Based On PI3K/AKT/mTOR Pathway To Explore The Mechanism Of Qingre Huazhuo Jiangsuan Decoction In Treating Acute Gouty Arthritis In Rats

Objective:To observe the effect of Qingre Huazhuo Jiangsuan Decoction on autophagy based on PI3K/AKT/mTOR signal pathway,evaluate the therapeutic effect of Qingre Huazhuo Jiangsuan Decoction on rats with acute gouty arthritis(AGA)by influencing autophagy at different doses,and explore the possible mechanism of Qingre Huazhuo Jiangsuan Decoction in the treatment of AGA,providing scientific basis for the clinical treatment of acute gouty arthritis in traditional Chinese medicine,and also providing new basis and ideas for the prevention and treatment of AGA by traditional Chinese medicine.Methods:88 SPF grade male SD rats were randomly divided into 8 groups after the end of adaptive feeding:blank group,model group,high dose group of traditional Chinese medicine,medium dose group of traditional Chinese medicine,low dose group of traditional Chinese medicine,colchicine group,rapamycin group and 3MA group.On the the fourth day of administration,the acute gouty arthritis model was established by injecting sodium urate(MSU)suspension into the knee joint.On the the fourth day of administration,the acute gouty arthritis model was established by injecting sodium urate(MSU)suspension into the knee joint for one week.After the last administration,the synovial tissue of the knee joint was taken out.The swelling degree of knee joint of rats in each group was measured within 48 hours after modeling,the pathological state of synovial tissue in each group was evaluated by hematoxylin-eosin(HE)staining,and TNF-α、IL-6 and IL-1β was detected by enzyme-linked immunosorbent assay(ELISA).The mRNA expression of NLRP3,ATG5,ATG7,PI3K,AKT and mTOR were detected by qRT-PCR.Western blot was used to detect the protein expression of LC3II/I,p62,ULK1,P-ULK1,Beclin-1,PI3K,AKT,mTOR,P-PI3K,P-AKT and P-mTOR.Results:Within 48 hours after modeling,the swelling degree of knee joint in the model group was significantly higher than that in the blank group,indicating that the modeling was successful.The swelling degree in the high,middle and low dose groups of traditional Chinese medicine and the colchicine group was significantly lower than that in the model group.The quantitative results of HE staining pathological sections showed that the pathological degree of the high,middle and low dose groups of traditional Chinese medicine and the colchicine group was significantly lower than that of the model group;The levels of TNF-α、IL-6、IL-1β、NLRP3 inflammatory factors in the high,medium,and low dose groups of traditional Chinese medicine and the colchicine group were significantly reduced compared to the model group;Compared with the model group,the protein(mRNA)expression of PI3K,AKT,mTOR,P-PI3K,P-AKT and P-mTOR in the high,middle and low dose groups of traditional Chinese medicine was significantly lower;Compared with the model group,the protein(mRNA)expression of LC3II/I,Beclin-1,ATG5 and ATG7 in the high,middle and low dose groups of traditional Chinese medicine was significantly higher,and the p62 and PULK1/ULK1 were significantly lower.Conclusions:Both Qingre huazhuo jiangsuan Decoction and colchicine can effectively alleviate the inflammatory reaction and the pathological degree of knee joint synovium in rats with acute gouty arthritis.Qingre huazhuo jiangsuan Decoction can effectively inhibit the PI3K/AKT/mTOR pathway and promote autophagy.Rapamycin can activate autophagy,and 3MA can inhibit autophagy.With the activation of autophagy,the pathological degree of knee joint can be effectively alleviated,It is suggested that Qingre huazhuo jiangsuan Decoction may alleviate acute gouty arthritis by inhibiting PI3K/AKT/mTOR pathway and promoting autophagy.