Mechanism Of Anti-inflammation And Analgesia Of N~6-(2-Hydroxyethyl) Adenosine On Gouty Rats

Gout,an extremely painful arthritis with relapsing inflammatory attacks,is initially triggered by deposition of monosodium urate(MSU)crystals in the joints.N~6-(2-Hydroxyethyl)adenosine(HEA),one of main bioactive components in the medicinal mushroom Ophiocordyceps sobolifera,had been proved to possess anti-inflammatory and analgesic properties.In current study,transcriptome sequence was performed to understand global effects of HEA on gouty rats.Sequence results showed that 932 genes were differential expressed in HEA-induced gouty rats.Comparison of these 932 genes with the pain gene database,where the genes were changed during pain,57 genes were supposed to be highly potential to participate in the regulation of pain.According to related reports,12 anti-nocieptive genes were obtained including up-regulated adenosine A1 receptor(A1R,Adora1)and A2A receptor(A2AR,Adora2a).The results of western blot showed that A1R and A2AR,the regulated target of pain,were markedly increased and decreased respectively in HEA group(7.5mg/kg)as compared with model group,and the effect of HEA on A1R was reversed by the administration of selective A1R antagonist DPCPX.Sequence results also suggested that inflammatory response and autophagy may be altered by HEA.Hematoxylin-eosin(HE)staining showed HEA restrained inflammatory infiltration in ankle.HEA was able to decrease the swelling rate of ankle.The results of ELISA showed that interleukin 1?(IL-1?)and tumor necrosis factors?(TNF-?)were negatively regulated by HEA in gouty rats.Western blot results showed that HEA attenuated inflammation by suppressing NLRP3inflammasome activation through restoration of autophagic flux,which was dependent upon the activation of adenosine A1 receptor.These results demonstrated that HEA possess remarkable anti-inflammation and anti-nocieptive effect on gouty rats,and HEA exert its anti-inflammation effects though A1R-dependent restoration of autophagy as well as promoted degradation of p62 to suppresse NLRP3inflammasome activation and thus decreased the level of IL-1?and TNF-?.HEA may exert its antinociception by the activation of A1R,down-regulated A2AR and regulation of other pain-related genes.