电针重置超前性光暗周期转移小鼠的节律特征及其对SCN内相关钟基因表达的影响

发布时间：2018-01-28 09:25

本文关键词： 电针超前性光暗周期转移节律 SCN 钟基因　出处：《中山大学学报(自然科学版)》2017年02期 　论文类型：期刊论文

【摘要】：为了探索电针重置超前性光暗周期转移模型小鼠的时相特征及视交叉上核(Suprachiasmatic nucleus,SCN)多种节律相关基因及转录因子的表达,将符合筛选标准的44只C57BL/6J小鼠完全随机分为空白组(n=10)、模型组(n=12)、捆绑组(n=12)和电针组(n=10)4组。其中,模型组、捆绑组和电针组运用超前性光暗周期转移法进行造模,连续10 d。造模结束后,模型组小鼠于ZT18时相点处死取材;电针组小鼠恢复正常光暗交替(Light and Dark,LD)状态,并于ZT16时相点选取"肝俞"和"至阳"穴进行电针治疗,连续3次,每天1次;捆绑组采用与电针组相同的方法进行平行捆绑。捆绑及治疗结束后,处死动物并剥取SCN,采用PCR Array检测各组动物SCN内节律相关基因及部分节律相关转录因子相对表达量。实验结果显示:(1)电针重置昼夜节律结果:造模后,模型组、捆绑组及电针组与造模前比较峰相位、起始活动时间均显著超前,昼夜活动节律周期缩短(P0.05);再同步期,电针组峰相位、起始活动时间后移,与造模期及空白组比较具有统计学差异(P0.05);再同步第1天及第2天电针组小鼠的昼夜活动节律周期与空白组和造模前比较有统计学差异(P0.05),再同步第3天与空白组和造模前比较无统计学差异(P0.05)。(2)SCN节律相关基因及转录因子变化:与空白组比较,模型组小鼠的SCN节律相关基因表达上调8个(Aanat、Crx、Epo、Nkx2-5、Pax4、Prf1、Rora、Stat5a),下调4个(Egr1、Per1、Per3、Prokr2);与模型组比较,捆绑组节律相关基因上调3个(Esrra、Mat2a、Per3),下调11个(Cartpt、Crx、Epo、Kcnma1、Mtnr1b、Nkx2-5、Nms、Pax4、Prf1、Prkacb、Prkca);与模型组比较,电针组节律相关基因上调6个(Egr1、Esrra、Mat2a、Per1、Per3、Prokr2),下调21个(Aanat、Arntl、Cartpt、Crx、Csnk1e、Epo、Kcnma1、Mtnr1b、Myod1、Nkx2-5、Nms、Opn3、Pax4、Prf1、Prkacb、Prkca、Prkcb、Prokr2、Rora、Rorb、Slc9a3、Tgfb1)。以上实验结果提示电针可缩短超前性光暗周期转移小鼠的昼夜节律重置时间,加速紊乱昼夜节律的恢复,这可能是通过对SCN内Per1、Egr1、Aanat、Prokr2等节律相关基因的调控作用实现的。
[Abstract]:To explore the temporal characteristics and Suprachiasmatic nucleus of the suprachiasmatic nucleus of the supraoptic chiasma in an electroacupuncture resetting model mouse model of light and dark cycle transfer. 44 C57BL / 6J mice, which met the screening criteria, were randomly divided into two groups: control group (n = 10) and model group (n = 12). The model group, the binding group and the electroacupuncture group were established by the method of light and dark cycle transfer for 10 days. The mice in the model group were killed at the phase point of ZT18. The mice in the electroacupuncture group recovered to the normal light and dark alternating light and and LDD state, and selected "Ganshu" and "Zhiyang" acupoints at the ZT16 phase point for 3 consecutive times. Once a day; The animals in the binding group were subjected to parallel binding in the same way as those in the electroacupuncture group. After the binding and treatment, the animals were killed and the SCN was stripped. PCR Array was used to detect the relative expression of Rhythm related genes and some Rhythm related transcription factors in SCN of each group. The results showed that the electroacupuncture reset circadian rhythm: after modeling. In model group, binding group and electroacupuncture group, the peak phase was significantly earlier than that before model making, and the circadian cycle of activity was shortened (P 0.05). In resynchronization period, peak phase and initial activity time of electroacupuncture group moved backward, there was a statistical difference compared with model making period and blank group (P 0.05). The circadian activity rhythm of mice in the electroacupuncture group on day 1 and day 2 was significantly different from that in the blank group and before modeling (P0.05). On the 3rd day of resynchronization, there was no significant difference between the control group and the control group and before model making. There was no significant difference in the changes of the Rhythm related genes and transcription factors between the control group and the control group. The expression of SCN rhythm related genes in the model group was up-regulated. 4 Egr1, Per1, Per3, Prokr2a were down-regulated; Compared with the model group, the Rhythm related genes in the binding group were up-regulated by 3 Esrragnon Mat2aP3 and down regulated by 11 CartptTX CrxEpoxEpo-Kcnma1. Mtnr1b1, Nkx2-5, Pax4, Prf1, Prkacb1, Prkcaanus; Compared with the model group, the rhythm related genes of electroacupuncture group were up-regulated by 6 Egr1 Egr1, Esrrag, Mat2a, Per1, Per3, Prokr2, and down-regulated by 21 Aanat. Arntl-CartptCsnk1eCsnk1eCnma1Mtnr1b (Mtnr1b) Myod1Nkx2-5 (NMSS) Opn3Pax4. Prf1,Prkacb,Prkca,Prkcb,Prokr2,Rora,Rorb,Slc9a3. The above results suggest that electroacupuncture can shorten the time of circadian rhythm reset and accelerate the recovery of abnormal circadian rhythm in mice with light and dark cycle metastasis, which may be through the treatment of Per1 in SCN. The regulation of Rhythm related genes such as Egr1 Aanatron Prokr2 was realized.
【作者单位】：成都中医药大学;
【基金】：国家自然基金面上项目(81373739) 四川省教育厅项目(15ZB0089)
【分类号】：R245
【正文快照】： divided into 4 groups which were the blank group(n=10),the model group(n=12),the bindinggroup(n=12)and the electro-acupuncture group(n=10).We housed mice in LD cycle for 10 days.The control group was kept 12 h∶12 h LD of 20 days,and then SCN collected a

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