基于腺苷A3受体途径针刺对足三里浅筋膜cAMP/Epac及Ras/MAPK途径的调节作用研究

Regulatory effect of acupunctue on cAMP/Epac and Ras/MAPK pathway in Zusanli superficial fascia based on adenosine A3 receptor pathway

目的:探讨针刺对足三里浅筋膜cAMP/Epac、Ras/MAPK途径的调节及其与腺苷A3受体途径之间的关系。方法:大鼠随机分为对照组、针刺组、针刺+IB-MECA组和针刺+Reversine组,各组在针刺后20min取足三里穴位区浅筋膜标本,HE和Masson染色检测针刺对组织结构及胶原沉积的影响,Real-Time PCR检测cAMP、Epac、Ras、ERK1/2 mRNA表达,Western Blot检测cAMP、Epac、Ras、p-ERK1/2、ERK、p-P38、P38等蛋白表达。结果:针刺可引起浅筋膜组织细胞排列紊乱,有细胞脱落现象,cAMP、Epac mRNA表达下调,Ras、ERK1/2 mRNA表达上调,cAMP、Epac信号蛋白表达下调,Ras、p-ERK1/2、p-P38信号蛋白表达上调,腺苷A3受体激动剂IBMECA可加强上述改变,而腺苷A3受体拮抗剂Reversine可减弱该效应。结论:腺苷A3受体激活后镇痛可能与上调浅筋膜组织Ras/MAPK途径、下调CAMP/Epac途径有关。

Objective:To investigate the regulation of acupuncture on cAMP/Epac,Ras/MAPK pathway and the relationship between acupuncture and adenosine A3 receptor pathway.Methods:The rats were randomly divided into control group,acupuncture group,acupuncture+IB-MECA group and acupuncture+Reversine group.The superficial fascia samples of Zusanli(ST 36)area were taken 20 minutes after acupuncture in each group.HE staining and Masson trichrome staining kit were used to detect the effects of acupuncture on tissue structure and collagen deposition.Real-Time PCR was used to detect the expression of camp,Epac,RAS and ERK1/2 mRNA.Western Blot was used to detect camp,Epac,RAS,p-ERK1/2,ERK1/2,p-P38 and P38.Results:Acupuncture can cause the disorder of cell arrangement and cell detachment in the superficial fascia.The expression of camp and Epac mRNA is down regulated,the expression of Ras and ERK mRNA is up regulated,the expression of camp and Epac signal protein is down regulated,the expression of RAS,p-ERK1/2 and p-P38 signal protein is up regulated.IBMECA,the activator of adenosine A3 receptor,can enhance the above changes,while Reversine,the antagonist of adenosine A3 receptor,can weaken the effect.Conclusion:The activation of adenosine A3 receptor may be related to the up regulation of Ras/MAPK pathway and down regulation of cAMP/Epac.