Hou Shuai, Zheng Shen, Wang Jianing, Chen Xiong, Zhong Zelan, Xue Hong, Chen Shasha
College of Acupuncture-Moxibustion and Tuina, Chengdu University of TCM, Chengdu 610072, Sichuan Province, China.
Zhongguo Zhen Jiu. 2018 Aug 12;38(8):864-70. doi: 10.13703/j.0255-2930.2018.08.017.
To observe the effects of electroacupuncture (EA) on the circadian rhythm and suprachiasmatic nucleus (SCN) epigenetic modification in mice with hepatocellular carcinoma (HCC), and to explore the epigenetics mechanism of EA on circadian rhythm in patients with HCC.
According to six zeitbeger time (ZT) of ZT0 (7:00), ZT4 (11:00), ZT8 (15:00), ZT12 (19:00), ZT16 (23:00) and ZT20 (3:00), a total of 108 eligible male C57BL/6J mice were divided into a blank group, a model group and an EA group at each ZT, 6 mice each group. Injection of H22 cancer cell suspension was used to establish the HCC model. After 11 days, EA (2 Hz/15 Hz, 0.2 mA) for 10 days was applied at "Ganshu" (BL 18) and "Zhiyang" (GV 9) in the EA group at each ZT, once a day, 15 min a time; the rats in the blank group and model group were treated with immobilization at the same time and under the same conditions. ClockLab (ACT-500) software was used to record the activity rhythm of mice. After 10 days intervention, MATLAB (R2007b) was used to export the circadian rhythm of mice, and the amplitude and peak phase of the mice were analyzed. The high-throughput epigenetics PCRarray array was applied to detect epigenetics-related gene expression in SCN.
(1) After modeling, compared with the blank group, the amplitude of activity was decreased and peak phase was delayed in the model group and EA group at each ZT (all <0.05), but the difference of rhythm parameters between the model group and EA group was not significant (all > 0.05). (2) After intervention, compared with the model group, the amplitude of activity in the EA group at ZT 8 was increased and peak phase was advanced (both <0.05); the difference of the activity amplitude and peak phase between the EA group and model group at ZT0, ZT4, ZT12, ZT16 and ZT20 was not significant (all >0.05); compared with the ZT0, ZT4, ZT12, ZT16 and ZT20, the amplitude of activity in the EA group at ZT 8 was increased and peak phase was advanced (all <0.05). (3) The results of epigenetic PCRarray array showed that after intervention at ZT 8, compared with the blank group, the expression of 48 epigenetic-related genes in SCN of HCC mice was up-regulated; compared with the model group, the relative expression of 49 epigenetic-related genes in the SCN was down-regulated in the EA group; there were 23 epigenetic-related genes differentially expressed among the three groups.
EA has benign regulation on circadian rhythm of HCC mice, and achieves the best efficacy at ZT 8. EA at ZT 8 could down-regulate the overexpression of epigenetic-related genes.
观察电针（EA）对肝癌（HCC）小鼠昼夜节律及视交叉上核（SCN）表观遗传修饰的影响，探讨EA对HCC患者昼夜节律的表观遗传学机制。
根据六个时间点（ZT），即ZT0（7:00）、ZT4（11:00）、ZT8（15:00）、ZT12（19:00）、ZT16（23:00）和ZT20（3:00），将108只符合条件的雄性C57BL/6J小鼠在每个ZT分为空白组、模型组和EA组，每组6只。注射H22癌细胞悬液建立HCC模型。11天后，在每个ZT对EA组的“肝俞”（BL 18）和“至阳”（GV 9）施加EA（2 Hz/15 Hz，0.2 mA）10天，每天一次，每次15分钟；空白组和模型组大鼠在相同时间和相同条件下进行固定处理。使用ClockLab（ACT - 500）软件记录小鼠的活动节律。干预10天后，使用MATLAB（R2007b）导出小鼠的昼夜节律，并分析小鼠的振幅和峰值相位。应用高通量表观遗传学PCRarray阵列检测SCN中表观遗传学相关基因的表达。
（1）建模后，与空白组相比，模型组和EA组在每个ZT的活动振幅均降低，峰值相位延迟（均<0.05），但模型组和EA组的节律参数差异不显著（均>0.05）。（2）干预后，与模型组相比，EA组在ZT 8时的活动振幅增加，峰值相位提前（均<0.05）；EA组与模型组在ZT0、ZT4、ZT12、ZT16和ZT20时的活动振幅和峰值相位差异不显著（均>0.05）；与ZT0、ZT4、ZT12、ZT16和ZT20相比，EA组在ZT 8时的活动振幅增加，峰值相位提前（均<0.05）。（3）表观遗传学PCRarray阵列结果显示，在ZT 8干预后，与空白组相比，HCC小鼠SCN中48个表观遗传学相关基因的表达上调；与模型组相比，EA组SCN中49个表观遗传学相关基因的相对表达下调；三组之间有23个表观遗传学相关基因差异表达。
EA对HCC小鼠的昼夜节律有良性调节作用，在ZT 8时疗效最佳。ZT 8时的EA可下调表观遗传学相关基因的过表达。
