主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2017, Vol. 40 ›› Issue (8): 641-645.doi: 10.3969/j.issn.1006-2157.2017.08.005

• 博士之光 • 上一篇    下一篇

不同光制下小鼠下丘脑视交叉上核Clock mRNA昼夜表达特点及温胆汤的干预作用

杨阳1, 张明泉1, 蒲晓田2, , 谭东宇1, 马淑然3#,   

  1. 1 河北中医学院基础医学院 河北 050200;
    2 淄博职业学院;
    3 北京中医药大学
  • 收稿日期:2017-03-03 出版日期:2017-08-10 发布日期:2017-08-10
  • 通讯作者: 马淑然,女,博士,教授,博士生导师,研究方向:天人相应理论和实验研究,E-mail:mashuran64@sina.com
  • 作者简介:杨阳,女,博士,副教授,硕士生导师
  • 基金资助:
    河北省教育厅高等学校科学技术研究项目(No.QN2015043),河北中医学院博士基金课题(No.BSZ2014003),国家重点基础研究发展计划“973”计划项目(No.2001CB505101)

Experimental study on the different day-night expression of hypothalamus SCN Clock mRNA in mice under different light systems and Wendan Tang’s intervention*

YANG Yang 1, ZHANG Mingquan1, PU Xiaotian2,TAN Dongyu1, MA Shuran3#   

  1. 1 School of Preclinical Medicine, Hebei University of Chinese Medicine, Hebei 050200, China;
    2 Zibo Vocational College, Shandong 255314, China;
    3 Beijing University of Chinese Medicine, Beijing 100029, China
  • Received:2017-03-03 Online:2017-08-10 Published:2017-08-10
  • Supported by:
    Science and Technology Research Project of College and University, funded by Hebei Province Department of Education(No.QN2015043), National Key Basic Research and Development Program (973) Program (No.2001CB505101)

摘要: 目的 观察在不同光制下,一天中4个时间点,血清褪黑素水平和下丘脑视交叉上核(SCN)Clock mRNA基因表达的特点,以及温胆汤的干预作用。方法 2~3月龄雄性KM小鼠160只,随机分为正常组(6点开灯照明,18点关灯黑暗)、模型组(24 h光照)、黑暗组(24 h黑暗)、温胆汤组和褪黑素组,每组32只。自然授时光照时间用ZT表示、黑暗组时间用CT表示,于第12周的最后一天,自然授时光照起点凌晨6点(ZT0/CT0)、12点(ZT6/CT6)、18点(ZT12/CT12)、24点(ZT18/CT18)4个时间点,分别于每组中随机选取8只,剖杀取材。采用酶联免疫吸附法(ELISA)检测血清褪黑素含量。采用聚合酶链式反应(PCR)法,检测小鼠下丘脑SCN Clock mRNA的表达。结果 血清褪黑素水平方面:正常组小鼠在ZT18含量最高(P<0.05);黑暗组小鼠在CT0低于CT18和CT12时含量(P<0.05);模型组小鼠4个时间点的含量并无差异(P>0.05),温胆汤组血清褪黑素的含量ZT0和ZT6均低于ZT18(均P<0.05);褪黑素组血清褪黑素含量ZT0、ZT6、ZT12均低于ZT18(均P<0.05)。小鼠下丘脑SCN Clock mRNA表达水平方面:正常组小鼠下丘脑SCN Clock mRNA表达水平ZT18低于ZT6(P<0.05);黑暗组小鼠4个时间点没有差别(P>0.05);模型组小鼠ZT0低于ZT6和ZT18(均P<0.05),ZT12低于ZT6和ZT18(均P<0.05);温胆汤组和褪黑素组小鼠均ZT18低于ZT6(均P<0.05)。结论 温胆汤能够在一定程度上纠正因持续光照所致的褪黑素含量下降和生物钟基因表达紊乱。

关键词: 温胆汤, 褪黑素, 生物钟, 基因, 光照, 天人相应, 小鼠

Abstract: Objective To observe the characteristics of the hypothalamus SCN Clock mRNA expression in mice under four timing points in a day and light condition and the effects of Wendan Tang (Gallbladder-Warming Decoction, WDT). Methods 160 male KM mice of 2~3 months were randomly divided into normal group (6:00—18:00 light), dark group (24 h dark), model group (24 h light),WDT group, and melatonin group(n=32 in each group). Zeitgeber time (ZT) and Circadian time (CT) were used to indicate natural light and dark timing points respectively At the last day of the 12th week,8 mice from each group were randomly selected and sacrificed at natural light timing points of 6 a.m. (ZT0 / CT0), 12 (ZT6 / CT6), 18 (ZT12 / CT12), and 24 (ZT18 / CT18). The concentration of melatonin in serum was measured by using enzyme-linked immunosorbent assay (ELISA). The expression of SCN Clock mRNA in mouse hypothalamus was detected by using polymerase chain reaction (PCR) method. Results Serum melatonin: The melatonin level at ZT18 of mice in the normal group was the highest (P<0.05) among all groups. Serum melatonin level in mice of the dark group at CT0 was lower than those at CT18 and CT12 (P<0.05). There were no differences in the melatonin levels between four timing points in mice of the model group (P>0.05). Serum melatonin levels in mice of the WDT group were lower at ZT0 and ZT6 than that at ZT18 (P<0.05) while these values at ZT0, ZT6 and ZT12 were lower than that at ZT18 in mice of the melatonin group (P<0.05). Expression of SCN Clock mRNA in mice hypothalamus: The expression of SCN Clock mRNA was lower at ZT18 than that at ZT6 (P<0.05). No differences were found at the four timing points in mice of the dark group (P>0.05). The expression levels in mice of the model group were lower at ZT0 and ZT12 compared with ZT18 (P<0.05); also lower at ZT0 and ZT12 compared with ZT 6 (P<0.05). The expression levels in mice of WDT and melatonin groups at ZT18 were lower than those at ZT6 (P<0.05). Conclusion WDT is rich in melatonin and could adjust biological clock rhythm disorder and reduction of melatonin due to continuous light.

Key words: wendan tang, melatonin, biological clock, gene, light, correspondence between man and universe, mouse

中图分类号: 

  • R285.5