Effects Of Juanbi Lijieqing Recipe On Inflammatory Factors And NLRP3 Inflammasome In Rats With Acute Gouty Arthriti

Objective : 1.To study the anti-inflammatory effect of Juanbilijieqing Recipe on rats with acute gouty arthritis.2.To explore the possible relationship between Juanbilijieqing formula and NLRP3 inflammatory body in exerting anti-inflammatory mechanism.Methods : Sixty male SD rats of SPF grade were weighed and numbered.According to the body weight number,12 rats were randomly selected as the blank group(A)by random number table.Forty-eight rats were modeled with hyperuricemia,and then the gouty arthritis model was prepared.After successful modeling,the rats were randomly divided into model group(B),Juanbi Lijie Qingfang group(C),Etocoxi group(D),and positive drug isoliquiritigenin group(E),with 12 rats in each group.Four hours after modeling,two rats from each group were randomly selected and sacrificed,and the synovial tissue of knee joint was taken for pathological observation.After successful modeling,model group(B)and blank control group(A)were given the same amount of normal saline by gavage,and each group was given corresponding drug intervention.The gait and knee circumference of rats were observed before and 4h,12 h,24h,48 h after modeling.After 48 h of intervention,blood samples from abdominal aorta were collected to detect serum uric acid level in rats;hematoxylin-eosin(HE)staining was performed to observe the pathological changes of soft tissue around the knee joint in rats under a microscope;the concentrations of IL-1β and TNF-α in serum of rats and the contents of IL-1β,NLRP3,ASC and caspase-1 proteins in synovial tissues were detected by enzyme-linked immunosorbent assay(ELISA).Results :(1)Compared with the blank group(A),the rats in the modeling groups(B,C,D,E)were mentally apathetic and had poor gait,with grade 3 gait and obvious joint swelling(P < 0.05).Compared with the model group(B),the gait scores and joint circumference changes of rats in Juanbi Lijie Qingfang group(C),etoricoxib group(D)and isoliquiritigenin group(E)were improved to varying degrees at 24 h and 48 h after drug intervention(P < 0.05).(2)The tissue around the knee joint of rats was observed by light microscope.In the blank group(A),only a small amount of inflammatory cell infiltration and cell edema were observed in the synovial tissue of rats.The distribution of synovial cells was uniform and the synovial epithelium was complete.Compared with the blank group,the synovial tissue of the model group(B)rats had severe inflammation,the synovial epithelium was missing,accompanied by obvious cell edema and a large number of inflammatory cells such as neutrophils,and a small amount of sodium urate crystals.Compared with model group(B),Juanbi Lijie Qingfang group(C),Etoricoxib group(D)and Isoliquiritigenin group(E)showed similar performance.The cells in synovial tissue returned to normal,some synovial epithelium was intact,inflammatory cell infiltration was less,and a small part of cell edema was visible.(3)Compared with the blank group(A): After 48 hours of drug intervention,the levels of serum uric acid,IL-1β and TNF-α in the model group(B)were significantly increased(P <0.01);compared with model group(B),the levels of serum uric acid,IL-1β and TNF-α in Juanbi Lijieqing prescription group(C),etoricoxib group(D)and isoliquiritigenin group(E)were significantly decreased(P < 0.05).(4)Compared with the blank group(A): After 48 hours of drug intervention,the protein levels of NLRP3,ASC,caspase-1 and IL-1β in the synovial tissue of the model group(B)were significantly increased(P < 0.01);compared with model group(B),the protein levels of IL-1β,NLRP3,ASC and caspase-1 in synovial tissue of rats in Juanbi Lijieqing prescription group(C),etoricoxib group(D)and isoliquiritigenin group(E)decreased(P< 0.05).Conclusion:(1)Intraperitoneal injection of uric acid oxidase inhibitor potassium oxazinate combined with intra-articular injection of sodium urate can successfully replicate the rat model of hyperuricemia and acute gouty arthritis.(2)Juanbilijieqing recipe can improve the gait,joint swelling and other general conditions of rats with hyperuricemia and acute gouty arthritis,reduce the level of serum uric acid and reduce IL-1 β、TNF-α It can inhibit the infiltration of inflammatory cytokines in the joint cavity and improve the inflammation of synovial tissue in rats,indicating that juanbilijieqing formula has the inhibitory effect on gouty arthritis factors.(3)Juanbilijieqing recipe can inhibit the activation of NLRP3 inflammatory bodies,block the complex inflammatory cascade,protect synovial endothelial cells and reduce IL-1 β The differentiation and maturation of other inflammatory factors can achieve the mechanism of anti gout inflammation.