The Response Of Hypothalamic Pain-related Signaling Molecules In Moxibustion Analgesia

Objective : To study the influence of moxibustion on the analgesic effect of inflammatory pain and the expression of pain-related molecules in the hypothalamus.Methods:A total of 48 healthy male C57 BL / 6J mice(clean,6-8W,weight 20±2g,the baseline of each group is 13s-18s)were selected as the study subjects,The mice were randomly divided into control group,CFA model group,CFA model +moxibustion group(referred to as CFA+moxi group).CFA model group and CFA+moxi group were administered right foot plantation complete Freund’s adjuvant on the preparation for inflammatory pain models.CFA+moxi group started the moxibustion intervention on the 4th day after the model was established.The right Zusanli points(ST 36)was selected and the warm moxibustion was continued for 30 minutes.The other groups were crawled in the same way and intensity.The latency of thermal stimulation(Thermal withdrawal latency,TWL)was measured before and after modeling,before moxibustion,after moxibustion 0 minutes,30 minutes,60 minutes,90 minutes,120 minutes to determine the mouse thermal pain threshold.The hypothalamus of mice was removed immediately after the end of pain threshold determination,and the expression of hypothalamic pain-related molecules was verified by high throughput PCR assay and Q-PCR.Results:(1)Heat pain threshold results:Comparison among groups: Before modeling,analysis of three groups in mice thermal pain threshold(Baseline)(P> 0.05).After modeling,the thermal pain threshold of model group and moxibustion group decreased,Compared with the control group(P <0.01).It was no difference in model group and moxibustion group(P> 0.05).Group comparison: Compared with its own before modeling,TWL of model and the moxibustion group was declining obviously(P <0.01).Compared with modeling after treatment 0 minutes,30 minutes,60 minutes,90 minutes,120 minutes the heat pain threshold was increased significantly in moxibustion group(P <0.01).(2)The expression of pain-related molecules in the hypothalamus on moxibustion analgesia by PCR Array:Compared with the control group,there were 7 pain-related genes of hypothalamus significantly expressed in the model group,the up-regulation expression was 4(up-regulation more than 2 fold).CCL12,CCR2,IL-1α,TAC1 gene.There were three genes down-regulated,Ntrk1 gene,Scn10 a and Slc6a2 genes.Compared with the model group,13 pain-related molecules were significantly expressed in the moxibustion group,and all were down-regulated genes,CCL12,CCR2,IL-1α,CD4,CX3 cr,EDNRA,IL-1β,IL-6,PTGS2,TLR4,CNR1,SCN10 a,TRPa1 gene.(3)Q-PCR technique to verify hypothalamic pain-related molecular results:Compared the results of Q-PCR and PCR array between groups,it showed that CCL12 and CCR2 were up-regulated respectively after modeling,compared with the control group;The chemokines CCL12 and CCR2 were down respectively after moxibustion,compared with the model group.It can be observed that the results trends of Q-PCR are consistent with the PCR array between groups,suggesting that the result of PCR array is accurate and credible.Conclusion:(1)Moxibustion Zusanli on chronic inflammatory pain model mice can have a good analgesic effect;(2)Hypothalamus involved in moxibustion analgesia;(3)The effect of moxibustion analgesic may be related to the regulation of CCL12,CCR2,CX3CR1,IL-1α,IL-1β,PTGS2 and other genes in the hypothalamus.