）；髓鞘脱失评分显示海马CA1区髓鞘脱失减少（P<0.05）；大鼠海马CA1区中iNOS与Iba1共表达水平减少（P＜0.05），Arg1与Iba1共表达水平增加（P<0.05）；海马中TREM2、DAP12蛋白表达升高（P<0.05）。结论 运动预处理结合电针可改善VD大鼠的学习和记忆功能，其机制可能是通过调控TREM2、DAP12蛋白表达，促进海马小胶质细胞极化为M2型，增强M2型小胶质细胞吞噬功能，促进髓鞘碎片的清除。 |
| 关键词: 血管性痴呆 小胶质细胞极化 运动预处理 电针 髓系细胞触发受体2 衔接蛋白DNA X激活蛋白12 |
| DOI：10.3969/j.issn.1674-070X.2025.06.008 |
| 投稿时间：2024-11-29 |
| 基金项目:湖南省教育厅科学研究重点项目（22A0253）；湖南中医药大学研究生创新课题（2023CX164）。 |
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| Effects of exercise pretreatment combined with electroacupuncture on microglial polarization and TREM2/DAP12 protein expression in the hippocampus of rats with vascular dementia |
| CHEN Pan, XIE Ziwei, LI Na, HUANG Chaofei, ZHANG Hong, YANG Qingjun, ZOU Yingjie, YUAN Si, TAN Jie |
| (College of Acupuncture-Moxibustion-Tuina and Rehabilitation, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Department of Rehabilitation, Xiangya Boai Rehabilitation Hospital, Changsha, Hunan 410000, China) |
| Abstract: |
| Objective To investigate the effects of exercise pretreatment combined with electroacupuncture on hippocampal microglial polarization and triggering receptor expressed on myeloid cells 2 (TREM2)/DNAX-activating protein 12 (DAP12) in vascular dementia (VD) rats, and to explore the possible mechanisms by which exercise pretreatment combined with electroacupuncture improves cognitive function in VD rats.Methods Ninety male SD rats were randomly divided into non-exercise pretreatment group and exercise pretreatment group. The non-exercise pretreatment group was further divided into sham-operated group (Sham group), model group, and electroacupuncture group (EA group). The exercise pretreatment group was further divided into exercise pretreatment-sham operation group (EP-Sham group), exercise pretreatment-model group (EP-Model group), and exercise pretreatment-electroacupuncture group (EP-EA group), with 15 rats in each group. All rats in the exercise pretreatment group were subjected to unloaded swimming exercise training for 4 weeks prior to model establishment, with training being conducted for 5 d per week and lasting 30 minutes each day. VD models were established in the model group, EP-Model group, EA group, and EP-EA group through permanent bilateral common carotid artery double ligation, while in the Sham group and EP-Sham group, only the bilateral common carotid arteries were isolated without ligation. On the 7th day after model establishment, rats in the EP-EA group and EA group were subjected to a 4-week course of electroacupuncture treatment [electroacupuncture at "Baihui (GV20)", "Dazhui (GV14)", and bilateral "Shenshu (BL23)" acupoints], with the treatment being administered 6 d a week, 30 minutes each day. Morris water maze test was used to test the learning and memory abilities of rats before exercise pretreatment, after exercise pretreatment and after electroacupuncture intervention. Luxol fast blue staining was used to observe the demyelination in the hippocampal CA1 region of the rats. Immunofluorescence double staining was used to measure the co-expression levels of inducible nitric oxide synthase (iNOS), arginase 1 (Arg1), and ionized calcium-binding adapter molecule 1 (Iba1) in the hippocampal CA1 region of the rats. Western blot was used to measure the expression levels of TREM2 and DAP12 in the hippocampal tissue of the rats.Results Compared with the Sham group, the model group rats showed prolonged mean escape latency and decreased number of platform crossings (P<0.05). The number of intact myelin sheaths in the hippocampal CA1 region significantly reduced, with severe spongiform vacuolar changes in the myelin sheath tissue and a large amount of visible myelin debris in the cytoplasm of microglia. The myelin loss score indicated an increase in myelin loss in the hippocampal CA1 region (P<0.05). The co-expression levels of iNOS and the microglial marker Iba1 in the hippocampal CA1 region increased (P<0.05), while those of Arg1 and Iba1 decreased (P<0.05). The protein expressions of TREM2 and DAP12 in the hippocampus decreased (P<0.05). Compared with the model group, rats in the EA, EP-Model, and EP-EA groups showed shortened mean escape latency and increased number of platform crossings (P<0.05). The myelin sheaths in the hippocampal CA1 region were relatively intact, with comparatively regular and uniform arrangement. The myelin loss score indicated a decrease in myelin loss in the hippocampal CA1 region (P<0.05). The co-expression levels of iNOS and Iba1 in the hippocampal CA1 region decreased (P<0.05), while those of Arg1 and Iba1 increased (P<0.05). The protein expressions of TREM2 and DAP12 in the hippocampus were elevated (P<0.05). Compared with the EP-Model group, the EP-EA group rats showed shortened mean escape latency and increased number of platform crossings (P<0.05). The myelin loss score indicated a decrease in myelin loss in the hippocampal CA1 region (P<0.05). The co-expression levels of iNOS and Iba1 in the hippocampal CA1 region decreased (P<0.05), while those of Arg1 and Iba1 increased (P<0.05). The protein expressions of TREM2 and DAP12 in the hippocampus were elevated (P<0.05).Conclusion Exercise pretreatment combined with electroacupuncture can improve learning and memory functions in VD rats, and the underlying mechanism may involve regulating the TREM2 and DAP12 protein expressions, promoting the polarization of hippocampal microglia toward the M2 phenotype, enhancing the phagocytic capacity of M2 microglia, and facilitating the clearance of myelin debris. |
| Key words: vascular dementia microglial polarization exercise pretreatment electroacupuncture triggering receptor expressed on myeloid cells 2 DNAX-activating protein 12 |
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