| Objectives:To observe the role of TRPV1-NO in anti-inflammatory effects of moxibustion.Methods:50 healthy male KM mice were randomly divided into control group,model group,moxibustion group,capsicin group,and moxibustion+capsazepine group,10 mice in each;16 healthy male mice of wild type C57 were randomly divided into wildtype model group and wildtype moxibustion group,each group has 8 mice;14 healthy male mice of J003770TRPV1 were divided into two groups randomly:TRPVl-/-model group and TRPV1-/-moxibustion group,each group has seven mice.The mice,except the KM mice in control group,were intradermal injected with Complete Freund's Adjuvant to establish the adjuvant arthritis model in the first experiment day,after 24 hours,if the scores of arthritis grade was more than 4,then the AA model was builed successfully.After the successful molding,mice in moxibustion group,widetype moxibustion group and TRPV1-/-moxibustion group accepted moxibustion therapy in L5,L6 region of spine,20min a day,seven days in total.Mice in capsicin group were subcutaneous injected with O.lml/lOg capsicin in L5,L6 region of spine once a day,seven days in total.Moxibustion+capsazepine group mice got the administration by intraperitoneal injection of 0.1mg/10g capsazepine for pretreatment 15min before moxibustion therapy in L5,L6 region of spine,once a day,seven days in total.Attention should be paid,when given the moxibustion,to the distance between moxa stick and the skin which needed to maintain 12±2mm,so as to guarantee the temperature of mice skin to be 44±1 ?.During the treatment period,there was no treatment in control group and model group.The right hindfoot paw volumes of mice in each group were measured and recorded on Day l,Day 2,Day 4,Day 6 and Day 8 of the experiment.In the eighth day of experiment,blood were drew after the treatments,the level of NO.iNOS.eNOS?nNOS and IL-1??TNF-? in serum were detected.Results:1.Compared with the model group,paw volumes and serum IL-1??TNF-?level of mice in moxibustion group had significantly reduced(P<0.01).2.After the pretreatment of capsazepine,the moxibustion(moxibustion+capsazepine group)could still reduce the paw volumes and serum IL-1? TNF-? level of model mice,and had significant difference with the model group(P<0.01);but the reduction effect was not as good as the moxibustion group(P<0.01).3.Paw volumes and serum IL-1??TNF-?level of mice in TRPV1-/-moxibustion group didn't markedly changed when compared with the TRPV1-/-model group(P>0.05).4.In the moxibustion group,the serum NO?iNOS and eNOS level were reduced markedly when compared with the model group(P<0.01),but the nNOS level didn't change much(P>0.05),and the correlation analysis showed that the serum's NO level was correlated with the paw volumes and serum IL-1??TNF-? level(P<0.05?P<0.05?P<0.01).5.After the pretreatment of capsazepine,the moxibustion(moxibustion+capsazepine group)could still reduce the serum NO?iNOS and eNOS level of model mice,and had significant difference with the model group(P<0.01);but the reduction effect was not as good as the moxibustion group(P<0.01).6.The serum NO?iNOS and nNOS level of mice in TRPV1-/-moxibustion group didn't markedly changed when compared with the TRPV1-/-model group(P>0.05),but the eNOS level reduced a lot(P<0.01).7.Using the partial correlation analysis method to analyze the correlation of NO with iNOS?eNOS in serum of mice in three moxibustion groups:moxibustion group,wildtype moxibustion group and TRPV1-/-moxibustion group,the results showed that partial correlation coefficient of NO with iNOS in three groups respectively were r=0.924?r=0.945 and r=0.946,partial correlation coefficient of NO with eNOS in three groups respectively were r=0.609?r=0.592 and r=0.433,indicated that the production of NO has a closer correlation with iNOS.Conclusions:TRPV1 plays an important role in the anti-inflammation effect of moxibustion,which may related to the participation of TPRV1 in the regulation of NO level during inflammation. |
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