“嗅三针”调控小胶质细胞改善血管性痴呆大鼠髓鞘再生的机制研究  

Mechanism of"olfactory three needles"in regulating microglia and promoting remyelination in vasculardementia rats

作　　者：李泐 王强 刘隽阳 赵蔚佳 曾佳玮 张兵兵 茂瑞瑞 冯卫星 李杰 LI Le;WANG Qiang;LIU Junyang;ZHAO Weji;ZENG Jiawei;ZHANG Bingbing;MAO Ruirui;FENG Weixing;LI Jie(School of Acupuncture-Moxibustion and Tuina,Shaanxi University of CM,Xianyang 712046,China;Shaanxi Provincial Key Laboratory of Acupuncture and Medicine,Xianyang 712046;Second Clinical Medical College,Shaanxi University of CM;Affiliated Hospital of Shaanxi University of CM)

机构地区：[1]陕西中医药大学针灸推拿学院,咸阳712046 [2]陕西省针药结合重点实验室,咸阳712046 [3]陕西中医药大学第二临床医学院 [4]陕西中医药大学附属医院

出　　处：《中国针灸》2025年第4期473-481,共9页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金资助项目:82004480。

摘　　要：目的:观察“嗅三针”对血管性痴呆(VD)模型大鼠认知、学习记忆能力及海马小胶质细胞(MG)吞噬活性的影响,探讨针刺调节MG活化及髓鞘再生改善VD的机制。方法:从38只符合实验要求的SD大鼠中随机选取9只作为假手术组,其余大鼠采用永久性双侧颈动脉结扎法制备VD模型,造模成功的18只大鼠随机分为模型组、电针组,每组9只。电针组大鼠选取“嗅三针”(“印堂”及双侧“迎香”)进行电针干预,予疏密波,频率为2 Hz/15 Hz,电流1 mA,每次15 min,每日1次,7 d为一疗程,疗程间休息2 d,共2个疗程。采用新物体识别实验观察大鼠认知能力,Morris水迷宫实验观察大鼠学习记忆能力,LFB染色观察大鼠海马区髓鞘脱失情况,Western blot法检测大鼠海马区髓系细胞触发受体2(TREM2)、髓鞘蛋白脂质蛋白(PLP)蛋白表达,免疫荧光法检测大鼠海马PLP、性别决定区Y框蛋白10(SOX10)、离子钙结合适配器分子1(Iba1)^(+)TREM2^(+)、Iba1^(+)溶酶体关联膜蛋白1(LAMP1)^(+)阳性表达。结果:与假手术组比较,模型组大鼠第3、4天逃避潜伏期延长(P<0.05,P<0.01),总路程增加(P<0.01),认知指数(RI)、穿越平台次数降低(P<0.01);与模型组比较,电针组大鼠第3、4天逃避潜伏期缩短(P<0.05),总路程减少(P<0.01),穿越平台次数、RI升高(P<0.05,P<0.01)。与假手术组比较,模型组大鼠海马CA1区染色不均匀,髓鞘纤维排列稀疏,轮廓不清晰,出现较多空泡样改变;与模型组比较,电针组大鼠海马CA1区染色加深,髓鞘纤维增多、排列整齐,空泡样改变减少。与假手术组比较,模型组大鼠海马TREM2蛋白表达升高、PLP蛋白表达降低(P<0.01);与模型组比较,电针组大鼠海马TREM2、PLP蛋白表达升高(P<0.01,P<0.05)。与假手术组比较,模型组大鼠海马PLP、SOX10、Iba1+LAMP1+阳性表达降低(P<0.05,P<0.01),Iba1^(+)TREM2^(+)阳性表达升高(P<0.05);与模型组比较,电针组大鼠海马PLP、SOX10、Iba1^(+)TREM2Objective To observe the effects of"olfactory three needles"on cognition,learning and memory abilities,as well as hippocampal microglia(MG)phagocytic activity in vascular dementia(VD)rats,and explore the mechanisms of acupuncture in regulating (MG) activation and improving remyelination,so as to ameliorate VD.Methods Among 38 SD rats meeting experimental requirements,9 rats were randomly assigned to a sham-operation group,and the remaining rats underwent permanent bilateral common carotid artery ligation to establish VD model.Eighteen successfully modeled rats were randomly divided into a model group and an electroacupuncture(EA)group,with 9 rats in each one.In the EA group,EA was performed at"olfactory three needles"("Yintang"[GV24^(+)]and bilateral"Yingxiang"[LI20D]),at disperse-dense wave,the frequency of 2 Hz/15 Hz and the current intensity of 1 mA,for 15 min per intervention,once daily.One course was composed of 7 days,and 2 courses were required,with the interval of 2 days.The novel object recognition test was employed to assess the cognition of rats,and the Morris water maze was adopted to observe learning and memory abilities.Luxol fast blue(LFB)staining was performed to evaluate myelin sheath loss in the hippocampus,the Western blot was used to detect the protein expression of triggering receptor expressed on myeloid cells-2(TREM2)and proteolipid protein(PLP)in the hippocampus;and the immunofluorescence staining was used to detect the positive expression of PLP,sex determining region Y-box 10(SOX10),ionized calcium binding adaptor molecule 1(lbal)TREM2 and Ibal^(+)lysosome-associated membrane protein 1(LAMP1)in the hippocampus.Results Compared with the sham-operation group,the rats in the model group exhibited the prolonged escape latency on day 3 and 4(P<0.05,P<0.01),the increase of the total distance traveling(P<0.01)and the decrease of the recognition index(RI)and platform crossing frequency(P<0.01).Compared with the model group,the rats in the EA group showed the shortened escape latency on day 3 and

关 键 词：血管性痴呆 嗅三针 电针 小胶质细胞 少突胶质细胞 髓鞘 

分 类 号：R245[医药卫生—针灸推拿学]