Effects Of Electro-acupuncture At Different Time Points On Circadian Rhythm Of Liver Cancer Mice And It's Study Of The Effect On Per Gene

Objective:the aim of this study was to investigate the circadian rhythm characteristics of liver cancer mice in LD patients,and the effect of electro-acupuncture to adjust the role of its pathological rhythm,and to preliminary explore the mechanism of the correlation of Per gene involved in electro-acupuncture on the pathological rhythm of HCC mice.Methods:(1)The circadian rhythms of locomotor activity of the HCC mice were monitored and recorded by Data Acquisition and Processing System for locomotor activity rhythm for Small Animals and ClockLab Running Wheel Data Acquisition System.126 animals which were screened out were divided randomly into 6 time points'groups:the blank group,the model group and the model group(6 time points:ZT0,ZT4,ZT8,ZT12,ZT16,ZT20),7 mice per group.Each group received LD(LD12:12,7:00~19:00 for the light period,19:00~7:00 for the night period)domestication for 10 days before model establishment started.According to the reference,(2)The model group and the acupuncture group to the replicated hepatocarcinoma mouse mode with the 0.01 mL of H22 hepatocarcinoma cell suspension was injected along the long axis of the mouse liver,and the blank group was injected with the same dose of saline.(3)After the successful model establishment,the circadian rhythms of locomotor activity of mice were monitored for another 10 days.On the 11~thh day,start electroacupuncture intervention.Electro-acupuncture was administered at Zhiyang(GV 9)and Ganshu(BL 18)with HANS Acupoint and Nerves Stimulator(HANS-200)using 2/15Hz and 0.5mA for 15 min each time and one time a day for 10 days.Blank group and model group are not acupuncture,but taking the same stimulus bundle.The mice were sacrificed after 2h of the 10th electroacupuncture,and remove the material to be tested(4)Samples were then extracted and preserved for indexes testing with 6samples for RT-PCR testing in each group.The levels of Per1mRNA,Per2mRNA in SCN.The effect of electroacupuncture at 6 different time points on circadian rhythm of HCC mice and the effect of related Per gene were studied by statistic analysis of rhythm parameters before and after modeling and electroacupuncture.Results:(1)Before model eastablishment,there were no significant differences in the periods,start time,amplitudes,phase,phase positions and total activities of locomotor activity rhythms of the mice in blank and model groups(p>0.05).(2)After model eastablishment,the amplitudes and total activities of the mice in model group were decreased and the phase delayed.There were significant differences in the differences of amplitudes,phase positions and total activities before and after model eastablishment between blank group and model group(p<0.05),but there were no significant differences in the periods and medians of locomotor activity rhythms of the mice in the two groups(p>0.05).(3)After electro-acupuncture,the amplitudes were increased and the phases were lagged in the electro-acupuncture group.There were significant differences in the differences of amplitudes and phase positions before and after electro-acupuncture between model group and electro-acupuncture group(p<0.05)but there were no significant differences in the differences of the periods,medians and total activities before and after electro-acupuncture between model group and electro-acupuncture group(p>0.05).The electro-acupuncture effect was most obvious especially when electro-acupuncture at ZT8.(4)After model eastablishment,there were up in the expressions of the core per1 and Per2 clock genes in SCN of the mice in model group.the expressions compared with blank group were significant(p<0.05).In the ZT0,ZT4,ZT8,ZT20 phase of the electro-acupuncture can be a certain degree of down-regulation of SC1 Per1 gene expression level,and compared with the model group were statistically significant(P>0.05);In the ZT4,ZT8,ZT12 phase of the electro-acupuncture can be a certain degree of down-regulation of SC2 Per2 gene expression level,and compared with the model group were statistically significant(P>0.05).However,both ZT8 electroacupuncture at the time point can significantly down-regulate the expression of the above two genes in SCN.Conclusion:(1)After model establishment,the circadian rhythm of liver cancer mice became abnormal,electroacupuncture can restore its abnormal rhythm on a certain extent,however,in the ZT8 phase to adjust the rhythm of the electro-acupuncture effect is most significant.(2)The relative expression of Per1 and Per2 genes in SCN of HCC mice was abnormally increased,electroacupuncture can down-regulated and the relative expression of the two genes in some extent,and the most significant at ZT8.(3)The effect of electroacupuncture on Per1 and Per2rhythmic genes in SCN of abnormal HCC mice may be the molecular mechanism of electroacupuncture adjustment of circadian rhythm in SCN.