| Purpose: By observing the effect of electroacupuncture and liver Yu on the expression of liver clock gene Cry1 and Cry2 in mice with disturbed sleep-wake cycle,the peripheral mechanism of electroacupuncture on sleep-wake cycle disorders was explored.Material and method:SPF grade,male C57BL/6J mice,6 weeks old,a total of 72,72 mice were randomly divided into blank group,model group,liver group and non-acupoint group.After 1 week of adaptive domestication in an indoor SPF room at 20℃ to 25℃,a 2h/2h short-term deprivation method was used to establish a model for 5 days.The general state of the mice was monitored by video tracking daily,and daytime activity was observed.Rest,nocturnal sleep,change in sleep-wake time,increased frequency of activity during the day,emotional irritability,increased offensiveness,observation of general survival conditions and the blank group are significantly different,that the model standards,stop modeling and begin electroacupuncture treatment.The model group was only given the same daily fixation,without acupuncture intervention;electroacupuncture group to take bilateral "Gan Shu" points for needle acupuncture,inward and downward oblique 0.4cm,then 6805-AII type electroacupuncture instrument For treatment,both ends of the output lead were connected to the bilateral "Gan Shu" points.Electroacupuncture stimulation parameters: Dense wave 4/20Hz;Dense cycle of 6 seconds,the current was adjusted to the tail or the lower extremity slightly twitch.From 15:00 to 17:00 every day,each treatment for 15 minutes,a total of 5 days;non-acupoints selected mice fixed under double stress,acupuncture and electroacupuncture group.Detection behavior,analysis of data;mice were sacrificed at 02:00,06:00,10:00,14:00,16:00,20:00,6 time points(3 at each time point)for each group.The liver tissue was taken and the expression of Cry1 mRNA and Cry2 mRNA were detected by real-time fluorescence PCR.The expression of Cry1 and Cry2 protein was detected by Western blot.Results: 1.The number of standing times showed that compared with the blank group,the number of times of activity and standing times of the mice in the model group was statistically significant(P<0.05);the number of standing and activities of the liver group was decreased compared with the model group.(P<0.05).2.The weight results showed that there was no significant difference in body weight between the mice of each group before modeling(P>0.05).Before each group of mice was treated,the body weights of the mice in each group were different(P<0.05).The weight of the model group,Ganshu group and non-acupoint group was less than that of the blank group(all P<0.05).After treatment,compared with the blank group,the weight difference between the model group and the non-acupoint group was statistically significant(P<0.05);the weight of the mice in the Ganshu group was not statistically significant(P>0.05).3.The results of molecular level showed that the expression of Cry1 mRNA and Cry2 mRNA in mice blank group and Ganshu group showed obvious circadian rhythm(P<0.3).Compared with the blank group,the expression of Cry1 and Cry2 protein in the model group increased statistically(P<0.05).Compared with the model group,the expression of Cry1 and Cry2 protein in the Ganshu group was significantly lower(P<0.05).Compared with the liver Yu group,the non-acupoint group Cry1,Cry2 protein content was significantly higher(P <0.05).Conclusion: 1.Electroacupuncture liver Yu can restore the circadian rhythm of Cry1 mRNA and Cry2 mRNA in C57BL/6J mice with sleep-wake cycle,and it can regulate C57BL/6J mice with disturbed sleep-wake cycle.2.Sleep-wake cycle rhythm disorder C57BL/6J mice Cry1,Cry2 protein content increased,acupuncture liver Yu sleep-wake cycle disorder C57BL/6J mice Cry1,Cry2 protein content in the liver,thereby regulating sleep-Awakening cycle rhythm. |
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