The Influence Of Moxibustion With Different Temperature On Local Skin Mitochondrial Respiratory Chain And SIRT1,SIRT3

ObjectiveTo observe the influence of mitochondrial respiratory chain complex I~V and SIRT1,SIRT3 in local skin tissue caused by different temperature of moxibustion.To partly investigate the local skin mechanism of energy metabolism administrated by moxibustion.MethodsMice were randomly divided into control group,38°C moxibustion(38°C-moxi)and 46°C moxibustion(46°C-moxi)groups.Mice received moxibustion at Zusanli(ST36)acupoint for 30 min once.The local skin tissue of mice in each group were respectively taken at two time point(30 min and 72 h)after moxibustion.Temperature was controlled by a digital thermodetector.The enzymatic activities of mitochondrial respiratory chain(MRC)complexes I ~ V were assayed spectrophotometrically.Immunofluorescence(IF)and Western Blot(WB)were used to detect the protein expression of SIRT1 and SIRT3 in the skin.Results1 The enzymatic activities of MRC complexes I~V on local skin after completion of moxibustion: at 30 min after moxibustion,the enzymatic activities of MRC complexes I~V of 38°C and 46°C moxibustion groups were obviously higher than the control group(P<0.05).Compared with 38°C moxibustion group,the enzymatic activities of MRC complexes I~V were decreased significantly in 46°C moxibustion group(P<0.05).At 72 h after moxibustion,in contrast to control group,mice subjected to moxibustion at 38°C demonstrated increased the enzymatic activities of MRC complexes I~V.However,there was no statistical significance difference between the control group and 38°C moxibustion group.In addition,compared with 38°C moxibustion group,the enzymatic activities of MRC complexes I~V were decreased slightly in 46°C moxibustion group(P>0.05).These results were suggested that the enzymatic activities of MRC complexes I~V increased significantly after moxibustion(moxi-38°C > 46°C).Moreover,enzymatic activities of MRC complexes I~V arrived their peaks at 30 min after moxibustion and returned to normal level 72 h after moxibustion in 38°C moxibustion group.2 The protein expression of SIRT1 and SIRT3 on local skin after completion of moxibustion was evaluated by IF and WB.At 30 min after moxibustion,Compared with the control group,the protein expression of SIRT1 and SIRT3 in moxibustion groups were obviously increased(P<0.05).Compared with 38°C moxibustion group,the protein of SIRT1 and SIRT3 were increased significantly in 46°C moxibustion group(P<0.05).At 72 h after moxibustion,in contrast to control group,mice subjected to moxibustion at 38°C demonstrated increased the protein of SIRT1 and SIRT3.However,there was no statistical significance difference between the control group and 38°C moxibustion group.In addition,compared with 38°C moxibustion group,the protein expression of SIRT1 and SIRT3 were increased in 46°C moxibustion group(P<0.05).These results were suggested that the protein expression of SIRT1 and SIRT3 increased significantly after moxibustion(moxi-46°C >38°C).Moreover,the protein expression of SIRT1 and SIRT3 at 72 h after moxibustion continue to increase.However,there was no statistical significance difference between 30 min and 72 h after of moxibustion.Conclusions1 The enzymatic activities of mitochondrial respiratory chain and SIRT family protein on local skin participated in the initial process of energy metabolism or energy conversion of moxibustion.2 Moxibustion with different temperature had different effect on the specific mechanism of energy conversion.Moxibustion at 38°C triggered increased significantly of MRC complexes I~V expression.However,moxibustion at 46°C triggered increased significantly the protein expression of SIRT1 and SIRT3.3 Moxibustion triggered more significan increased the enzymatic activities of MRC complexes I~V after 30 min intervention.However,moxibustion triggered more significant increase the protein expression of SIRT1 and SIRT3 at 72 h after moxibustion.