电针对原位肝癌小鼠转轮活动节律及视交叉上核Per基因表达的影响  

Electroacupuncture Intervention Regulates Circadian Rhythms by Down-regulating Per Gene Expression in Hypothalamic Suprachiasmatic Nucleus of Hepatocellular Carcinoma Mice

作　　者：侯帅[1] 郑申[1] 陈雄[1] 王佳宁[1] 钟泽兰 陈莎莎[1] 魏大能[1] 薛红[1] HOU Shuai;ZHENG Shen;CHEN Xiong;WANG Jia-ning;ZHONG Ze lan;CHEN Sha-sha;WEI Da-neng;XUE Hong(Chengdu University of Traditional Chinese Medicine,Chengdu 610072,China)

机构地区：[1]成都中医药大学,成都610072

出　　处：《针刺研究》2018年第10期632-639,共8页Acupuncture Research

基　　金：国家自然科学基金项目(No.81373739);四川省教育厅基金项目(No.15ZB 0089);成都中医药大学自然科学青年创新基金(No.ZRQN 1613)

摘　　要：目的:观察电针对原位肝癌移植模型小鼠转轮活动节律的调节作用及其对视交叉上核(SCN)中核心钟基因Period(Per)1、Per 2表达的影响,探讨电针对原发性肝癌患者昼夜节律调整的作用机制。方法:雄性C 57BL/6J小鼠按6个授时因子时间(ZT)随机分为ZT 0组、ZT 4组、ZT 8组、ZT 12组、ZT 16组和ZT 20组,每组各设空白组、模型组和电针组3个亚组,每个亚组6只。肝脏局部注射H 22癌细胞株建立原位肝癌移植模型。电针组在各时间点给予电针小鼠双侧"肝俞"和"至阳",每次15min,每日1次,治疗10d。全程采用ClockLab(ACT-500)软件记录小鼠活动节律,采用MATLAB(R 2007b)导出小鼠转轮活动节律图谱,分析小鼠活动起始时间、振幅、峰相位、周期。HE染色观察肝癌组织癌变情况。实时荧光定量PCR法检测SCN内钟基因Per 1mRNA、Per 2mRNA的相对表达量。结果:肝癌小鼠活动振幅较空白组下降(P<0.05),不同时间点电针均能影响肝癌小鼠转轮活动的振幅及峰相位,尤以ZT 8电针能显著升高肝癌小鼠降低的振幅,前移滞后的峰相位(P<0.05);不同时间点电针能够不同程度下调肝癌小鼠SCN内Per 1mRNA、Per 2mRNA的相对表达量,尤以ZT 8最为显著(P<0.05)。结论:电针能够对肝癌小鼠转轮活动节律产生良性调节,且以ZT 8最佳。这一作用可能是通过下调SCN内Per 1mRNA、Per 2mRNA的相对表达量实现。Objective To observe the effect of electroacupuncture（EA）on the rhythm of running-wheel activity of hepatocellular carcinoma（HCC）mice and the expression of Per 1 and Per 2（circadian rhythm genes）in the hypothalamic suprachiasmatic nucleus（SCN）,so as to investigate its mechanism underlying regulating circadian rhythm.Methods A total of 108 male C 57 BL/6 Jmice were randomly divided into control,HCC model and EA groups which were further assigned to six zeitbeger（environmental light-dark cycle）time（ZT）point（ZT 0,ZT 4,ZT 8,ZT 12,ZT 16 and ZT 20）subgroups.The HCC model was established by injection of H 22 cancer cell（abdominal 3 rd generation,10!L）suspension into the larger live lobe.Mice of the control group received saline injection of the liver lobe.EA（2 Hz/15 Hz,0.2 mA）was applied to bilateral＂Ganshu＂（BL 18）and＂Zhiyang＂（GV 9）for 15 min,once daily for 10 days.Mice of the control and model groups received the same binding-fixing to those of the EA group.Circadian running-wheel activity of 12 h∶12 hlight darkness（LD）cycle（activity onset and acrophase of actogram,amplitude or peak of periodogram）was recorded by using ClockLab（ACT-500）software and analyzed by MATLAB（R2007 b）before and after EA treatment.The pathological changes of liver cells were observed under light microscope after sectioning and H.E.staining.The expression levels of Per 1 mRNA and Per 2 mRNA in the liver tissues were determined by fluorogenic quantitative real time-PCR.Results（1）Following modeling,the amplitude of periodogram of running-wheel activity was significantly lowered at ZT 0,ZT 4,ZT 8,ZT 12,ZT 16,and ZT 20 relevant to the control group（P〈0.05）.After EA intervention,the amplitude of periodogram at ZT 8（15：00）was considerably increased relevant to the model group（P〈0.05）,and the acrophase at ZT 8 was remarkably advanced（P〈0.05）.No significant changes were found in the onset time and periods of periodogram at the 6 time-points after modeling an

关 键 词：电针 转轮活动节律 肝癌小鼠 PER 1 PER 2 视交叉上核 

分 类 号：R245.97[医药卫生—针灸推拿学]