| Purpose:To observe the effect of acupuncture on the expression of melatonin in the liver and brain optic nerve cross nucleus of mice without circadian rhythm C57BL/6J,to study the rhythm of acupuncture on the content of melatonin in brain optic nerve cross nucleus and liver tissue,and to explore the mechanism of acupuncture on circadian rhythm disorder C57BL/6J sleep in mice.Methods: A 360 monitoring device was used to monitor and record the autonomous activity and standing of C57BL/6J mice.For C57BL/6J 144 mice,The 144 mice were randomly divided into blank group,model group,acupuncture group,non-menstrual group,36 in each group.Alternate acclimation according to light and dark(Light:08:00-20:00Dark:20:00-08:00),The mouse model without circadian rhythm was obtained by Roman Yasenkov short time deprivation protocol 2 h/2h light stimulation after one week of domestication.Among them,The blank group did not do any treatment during acupuncture;The model group only made the model without acupuncture(tied and fixed on the self-made mouse bundle during acupuncture intervention);The acupuncture group was treated with electroacupuncture for 15 minutes at 15:00,Current intensity m A,0.2 15 Hz,frequency For 5consecutive days,Acupuncture at bilateral Ganshu point in the direction of the lower oblique prick 0.4 cm,At non-acupoint site(double flank fixation),0.4 cm was obliquely spiked inward and downward.RT-q PCR and elisa were used to detect the expression of melatonin in brain and liver tissues of mice every two hours.Results:1.through the animal behavior trajectory analysis system of the scientific research platform,the variance analysis of the diurnal activity of mice was carried out.There was no significant difference(P>0.05)between the groups before modeling.There was significant difference between model group,acupuncture group and non-acupoint group(P<0.01),model group,acupuncture group and non-acupoint group(P>0.05)during the day after modeling.Night after modeling: model group,acupuncture Ganshu group and non-acupoint group and blank group(P<0.01)were statistically significant;model group,acupuncture Ganshu group and non-acupoint group(P>0.05)were not statistically significant.2.The changes of melatonin rhythm in 24 h brain and liver tissues of mice were calculated by linear regression equation of language software,The results showed that the expression of MTm RNA in the SCN of brain showed obvious changes of rhythmic oscillation in blank group(P=0.01<0.05),non-rhythmic oscillation in model group(P=0.74>0.05),rhythmic oscillation in gene expression recovery in Ganshu group(P=0.01),and non-rhythmic oscillation in non-cavity group(P=0.69>0.05);The expression of MT in liver tissue: the blank group showed obvious rhythmic oscillation(P=0.0005<0.05),the model group gene expression no rhythmic oscillation(P=0.85>0.05),the Ganshu group gene expression recovery rhythmic oscillation(P=0.00049<0.05),the non-pavoid group gene expression did not restore rhythmic oscillation(P=0.84>0.05).3.RT-qPCR detection of melatonin expression in the brain,The highest point in the blank group was between 22 and 2,Point 0 is its highest point,The lowest point is between 10 and14,The lowest point is 12;The highest point of the model group appears at 22,The lowest point appears at 4;The highest point of Ganshu group was between 22 and 2,Point 0 is its highest point,The lowest point is between 10 and 14,The lowest point is 12;The highest point in the non-acupoints group was 0,The lowest point appears at 2.Compared with the blank group(P<0.05),the acupuncture group was statistically significant;Comparison between model group and blank group(P>0.05)was not statistically significant;Compared with model group and non-menstrual and non-acupoint group(P>0.05),there was no statistical significance.4.Elisa detection of melatonin expression in liver,The highest point in the blank group appears between 12:00 and 16:00,14 is the highest point,The lowest point is between 2 and 6,Four points are the lowest point;The highest point of the model group and the non-non-point group appeared at 14,The lowest point of the model group appeared at 10 o'clock,and the lowest point of the non-menstrual group appeared at 18 o' clock;The highest point of Ganshu group was between 12:00 and 16:00,14 is the highest point,The lowest point is between 2 and 6,Four is the lowest point.Compared with the blank group(P<0.05),the acupuncture group was statistically significant;Comparison between model group and blank group(P>0.05)was not statistically significant;Compared with model group and non-menstrual and non-acupoint group(P>0.05),there was no statistical significance.Conclusion:The results of RT-qPCR and Elisa test showed that acupuncture of Ganshu could restore the rhythmic oscillation of melatonin in the brain and liver tissue of mice with circadian rhythm disorder.Rhythmic concussion of peripheral liver may affect the rhythmic concussion SCN central brain is one of the sleep regulation mechanisms in mice. |
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