Inhibitory Effect Of Pulsatilla Saponins On Acute Gouty Arthritis In Rats Via NALP3 Inflammatory Body Signaling Pathway

Objective: To investigate the changes of NALP3 signaling pathway by observing the degree of joint swelling,serum inflammatory factors IL-1?,IL-8 and TNF-?in rats with acute gouty arthritis,to evaluate the therapeutic effect of Pulsatilla saponins and to analyze the related mechanisms,and to explore the efficacy and safety of Pulsatilla saponins for acute gouty arthritis,so as to provide experimental basis for the development of new clinical drugs.Methods: Forty male Wistar rats were randomly divided into blank group,model group,colchicine group and Pulsatilla saponin group after routine feeding for one week.Each group was administered continuously for one week.After measuring the ankle circumference of rats on the fourth day of administration,all the rats except blank group were injected with sodium urate solution into the ankle joint cavity to make the model.After the model was established,the rats were given intragastric administration for 3 days and measuring the ankle circumference of rats at each time point.(The circumference of ankle joint was measured at 6h,12 h,24h,48 h and 72 h after establishing the model.)The blood from inner canthus of rats was taken at 24 h and 72 h after model establishment and serum was prepared.The concentration of IL-1?,IL-8 and TNF-?was measured by ELISA;The synovial tissue was taken to observe the infiltration of synovial inflammatory cells by HE staining;The expression of NALP3 inflammatory body was observed by immunohistochemical staining;The expression of NF-kappa B and NALP3 in synovial tissue was measured by Western Blot method.Result: 1.Changes of ankle swelling in rats of each group At 6 hours after modeling,there were differences between colchicine group and Pulsatilla saponins group(P < 0.05),while there was no difference in ankle swelling between colchicine group and Pulsatilla saponins group(P > 0.05);there was no significant difference in ankle swelling between blank group and model group at 48 and 72 hours after modeling,and there was no difference in ankle swelling between model group and Pulsatilla saponins group at 6 and 12 hours after modeling(P > 0.05).At other time points,there was significant difference in the degree of ankle swelling between groups(P < 0.05).2.The expression of IL-1?,IL-8 and TNF-?in serum of rats in each group At 24 hours and 72 hours after the establishment of the model,the levels of IL-1?,IL-8 and TNF-?in the serum of rats in each group were significantly different(P < 0.05),and the blank group < the Pulsatilla saponins group <the colchicine group<the blank group.3.Histopathological changes in rats Pathological sections of model group showed a large number of inflammatory cell infiltration,colchicine group and Pulsatilla saponin group decreased inflammatory cell infiltration,Pulsatilla saponin group decreased more significantly.4.Expression of NALP3 and NF-kappa B in rat synovium The expression of NALP3 and NF-kappa B was significantly increased in the model group by the method of immunohistochemistry and Western blot.The expression of NALP3 and NF-kappa B was decreased in colchicine group and Pulsatilla saponin group,while the expression of NALP3 and NF-kappa B was least in Pulsatilla saponin group.Conclusion: 1.Pulsatilla saponins can regulate the swelling degree of ankle joint and improve the joint symptoms of gouty arthritis rats.2.Pulsatilla saponins can reduce the levels of serum inflammatory factors IL-1?,IL-8 and TNF-?.3.Pulsatilla saponins can inhibit the expression of NALP3 and NF-kappa B in synovial tissues,and inhibit the onset of acute gouty arthritis in rats by blocking the NALP3 inflammatory body signaling pathway.