基于蛋白芯片技术探讨针刺对脑缺血大鼠脑组织相关磷酸化蛋白的影响  

Effect of Acupuncture on Neurological Function and Related Differentially-depressed Profiles in the Brain of Middle Cerebral Artery Occlusion Rats Using Antibody Chips Technique

作　　者：田浩梅[1] 颜虹[1] 张娟 贺平[1] 陈楚淘[3] TIAN Hao-mei YAN Hong ZHANG Juan HE Ping CHEN Chu-tao(Key Lab of Correlation between Meridian Acupoints and Viscera of Hunan Administration of Traditional Chinese Medicine, Hunan University of Traditional Chinese Medicine, Changsha 410208, China Ningxiang County Hospital of Traditional Chinese Medicine, Changsha 410600 2 Key Lab of Internal Medicine Constructed by Chinese Ministry of Education and Provincial Government, Hunan University of Traditional Chinese Medicine, Changsha 41000)

机构地区：[1]湖南中医药大学,湖南中医药管理局经穴与脏腑相关重点研究室,长沙410208 [2]湖南省宁乡县中医院,长沙410600 [3]湖南中医药大学省部共建中医内科学教育部重点实验室,长沙410007

出　　处：《针刺研究》2016年第6期479-485,共7页Acupuncture Research

基　　金：国家自然科学基金项目(No.81303051;No.30901901);中医内科学省部共建教育部重点实验室资助(No.ZYNK 201501)

摘　　要：目的:基于蛋白芯片技术探讨针刺对大脑中动脉阻塞(MCAO)大鼠脑组织相关磷酸化蛋白的影响,部分阐释针刺促脑组织修复的机制。方法:SD大鼠随机分为假手术组、模型组、对照点组、穴位组,10只/组。参照Longa线栓法并加以改良复制MCAO模型。穴位组与对照点组取"大椎""百会"和"水沟"穴或非穴对照点(穴位左侧旁开约3mm处的非经非穴点),每12h针刺1次,每次30min,治疗6次。实验结束后行神经功能缺损评分;采用720磷酸化抗体蛋白芯片技术检测各组大鼠脑组织损伤修复相关信号蛋白磷酸化的变化,筛选出各组差异性表达的蛋白。结果:与假手术组比较,模型组神经功能缺损评分显著升高(P<0.01);与模型组比较,穴位组与对照点组神经功能缺损评分降低(P<0.01,P<0.05),且穴位组低于对照点组(P<0.05)。蛋白芯片显示:与假手术组比较,模型组中蛋白的磷酸化水平发生了变化,以上调蛋白磷酸化水平为主,且其上、下调蛋白功能主要涉及细胞凋亡、细胞周期调节、细胞的分化与增殖、细胞骨架与连接、细胞信号转导、DNA的修复及转录等;与模型组比较,穴位组、对照点组多种蛋白的磷酸化水平均发生改变,穴位组蛋白磷酸化水平改变以下调为主,其主要功能类型与模型组发生改变的蛋白相似,而对照点组蛋白磷酸化水平的改变没有明显趋向,且发生改变的磷酸化蛋白数量与穴位组存在差异。结论:针刺"大椎""百会""水沟"穴可以通过多系统、多途径调整脑组织相关蛋白的磷酸化水平,从而促进脑组织损伤的修复。Objective To observe the effect of acupuncture on changes of neurological function and expression of proteins in the ischemic brain region in middle cerebral artery occlusion（MCAO）rats with protein chip technique,so as to reveal the profiles of cerebral proteins related to its effectiveness in improving cerebral ischemia（CI）.Methods SD rats were randomized into sham operation（sham）,CI model,non-acupoint and acupoint groups（n=10in each group）.The CI model was established by MCAO according to modified Longa＇s method.For rats of the acupoint group,＂Dazhui＂（CV 14）,＂Baihui＂（GV 20）and＂Shuigou＂（GV 26）were punctured by twirling the filiform needles for about 1min,and repeated once again during 30 min of needle retention,which was conducted once every 12 h,and 6times altogether.The non-acupoints were about 3mm away from the above-mentioned acupoints and stimulated with the same method and same procedures.The neurological deficit score was scaled according to Longa＇s method.The differentially-expressed proteins（≥1.5folds in up-and down-regulation）in the ischemic region of the brain were detected by using Springbio 720 antibody chip technology.Results Compared with the sham group,the neurological deficit score was significantly higher in the model group（P〈0.01）,while compared with the model group,the neurological deficit scores were considerably in both acupoint and non-acupoint groups（P〈0.05,P〈0.01）.The therapeutic effect of acupoint group was evidently superior to that of the non-acupoint group in improving neurological function（P〈0.05）.Compared with the sham group,the differentially-expressed proteins in the ischemic brain including 33up-regulated and 12down-regulated were found in the model group,mainly functioning in cell apoptosis,cell cycle regulation,cell differentiation and proliferation,cell cytoskeleton and connection,cell signal transduction,DNA repair and transcription factors.Compared with the model group,the differentially-expressed protein

关 键 词：脑缺血 手针 神经功能 大脑差异表达蛋白 抗体芯片 

分 类 号：R245.9[医药卫生—针灸推拿学]