| Objective: The aim of the this study was to investigate the effect of electroacupuncture intervention on circadian rhythm disorder mice,in terms of the gene expression and protein expression in C57BL/6J mice.Methods: The frequency of spontaneous activity and standing of the rhythm circadian disorder mice was monitored and recorded by 360°intelligent camera.This trial randomized mice into: blank group,model group,ganshu group,and sham electroacupuncture group in a1:1:1:1 ratio.Each group received LD(Light : 08:00-20:00,Dark : 20:00-08:00)domestication for 7days.From the eighth day,short-time sleep deprivation i.e.alternating2-hour darkness with 2-hour light was taken for 5 days.After those processes,from the 13 th day,the ganshu group and sham electroacupuncture group were bundled and administered on electroacupuncture treatment at Ganshu(BL18),the stimulation parameters are : frequency of2-15 Hz,current intensity of 0.2mA,session duration of 20 minutes once a day at 15:00 for 5consecutive days.The blank group and model group were bundled without electroacupuncture treatment.Samples were taken at 2 am on the 18 th day of the experiment per 4 hours,6 times in total,and 3 mice kidneys were taken from each group.Fluorescent quantitative PCR and Western Blot were used to detect the mRNA expression levels and protein expression levels of Clock and Bmal1 gene in mice kidney,and the law of cosines was used to prove the rhythm of gene expression.Results:1.The 2-hour sleep deprivation method was used to establish model in C57BL/6J mice.After that,there were significant changes in mood,activity increasing,standing frequency increasing,feed reducing,disturbance of activity initiation time,fur changing and defecation.In model group,ganshu group and sham electroacupuncture group,the frequency of spontaneous activities and standing were significantly increased compared with blank group(P < 0.05).2.After the intervention,the frequency of spontaneous activities and standing increased significantly in the model group compared with the ganshu group(P < 0.05).Compared withthe sham electroacupuncture group,the frequency of spontaneous activities and standing was decreased in the ganshu group which had statistical significance(P < 0.05),while there were no statistical difference(P>0.05)between sham electroacupuncture group and the model group.3.The mRNA expression of Bmal1 and Clock recovered in the ganshu group with statistical significance(P<0.3).The mRNA expression of Clock and Bmal1 in the model group was rhythm disorder without statistical significance(P>0.3).The mRNA gene expressions of Bmal1 and Clock in the kidney in the ganshu group showed circadian rhythm,and the difference was statistically significant(P<0.3).The mRNA gene expressions of Bmal1 and Clock in the kidney in the model group and the sham electroacupuncture group showed no obvious circadian rhythm(P>0.3).4.The protein expression quantity of Clock gene in ganshu group was decreased and narrow-er compared with blank group,sham electroacupuncture group and model group,which suggested statistical significance(P < 0.05);Compared with the ganshu group,the model group was brighter and wider,and the expression level was increased(P<0.05)which was statistically significant;Compared with sham electroacupuncture roup,the bands of ganshu group was slightly darker and thinner,and the expression level was decreased,showed statistical significance(P<0.05);Compared with the blank group,the expression in the ganshu group was decreased statistically(P<0.05).There were no statistical difference between model group and sham electroacupuncture group(P>0.05).Conclusion:1.Using 2-hour sleep deprivation model establishment in C57BL/6J mice can obtain rhythm circadian rhythm disorder mice.2.Electroacupuncture on ganshu can decrease the frequency of standing and activity of C57BL/6J mice with circadian rhythm disorder.3.ElEctroacupuncture liver Yu has a downward effect on Clock and Bmal 1 proteins in the kidneys of mice without circadian node.It may be the role of peripheral liver and kidney clock system genes in the regulation,thus restoring circadian rhythm in mice without circadian rhythm. |
