DNA Methylation In Anti-inflammatory Effect Of Moxibustion

Objective:(1)To study the effects of DNA methylation/methylolation balance of mice skin and the expression of epigenetic modifications related genes in different brain regions by long-term moxibustion;(2)To study DNA methylation modification on anti-inflammatory effect of moxibustion.Methods:First,the clean grade adult C57BL/6J mice,weight 20±2g,as our experimental object,were randomly divided into non moxibustion group,7days moxibustion group,15 days moxibustion group,30 days moxibustion group.Mice on the moxibustion group were taken moxibustion at Zusanli 30 min every day,and mice on the non moxibustion group were bound at the same way 30 min.The mice were killed at the end of the last intervention,moxibustion on the side Zusanli skin(local skin),moxibustion on the same side hip skin(distal skin),moxibustion against the side Zusanli skin(contralateral skin),the DNA methylation and methylation rate of the skins were detected by 5mC ELISA and 5hmc ELISA method.Then we used Epigenetic PCRarray to screen differentially expressed genes in hippocampus,hypothalamus,olfactory bulb and sensory cortex of mice on moxibustion for 30 days and moxibustion for 30 days.Second,the C57BL/6J mice were randomly divided into control group,model group(and moxibustion group after basal volume filter.Mice on the induced inflammatory model group were injected Freund's adjuvant into plam.On the 4th day after modeling,mice on the moxibustion group were taken moxibustion at Zusanli 30 minutes.Mice on the control group and the model group were taken the same way 30 minutes.The foot volume was measured every 4 days during the intervention period.Serum,moxibustion Zusanli,moxibustion ipsilateral hip skin(distal skin)and moxibustion on Zusanli skin side(contralateral skin)of mice were treated with enzymes after being killed at the end of the last intervention.The serum IL-1?,5mc and 5hmc detected by enzyme-linked immunosorbent assay.The data wasanalysed by SPSS 21.0 statistical software and graphs were plotted by GraphPad Prism 7.Results:1.Effects of DNA methylation and methylolation rate of mice skin.There was no significant difference in DNA methylation rate and methylolation rate between local skin,distal skin and contralateral skin in non-moxibustion group(P>0.05).Compared with non-moxibustion group,The rate of local skin DNA methylation and methylolation increased significantly in 30 days moxibustion group.(P<0.01).2.Expression of DNA methylation related genes in different brain regions of mice by 30 days moxibustionCompared with the non-moxibustion group,the expression of Dnmt3 a in the olfactory bulb was down-regulated by 13.64-fold in the 30-day moxibustion,2.38-fold in the hypothalamus,2.39-fold in the hippocampus,and 2.25-fold in the sensory cortex.3.Anti-inflammatory effects of moxibustion on chronic inflammation miceCompared with the control group,the foot volume was significantly increased in the model group and the moxibustion group(P<0.01),and the serum IL-1? was significantly increased(P<0.01);compared with the model group,the Foot volume in the moxibustion group began to decrease on the 19 th day of intervention.(P<0.05).Foot volume was significantly reduced from the 24 th day of intervention to the end of the final intervention(P<0.01),and serum IL-1? was significantly decreased after the last intervention(P<0.05).4.Effect of moxibustion on DNA methylation and methylolation rate of chronic inflammation mouse skinCompared with the control group,the DNA methylation rate of the contralateral skin and the distal skin in the model group was significantly decreased(P<0.05),and the DNA methylolation rate was significantly increased(P<0.05);In comparison,the DNA methylation rate of the skin,distal skin and contralateral skin in the moxibustion group was significantly higher(P<0.05),and the DNA methylolation rate wassignificantly decreased(P<0.05);Compared with local skin,the DNA methylation rate of the distal skin and the contralateral skin in the moxibustion group significantly increased(P<0.05),and the DNA methylation rate decreased significantly(P<0.05).Conclusions:1.Long-term moxibustion could regulate the balance of mice skin DNA methylation/methylolation modification at acupoints;2.Moxibustion had effects on the expression of DNA methylation-related genes in different brain regions and mice in 30 days,mainly concentrated on Dnmt3 a and Dnmt3 b,and both were down-regulated,.3.Long-term moxibustion could adjust the mice systemic skin DNA methylation/methylolation imbalance caused by chronic inflammation.