电针智三针调节酪氨酸激酶受体的配体通路改善血管性痴呆的作用机制

doi:10.16098/j.issn.0529-1356.2023.06.010

摘要/Abstract

摘要：

目的观察电针智三针调节促红细胞生成素产生肝细胞受体B2/促红细胞生成素产生肝细胞配体B2/大丝裂原活化蛋白激酸1（EphB2/EphrinB2/BMK1）信号通路改善血管性痴呆大鼠神经损伤的潜在机制。方法 SD雄性成年大鼠80只，随机分为假手术组、模型组、非穴位电针组、尼莫地平组和电针智三针组，改良Pulsinelli四血管阻断法制作血管性痴呆大鼠模型，分组治疗后进行各组大鼠水迷宫实验，HE染色、Nissl染色和透射电子显微镜（TEM）观察海马CA1区病理变化，免疫组织化学法检测大鼠海马CA1区EphB2和BMK1表达，免疫印迹法检测大鼠海马CA1区EphB2和BMK1蛋白的表达情况。结果相比模型组，给予电针和尼莫地平治疗的血管性痴呆大鼠逃避潜伏期下降（P<0.05），而穿越平台的次数明显提高（P<0.05）；HE染色、Nissl染色和TEM结果显示，与模型组比较，电针智三针组、尼莫地平组大鼠的海马CA1区神经元排列较整齐，细胞及细胞器形态结构较完整；免疫组织化学法和免疫印迹法检测显示，与假手术组比较，模型组大鼠海马CA1区EphB2和BMK1的表达差异明显下降，差异有统计学意义（P<0.05）；与模型组比较，电针智三针组大鼠海马CA1区EphB2和BMK1的表达明显增加（P<0.05），非穴位电针组大鼠海马CA1区EphB2和BMK1的表达差异无统计学意义（P＞0.05），与尼莫地平组相比，电针智三针组大鼠海马CA1区EphB2和BMK1的表达明显增加（P<0.05）。结论电针智三针可能通过增加海马CA1区EphB2和BMK1的表达，改善血管性痴呆大鼠海马神经元的损伤，从而达到改善血管性痴呆大鼠的学习记忆。

关键词: 电针智三针, 血管性痴呆, 海马, 信号通路, 免疫印迹法, 大鼠

Abstract:

Objective To observe the potential mechanism of electroacupuncture regulating the erythropoietin-producing hepatocellular receptor B2/erythropoietin-producing hepatocellular receptor-interacting B2/big mitogen-activated protein kinase 1(EphB2/EphrinB2/BMK1) signaling pathway to improve neural damage in vascular dementia rats. Methods Eighty SD male adult rats were randomly divided into a sham surgery group, a model group, a non acupoint electroacupuncture group, a nimodipine group, and an electroacupuncture three needle group. The vascular dementia rat model was made by the modified Pulsinelli four vessel occlusion method. After grouping, the rats in each group were subjected to water maze test, HE staining, Nissl staining, and transmission electron microscopy(TEM) to observe the pathological changes in the hippocampal CA1 area, and the expression of EphB2 and BMK1 in the hippocampal CA1 area was detected by immunohistochemistry; Detection of EphB2 and BMK1 protein expression in rat hippocampal CA1 region was detected by Western blotting. Results Compared with the model group, the escape latency of vascular dementia rats treated with electroacupuncture and nimodipine decreased (P<0.05), while the number of crossing platforms significantly increased (P<0.05); The result of HE staining, Nissl staining and TEM showed that compared with the model group, the neurons in hippocampal CA1 area of rats in the EA Zhisanzhen group and nimodipine group were arranged orderly, and the morphology and structure of cells and organelle were complete; Immunohistochemical method and Western blotting showed that the expression of EphB2 and BMK1 in the CA1 region of hippocampus in the model group was significantly lower than that in the sham surgery group (P<0.05); Compared with the model group, the expression of EphB2 and BMK1 in the hippocampal CA1 region of rats in the electroacupuncture Zhisanzhen group significantly increased (P<0.05), while the expression of EphB2 and BMK1 in the hippocampal CA1 region of rats in the non acupoint electroacupuncture group was not statistically significant (P>0.05). Compared with the nimodipine group, the expression of EphB2 and BMK1 in the hippocampal CA1 region of rats in the electroacupuncture Zhisanzhen group significantly increased (P<0.05). Conclusion Electroacupuncture may improve the damage of hippocampal neurons in vascular dementia rats by increasing the expression of EphB2 and BMK1 in the CA1 region of the hippocampus, thereby improving the learning and memory of vascular dementia rats.

Key words: Electroacupuncture wisdom three needle, Vascular dementia, Hippocampus, Signal pathway, Western blotting, Rat

中图分类号:

R322

李霞朱世杰唐中生罗亚非范瑞娟谢高宇寇云芳陆莹 . 电针智三针调节酪氨酸激酶受体的配体通路改善血管性痴呆的作用机制[J]. 解剖学报, 2023, 54(6): 689-694.

LI Xia ZHU Shi-jie TANG Zhong-sheng LUO Ya-fei FAN Rui-juan XIE Gao-yu KOU Yun-fang LU Ying. Mechanism of electroacupuncture regulating the ligand pathway of tyrosine kinase receptor in improving vascular dementia[J]. Acta Anatomica Sinica, 2023, 54(6): 689-694.

参考文献

［1］Santos MAO, Bezerra LS, Correia CdC, et al.Neuropsycheatric symptoms invascular dementia:Epidemiologic and clicnical aspects［J］．Dement Neuropsychol,2018,12(1):40-44.

［2］Zhao WM. Study on the correlation between hemorheology indexes and vascular dementia ［J］. Chinese Journal of Applied Neurology, 2016,19 (2): 79-81. (in Chinese)

赵伟苗.血液流变学指标与血管性痴呆的相关性研究［J］.中国实用神经疾病杂志，2016,19(2):79-81.

［3］Murai KK, Pasquale EB. ‘Eph’ective signaling:forward, reverse and crosstalk［J］. J Cell Sci,2003, 116(Pt 14): 2823-2832.

［4］Hu R, Wei P, Jin L,et al.Overexpression of EphB2 in hippocampus rescues impaired NMDA receptors trafficking and cognitive dysfunction in Alzheimer model［J］. Cell Death Dis, 2017，8(3):e2717.

［5］Biederer T,Stagi M.Singnaling by synaptogenic molecules［J］.Curr Opin Neurobiol,2008,18(3):261-269.

［6］Suzaki Y, Yoshizumi M, Kagami S,et al.Hydrogen peroxide stimulates c-Src-mediated big mitogen-activated protein kinase 1 (BMK1) and the MEF2C signaling pathway in PC12 cells: potential role in cell survival following oxidative insults［J］. J Biol Chem，2002,277（11）: 9614-9621.

［7］Li HL, Xiang J, Ouyang LZh, et al. The effect of electroacupuncture on synaptic remodeling related factors on the EphrinB2/EphB2 signaling pathway in rats with middle cerebral artery infarction ［J］. Chinese Journal of Rehabilitation Medicine, 2016, 31 (10): 1067-1077. (in Chinese)

李洪亮，向娟，欧阳里知，等.电针对大脑中动脉梗塞大鼠EphrinB2/EphB2信号通路上突触重塑相关因子的影响［J］.中国康复医学杂志，2016，31(10)：1067-1077.

［8］Lin J, Wei YW, Tang ZhSh，et al.Effects of electroacuncture Zhi san on learning and memory ability and expressionof EphA4/ephrinA3 protein in vascular dementiaRates［J］.Asia Pacific Traditional Medicine, 2020,16 (9): 38-42. (in Chinese)

林吉，魏宇唯，唐中生，等.电针智三针对血管性痴呆大鼠学习记忆及EphA4/ephrinA3蛋白表达的影响［J］.亚太传统医药,2020,16(9):38-42.

［9］Shao YL, Zheng LX, Yang ShL.New progress in treatment of vascular dementia with traditional Chinese medicine［J］.China Medical Journal, 2019,16 (21): 39-42. (in Chinese)

邵亚兰,郑露茜,杨树龙.血管性痴呆的中医药治疗新进展［J］.中国医药导报,2019,16(21):39-42.

［10］Liu WH.Thinking on the development direction of acupuncture-moxibustion science from the study of genres of acupuncture and moxibustion［J］.Chinese J Acupuncture and Moxibustion, 2021,41 (9): 951-978. (in Chinese)

刘炜宏.从针灸流派研究思考针灸学发展方向［J］.中国针灸,2021,41(9):951-978.

［11］Yang T, Wang JJ, Cheng HL.Progress in experimental research on acquisition and moxibustion treatment of vascular dementia liaoning［J］.Journal of Liaoning University of Traditional Chinese Medicine, 2020,22 (12): 110-113.

杨涛,王婧吉,程红亮.针灸治疗血管性痴呆实验研究进展［J］.辽宁中医药大学报,2020,22(12):110-113.

［12］Jin ChF, Liang QCh, Wu Y, et al. The relationship between cognitive impairment and vascular dementia in the acute phase of first-episode cerebral infarction ［J］. Stroke and Nervous Diseases, 2013, 20 (5): 294-296. (in Chinese)

靳春凤，梁庆成，吴云，等．首发脑梗死急性期伴认知障碍与血管性痴呆的关系［J］．卒中与神经疾病，2013，20(5)：294-296.

［13］Zhang JG, Tang C,Liao WY,et al.The antiapoptotic and antioxidative stress effects of Zhisanzhen in the Alzheimer’s disease model rat［J］.Neuroreport, 2019,30(9):628-636.

［14］Saha N, Robev D, Mason EO,et al.Therapeutic potential of targeting the Eph/ephrin signaling complex［J］.Int J Biochem Cell Biol, 2018,105:123-133.

［15］Li HL, Xiang J, Ouyang LZh, et al. The effect of electroacupuncture on synaptic remodeling related factors on the EphrinB2/EphB2 signaling pathway in rats with middle cerebral artery infarction ［J］. Chinese Journal of Rehabilitation Medicine, 2016,31 (10): 1067-1077. (in Chinese)

李洪亮,向娟,欧阳里知,等.电针对大脑中动脉梗塞大鼠EphrinB2/EphB2信号通路上突触重塑相关因子的影响［J］.中国康复医学杂志,2016,31(10):1067-1077.

［16］Chen YZh, Liu Ch, Shi XD. The role of Eph family proteins in neuropsychiatric disorders such as Alzheimer’s disease ［J］. Chinese Journal of Biochemistry and Molecular Biology, 2018,34 (11): 1160-1165. (in Chinese)

陈永庄，刘超，石小东.Eph家族蛋白在阿尔茨海默症等神经精神疾病中的作用［J］.中国生物化学与分子生物学报, 2018, 34(11): 1160-1165.

［17］Darling TK, Lamb TJ. Emerging roles for eph receptors and ephrin ligands in immunity［J］.Front Immunol, 2019, 10: 1473.

［18］Kania A, Klein R.Mechanisms of ephrinEph signalling in development, physiology and disease［J］. Nat Rev Mol Cell Biol,2016, 17（4）:240-256.

［19］Atapattu L, Lackmann M, Janes PW.The role of proteases in regulating Eph/ephrin signaling ［J］.Cell Adh Migr,2014,8(4):294-307.

［20］Arvanitis DN, Davy A.Regulation and misregulation of Eph/ephrin expression［J］.Cell Adh Migr, 2012,6(2):131-137.

［21］Fan R, Enkhjargal B, Camara R,et al. Critical role of EphA4 in early brain injury after subarachnoid hemorrhage in rat［J］.Exp Neurol, 2017,296:41-48.

［22］Lin JC, Lin SJ, Zhang JJ.Efficacy of tongxinluo capsules combined with nimodipine in patients with cerebral small vessel disease complicated with cognitive dysfunction［J］.Medical Innovation of China, 2022, 19(27): 45-49. (in Chinese)

林金财,林素桔,张佳佳.通心络胶囊联合尼莫地平对脑小血管病合并认知功能障碍患者的疗效［J］.中国医学创新,2022,19(27):45-49.

［23］Salvadori E, Poggesi A, Donnini I，et al.Efficacy and safety of the association of nimodipine and choline alphoscerate in the treatment of cognitive impairment in patients with cerebral small vessel disease. The CONIVaD trial［J］. Drugs Aging, 2021, 38(6):481-491.

[1]	吴俊波杨杰肖锋李金亮 . 微小RNA-138调控Wnt通路对体外人脑胶质瘤细胞生物学行为的影响 [J]. 解剖学报, 2023, 54(6): 682-688.
[2]	刘哲川李坤马帅男孟家琪王艳芹. 利拉鲁肽对百草枯诱导的小鼠帕金森病模型炎症及线粒体融合/分裂的影响 [J]. 解剖学报, 2023, 54(6): 676-681.
[3]	徐颖刘斌郑兴何宗钊. 补体C3a受体在盲肠结扎穿孔致脓毒症大鼠认知障碍中的作用及机制 [J]. 解剖学报, 2023, 54(6): 668-675.
[4]	许亚平余悦欣陈金福王柯柯郭志坤 . 高龄大鼠心室肌间质弹性纤维和胶原纤维的结构分布特征及其机制 [J]. 解剖学报, 2023, 54(6): 716-721.