目的 探讨电针治疗阿尔茨海默病的效果和作用机制。
方法 雄性8月龄APP/PS1小鼠24只随机分为模型组、电针组和非穴组,各8只。8只野生型小鼠为野生组。电针组电针百会、神庭,非穴组电针双侧肋下非穴区,野生组和模型组予相同抓取和固定,共28 d。干预后,Morris水迷宫测试小鼠学习记忆能力,分别采用免疫荧光和免疫组化技术检测小鼠大脑皮质β淀粉样蛋白(Aβ)和β位淀粉样前体蛋白裂解酶1 (BACE1)含量,RT-PCR检测BACE1 mRNA水平。
结果 与模型组比较,电针组小鼠Morris水迷宫逃避潜伏期显著下降(P < 0.001),穿越平台次数显著增加(P < 0.001),BACE1表达和Aβ水平显著减少(P < 0.001)。
结论 电针百会、神庭可能通过抑制APP/PS1小鼠大脑皮质BACE1表达,降低Aβ水平,改善小鼠学习记忆能力。
Objective To explore the effects of electroacupuncture at Baihui (DU20) and Shenting (GV24) on Alzheimer's disease, and possible mechanism for it.
Methods A total of 24 eight-month-old APP/PS1 male mice were randomly divided into model group (n = 8), electroacupuncture group (n = 8) and non-acupoint group (n = 8), and other eight wild-type mice were as wild-type group. The electroacupuncture group accepted electroacupuncture at Baihui and Shenting, while the non-acupoint group accepted electroacupuncture at bilateral subcostal non-acupoint area, and the wild-type group and the model group accepted the same grasping and fixing, for 28 days. Then they assessed with Morris water maze test. The levels of β-amyloid protein (Aβ) and β-site amyloid precursor protein cleaving enzyme 1 (BACE1) in cerebral cortex were detected with immunohistochemistry and immunofluorescence respectively, and the level of BACE1 mRNA with RT-PCR.
Results Compared with the model group, the escape latency decreased in the electroacupuncture group (P < 0.001), and the times crossing platforms increased (P < 0.001), while the expression of BACE1 and Aβ decreased (P < 0.001).
Conclusion Electroacupuncture may improve the learning-memory ability by inhibiting the expression of BACE1 in the cerebral cortex of APP/PS1 mice to decreasing the level of Aβ.
[1] Kumar A, Singh A, Ekavali. A review on Alzheimer's disease pathophysiology and its management: an update[J]. Pharmacol Rep, 2015, 67(2): 195-203.
[2] Barage S H, Sonawane K D.Amyloid cascade hypothesis: pathogenesis and therapeutic strategies in Alzheimer's disease[J]. Neuropeptides, 2015, 52(8): 1-18.
[3] Mullard A.Alzheimer amyloid hypothesis lives on[J]. Nature Rev Drug Discov, 2016, 16(1): 3-5.
[4] 曹江鹏,张利达,蔡兴慧,等. 近5年针刺改善学习记忆临床应用与作用机制研究概况[J]. 辽宁中医药大学学报, 2017, 19(05): 122-124.
[5] Perng C H, Chang Y C, Tzang R F.The treatment of cognitive dysfunction in dementia: a multiple treatments meta-analysis[J]. Psychopharmacology, 2018, 235(5): 1571-1580.
[6] Liu W, Zhuo P, Li L, et al.Activation of brain glucose metabolism ameliorating cognitive impairment in APP/PS1 transgenic mice by electroacupuncture[J]. Free Radic Biol Med, 2017, 112(11): 174-190.
[7] 陈吉祥,吴羽楠,郑雅媗,等. 电针百会穴对APP/PS1双转基因小鼠学习记忆及脑源性神经营养因子表达的影响[J]. 中国康复理论与实践, 2015, 21(6): 642-647.
[8] 李忠仁. 实验针灸学[M]. 北京:中国中医药出版社, 2003: 325-329.
[9] Braak H, Braak E, Bohl J.Staging of Alzheimer-related cortical destruction[J]. Eur Neurol, 1993, 33(6): 403-408.
[10] Delacourte A, David J P, Sergeant N, et al.The biochemical pathway of neurofibrillary degeneration in aging and Alzheimer's disease[J]. Neurology, 1999, 54(52): 1158-1165.
[11] 陈玉鹏,林丹红,陈立典,等. 中医认知功能理论体系的构建[J]. 中医杂志, 2016, 57(1): 12-15.
[12] Wang F, Zhong H, Li X, et al.Electroacupuncture attenuates reference memory impairment associated with astrocytic NDRG2 suppression in APP/PS1 transgenic mice[J]. Mol Neurobiol, 2014, 50(2): 305-313.
[13] Li G, Zhang X, Cheng H, et al.Acupuncture improves cognitive deficits and increases neuron density of the hippocampus in middle-aged SAMP8 mice[J]. Acupunct Med, 2012, 30(4): 339-345.
[14] Bilkeigorzo A.Genetic mouse models of brain ageing and Alzheimer's disease[J]. Pharmacol Therapeutics, 2014, 142(2): 244-257.
[15] Lee J E, Han P L.An update of animal models of Alzheimer disease with a reevaluation of plaque depositions[J]. Exp Neurobiol, 2013, 22(2): 84-95.
[16] Garciaalloza M, Robbins E M, Zhangnunes S X, et al.Characterization of amyloid deposition in the APPswe/PS1dE9 mouse model of Alzheimer disease[J]. Neurobiol Dis, 2006, 24(3): 516-524.
[17] 赵凌,张富文,张虹,等. 电针治疗轻度认知功能障碍:多中心随机对照研究[J]. 中国针灸, 2012, 32(9): 779-784.
[18] 吴羽楠,陈吉祥,陶静,等. 电针百会对APP/PS1转基因小鼠学习记忆能力及Tau蛋白磷酸化的影响[J]. 中国康复医学杂志, 2015, 30(5): 432-436.
[19] Bodendorf U, Danner S, Fischer F, et al.Expression of human beta-secretase in the mouse brain increases the steady-state level of beta-amyloid[J]. J Neurochem, 2010, 80(5): 799-806.
[20] Sathya M, Premkumar P, Karthick C, et al.BACE1 in Alzheimer's disease[J]. Clinica Chimica Acta, 2012, 414(4546): 171-178.
[21] Vassar R.BACE1 inhibitor drugs in clinical trials for Alzheimer’s disease[J]. Alzheimers Res Ther, 2014, 6(9): 89-103.
[22] Yan R.Stepping closer to treating Alzheimer’s disease patients with BACE1 inhibitor drugs[J]. Transl Neurodegener, 2016, 5(1): 13-24.
[23] Dong W G, Wang F, Chen Y, et al.Electroacupuncture reduces Aβ production and BACE1 expression in SAMP8 mice[J]. Front Aging Neurosci, 2015, 7(7): 148-155.
[24] Lan Z, Cong S, Jin C, et al.Icariin decreases the expression of APP and BACE-1 and reduces the β-amyloid burden in an APP transgenic mouse model of Alzheimer's disease[J]. Int J Biol Sci, 2014, 10(2): 181-191.
[25] Tamayev R, Matsuda S, Arancio O, et al.β-but not γ-secretase proteolysis of APP causes synaptic and memory deficits in a mouse model of dementia[J]. EMBO Mol Med, 2012, 4(3): 171-179.
[26] Kim S, Sato Y, Mohan P S, et al.Evidence that the RAB5 effector APPL1 mediates APP-βCTF-induced dysfunction of endosomes in Down syndrome and Alzheimer's disease[J]. Mol Psychiatry, 2016, 21(5): 707-716.
[27] Lauritzen I, Pardossi-Piquard R, Bourgeois A, et al.Intraneuronal aggregation of the β-CTF fragment of APP (C99) induces Aβ-independent lysosomal-autophagic pathology[J]. Acta Neuropathol, 2016, 132(2): 257-276.
