Study On Anti-osteoarthritis Effects Of Ruyi Zhenbao Pills And Its Mechanism

OBJECTIVE : To explore the anti-osteoarthritis effect and pathological mechanism of Ruyi Zhenbao Pills(RZP),a Chinese Tibetan medicine,the models of inflammation and osteoarthritis(OA)in vivo and in vitro were constructed to investigate the protective effect and potential mechanism of RZP based on the relevant network pharmacological and molecular docking data.The results further clarify the clinical indications of RZP,which is conducive to the development and application of ethnic medicine prescription preparations.METHODS :(1)Combined with Traditional Chinese Medicine Systems Pharmacology,Pub Med platform,Traditional Chinese Medicine Integrated Database and literature,the chemical components of 30 medicinal materials in RZP were counted and their active components were screened.Swiss target prediction,OMIM,gene cards and PHARMGKB databases were used to search for OA related protein targets,and the potential disease targets of RZP according to active components.Analysis software Cytoscape 3.9.0 was used to obtain the intersection target of the two,and enrich and analyze the signal pathway of RZP in the treatment of OA,thus preliminarily predicting the mechanism of RZP in the treatment of OA.(2)The inflammatory model of ATDC5 cells induced by lipopolysaccharide was established in vitro,and the anti-inflammatory targets and pathways of RZP were evaluated and verified based on the network prediction results.The rat OA model induced by papain was constructed,and the pathological changes such as oxidative stress and inflammatory reaction in the knee joint of the OA rats were detected under the intervention of RZP,so as to explore the therapeutic effect of RZP on cartilage matrix and subchondral bone.RESULTS:(1)The results of network pharmacology showed that RZP contained556 active chemical components and 1602 disease targets.There are 110 OA targets,including 31 intersection targets.After further analysis,36 core components of RZP were screened,and the core targets include IL-6,TNF-α,IL-1β,COX-2,CXCL-8,NOS2,MMP-1,MMP-3,etc.The results of KEGG and GO enrichment analysis showed that TNF signaling pathway,NF-kappa B signaling pathway,MAPK signaling pathway,rheumatoid arthritis and osteoclast differentiation may be the key pathways of RZP in the treatment of OA.Molecular docking results show that β-sitosterol has the best binding activity with NOS2 and can be used as a key target for RZP in the treatment of OA.(2)Experiments in vitro showed that RZP had protective effects on the inflammatory model of ATDC5 cells induced by lipopolysaccharide,and inhibited inflammation by down regulating the relative m RNA expression of proinflammatory factors as TNF-α,IL-1β,COX-2 and i NOS.The results of animal experiments showed that RZP can effectively improve the joint swelling and threshold of heat and mechanical pain in OA rats induced by papain.And RZP effectively inhibited the expression of MMP-1 in serum,reduced the content of MDA in serum,increased the content of SOD,and reduced the relative m RNA and protein expression of proinflammatory factors including NO,TNF-α,IL-1β,COX-2 and i NOS in knee tissue,up regulated the expression of type II collagen and down regulated the expression of osteopontin in knee tissue,which restored the morphological changes of cartilage matrix and subchondral bone in rats to varying degrees.CONCLUSION: RZP can effectively alleviate the pain and inflammatory symptoms of OA via multiple targets,reduce the pathological changes of knee joint,which provides a theoretical basis for the rational treatment of OA in clinic.