补肾壮膝方治疗膝骨关节炎的作用机制研究OA

[Objective]To explore the potential molecular mechanism of Bushen Zhuangxi Formula(BSZXF)in the treatment of knee osteoarthritis(KOA)with kidney-yang deficiency based on ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS)technology,network pharmacology and animal experiments.[Methods]UPLC-Q-TOF-MS was used to qualitatively analyze the chemical composition of BSZXF.Swiss Target Prediction and other databases were used to obtain the component targets,and the intersections with KOA disease targets were taken to construct the protein-protein interaction(PPI)network,and GO and KEGG enrichment analysis were performed.The KOA rat model of kidney-yang deficiency was established by ovariectomy combined with intra-articular injection.The rats were randomly divided into sham operation group,model group,and BSZXF low,medium,and high-dose groups.After 28 days of administration,the syndrome of kidney-yang deficiency,pain threshold and knee joint swelling degree were observed.ELISA was used to detect related indicators in serum and synovial fluid.The qRT-PCR was used to detect the expression of key genes in PI3K-AKT and NF-κB pathways in articular cartilage tissue.[Results]Seventy-four compounds were identified from BSZXF.PPI network showed that TNF,IL6 and AKT1 were the key targets.KEGG enrichment analysis showed that PI3K-AKT and NF-κB signaling pathways were the core pathways.Animal experiments have shown that BSZXF can significantly improve the syndrome of kidney-yang deficiency,increase the pain threshold,and reduce cartilage damage.qRT-PCR results showed that compared with the blank group,the mRNA expressions of PI3K,AKT,NF-κB p65,and IκBα in cartilage tissue of the model group were significantly increased(P<0.05 or P<0.001).Compared with the model group,the high-dose BSZXF group significantly down-regulated the mRNA expression of all the above genes(P<0.05 or P<0.001).[Conclusion]BSZXF can improve the symptoms of KOA rats with kidney-yang deficiency and delay cartilage degeneration.The mechanism may be related to the crosstalk between PI3K-AKT and NF-κB pathways,which reflects the characteristics of multi-components,multi-targets and multi-pathways of traditional Chinese medicine.