Experimental Study of Huangqin Qingre Chubi Capsule on Regulating Synovial Cell Autophagy Through PI3K/Akt/mTOR Signaling Pathway for Improving Inflammation in Acute Gouty Arthritis Rats

ZHU Nanfei; CAO Yunxiang; LIU Jian; WANG Hui; WANG Amin

Abstract：Objective To investigate the effect of Huangqin Qingre Chubi Capsule(HQC)modulating synoviocyte autophagy on inflammation in rats with acute gouty arthritis through PI3K/Akt/mTOR signaling pathway.Methods A total of 72 male SD rats were randomly divided into normal group,model group,celecoxib group and HQC low,medium and high dose groups,with 12 rats in each group.The corresponding drugs were pre-administered for 5 days.The rat model of acute gouty arthritis was prepared by Coderre method 1h after administration on the 5th day.After modeling,the intervention groups continued to receive the drugs,during which the ankle swelling of the rats was measured.Half of the rats in each group were killed on the 7th and 14th day of administration,and the levels of TNF-α and IL-1β were detected by ELISA.HE staining was used to observe the pathological morphology of ankle synovial tissue.The expression levels of PI3K,Akt,mTOR,and the levels of phosphorylated proteins p-PI3K,p-Akt and p-mTOR,as well as the levels of NOD-like receptor protein 3(NLRP3)and microtubule-associated protein 1a/1b-light chain(LC3)in synovium were detected by Western blot.Results Compared with the normal group at the same time,the joint of rats in the model group was swollen and with a large number of inflammatory cell infiltration,the ratios of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR were significantly increased,while the expression level of LC3 protein was significantly decreased,as well as the levels of NLRP3 protein,TNF-α and IL-1β were significantly increased on the 7th and 14th day(P<0.01).Compared with the model group at the same time,the p-PI3K/PI3K,p-mTOR/mTOR ratio,TNF-α and IL-1β levels of HQC groups and celeoxib group were significantly decreased on the 7th day(P<0.01).The p-Akt/Akt ratio and NLRP3 protein expression level were significantly decreased in the HQC medium,high-dose groups,and celeoxib group(P<0.01),while LC3 protein expression level of HQC medium dose group was increased(P<0.05).On the 14th day,the p-PI3K/PI3K,p-Akt/Akt ratio,NLRP3 protein expression level,TNF-α and IL-1β levels in HQC groups and celeoxib group were significantly decreased(P<0.01),while the LC3 protein expression level was significantly increased(P<0.01).The p-mTOR/mTOR ratio significantly decreased in both the HQC medium and high dose groups,as well as in the celecoxib group(P<0.01).Compared with day 7,LC3 protein expression levels in HQC high dose group and celeoxib group were significantly increased on day 14,while NLRP3 protein expression levels,TNF-α and IL-1β levels in HQC groups and celeoxib group were significantly decreased(P<0.01).Conclusion HQC can improve acute gouty arthritis by inhibiting PI3K/Akt/mTOR signaling pathway,promoting autophagy of rat synoviocytes,inhibiting the activation of NLRP3 inflammasome.