电针长强穴对FMR1基因敲除小鼠海马区CypA-ERK1/2通路及其阻断效应的研究
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摘要
目的观察电针长强穴对FMR1基因敲除小鼠海马区CypA、ERK1/2及CREB蛋白表达的影响及对其阻断效应的研究。方法选取FMR1基因缺失且日龄为25日的KO小鼠,通过聚合酶链式反应鉴定小鼠基因型,选取基因型为纯合子型的小鼠入组,随机分为空白组、长强穴组、非经非穴组和阻断剂长强穴组,共24只。所有组别的小鼠均于每日固定时间段进行干预,1天1次,连续干预14天。干预前、后连续两天于固定时间内对各组实验小鼠进行自主活动行为学测试。采取免疫组化法及免疫印迹法分析海马区CypA、ERK1/2、CREB蛋白的表达情况。结果①自主活动行为学测试结果:干预后长强穴组自主活动及站立次数低于其他组别;干预后自主活动及站立次数低于干预前。②免疫组化法检测结果:长强穴组CypA、CREB蛋白的阳性平均光密度表达高于空白组、非经非穴组和阻断剂长强穴组,具有统计学意义(P<0.05);长强穴组ERK1/2蛋白的阳性平均光密度表达高于空白组及阻断剂长强穴组,具有统计学意义(P<0.05)。③免疫印迹法检测结果:长强穴组CypA、ERK1/2及CREB蛋白的表达高于空白组、非经非穴组和阻断剂长强穴组,其中ERK1/2及CREB蛋白具有统计学意义(P<0.05)。结论①电针长强穴对实验小鼠自主活动站立行为有影响。②电针长强穴能促进FMR1基因敲除小鼠海马区CypA、ERK1/2及CREB蛋白的表达,且MEK阻断剂PD98059对此有阻断效应。
引文
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[2] 吴强,韩平,王振宇,等. 针刺长强穴治疗精神发育迟滞患儿57例多中心临床研究报告[J].中华中医药杂志,2011,26(11):2668.
[3] 邹园华. 针刺长强穴配合感觉统合训练治疗精神发育迟滞20例[J]. 中国中医药现代远程教育,2016,14(1):98.
[4] 杨莹. 海马依赖性情节记忆调控的突触和神经环路机制研究[D]. 华中科技大学博士学位论文,2012.
[5] 李忠仁. 实验针灸学[M]. 北京:中国中医药出版社,2003.
[6] 诸葛启钏.小鼠脑立体定位图谱[M]. 北京:人民卫生出版社,2005.
[7] 姚春鹏.黄帝内经[M]. 北京:中华书局,2014.
[8] 林栋,卜婉萍,杨晓婷. 电针长强穴对FMR1基因敲除小鼠小脑CypA相关蛋白表达的影响及其阻断效应研究[J]. 康复学报,2017,27(5):29.
[9] Colgan J, Asmal M, Neagu M, et al. Cyclophilin A regulates TCR signal strength in CD4+ T cells via a proline-directed conformational switch in Itk[J]. Immunity,2004,21(2):189.
[10] Mallis R J, Brazin K N, Fulton D B, et al. Structural characterization of a proline-driven conformational switch within the Itk SH2 domain[J]. Nat Struct Biol,2002,9(12):900.
[11] 李文博,刘薇,陈璨,等.胞外亲环素A对炎症发生的影响及其抗体在抑制炎症反应中的作用[J].生物工程学报,2018,34(1):90.
[12] Jin Z G, Melaragno M G, Liao D F, et al. Cyclophilin A is a secreted growth factor induced by oxidative stress[J]. Circ Res,2000,87(9):789.
[13] Yurchenko V, Zybarth G, O'Connor M, et al. Active site residues of cyclophilin A are crucial for its signaling activity via CD147[J]. J Biol Chem,2002,277(25):22959.
[14] Seko Y, Fujimura T, Taka H, et al. Hypoxia followed by reoxygenation induces secretion of cyclophilin A from cultured rat cardiac myocytes[J]. Biochem Biophys Res Commun,2004,317(1):162.
[15] Feld M, Dimant B, Delorenzi A, et al. Phosphorylation of extra-nuclear ERK/MAPK is required for long-term memory consolidation in the crab Chasmagnathus[J]. Behav Brain Res,2005,158(2):251.
[16] Pappas B A, de la Torre J C, Davidson C M, et al. Chronic reduction of cerebral blood flow in the adult rat: late-emerging CA1 cell loss and memory dysfunction[J]. Brain Res,1996,708(1-2):50.
[17] Barco A, Marie H. Genetic approaches to investigate the role of CREB in neuronal plasticity and memory[J]. Mol Neurobiol,2011,44(3):330.
[18] 胡亚卓,吕佩源. MAPK/ERK信号转导通路与学习记忆[J]. 中国神经免疫学和神经病学杂志,2006,13(6):369.
[19] Jiang M D, Zheng S M, Xu H, et al. An experimental study of extracellular signal-regulated kinase and its interventional treatments in hepatic fibrosis[J]. Hepatobiliary Pancreat Dis Int,2008,7(1):51.
[20] 吴忱莼,李留霞,郭瑞霞,等. ERK1/2通路抑制剂PD98059影响卵巢癌细胞SKOV3增殖、侵袭能力的研究[J]. 中国妇幼保健,2015,30(2):281.