基于网络药理学探讨恒古骨伤愈合剂治疗骨关节炎的机制及动物实验初步验证

Mechanism of Osteoking Improving Osteoarthritis based on Network Pharmacology and DDM Rats

目的探讨恒古骨伤愈合剂(OK)治疗骨关节炎(OA)的潜在机制并利用DMM大鼠初步验证。方法使用TCMSP数据库收集OK潜在的靶点,运用GeneCards数据库和OMIM筛选与OA相关的靶基因,利用Cytoscape3.8.2软件建立OK有效成分与OA靶基因间互作的调控网络。使用STRING数据库构建蛋白互作图,进行基因本体(GO)及京都基因与基因组百科全书(KEGG)分析。此外,采用内侧半月板不稳定术(DMM)构建OA大鼠,观察膝关节软骨病变,ELISA检测OK治疗后炎症以及氧化应激水平。结果网络药理学发现,血清白蛋白(ALB),白介素6(IL-6)等80个OK治疗OA的核心基因,主要涉及免疫、炎症反应、氧化应激等生物学过程。动物实验发现,与模型组相比,OK组大鼠关节软骨表面结构趋于正常,软骨细胞脱落少;OK降低血清炎症因子IL-6(P<0.01)、VEGF(P<0.01)和氧化应激因子MDA(P<0.05),改善ALB水平和SOD活性(P<0.01)。结论OK可缓解OA软骨组织损伤,改善免疫,抑制炎症和氧化应激反应,初步验证了网络药理学的结果。

Objective To explore the potential mechanism of Osteoking(OK)in the treatment of Osteoarthritis(OA)and preliminarily verify it with DMM rats.Methods TCMSP database was used to collect potential targets of OK,GeneCards database and OMIM were used to screen target genes related to OA,and Cytoscape 3.8.2 software was used to establish a regulatory network for interaction between OK Active ingredient and OA target genes.The STRING database was used to construct protein interaction maps and GO and KECG were analyzed.In addition,OA rats were constructed using medial meniscus instability surgery(DMM)to observe knee joint cartilage lesions,and ELISA was used to detect inflammation and oxidative stress levels after OK treatment.Results Network pharmacology results showed that 80 core genes were involved in OK therapy for OA,including serum albumin(ALB)and interleukin-6(IL6),mainly involving biological processes such as immunity,inflammatory response,oxidative stress,and metabolism.Animal experiments showed that compared with the model group,the surface structure of the articular cartilage in the OK group of rats tended to be normal,with less detachment of chondrocytes;OK reduced serum inflammatory factors IL-6(P<0.01),VEGF(P<0.01),and oxidative stress factor MDA(P<0.05),improved ALB levels and SOD activity(P<0.01).Conclusion OK can alleviate OA cartilage tissue damage,enhance immunity,and inhibit inflammation and oxidative stress reactions,preliminarily verifying the results of network pharmacology.