龟板抑制破骨分化改善骨质疏松性骨折

Plastrum testudinis relieves osteoporotic fracture via inhibiting osteoclast differentiation

目的基于网络药理学和实验验证探讨龟板(plastrum testudinis,PT)治疗骨质疏松性骨折(osteoporotic fracture,OF)的潜在机制。方法借助BATMAN数据库得到PT成分及其对应靶标,检索GeneCards、OMIM数据库获得OF相关靶点,在STRING数据库输入交集靶点得到蛋白互作(protein-protein interaction,PPI)信息,借助Cytoscape3.7.2软件进行PPI网络及“龟板-活性成分-靶标-骨质疏松性骨折”网络的构建,使用Cytoscape3.7.2软件、R软件进行GO功能富集分析及KEGG通路富集分析。此外,为了验证网络药理学研究结果的可靠性,还进行体内实验验证龟板改善OF的作用,并通过体外实验探究了龟板干预下的人外周血单核细胞(human peripheral blood monocytes,HPBMs)表型。结果获得龟板6个活性成分,342个作用靶标,其中与OF相关靶标34个,GO功能富集分析共有802个结果(P<0.05),KEGG富集分析有67个结果,其中相关信号通路有22条(P<0.05),主要涉及TNF、MAPK、Estrogen、NF-κB等信号通路。值得一提的是,体内实验证实龟板可有效治疗OF模型大鼠的骨缺损;体外实验发现龟板能抑制RANKL诱导的HPBMs破骨分化。结论龟板可能通过多种化合物、靶点和通路调节炎症反应、激素代谢及细胞周期来治疗OF,其中抑制破骨分化可能是龟板抗骨质疏松性骨折的重要机制。

Objective To investigate the underlying mechanism of plastrum testudinis(PT)in the treatment of osteoporotic fracture(OF)using network pharmacology and experimental validation.Methods The components of PT and their corresponding targets were obtained with the help of BATMAN database,and relevant targets of OF were obtained by searching Genecards and OMIM databases.Intersection targets were input into the STRING database to obtain protein-protein interaction(PPI)information.The constructions of PPI network and PT-compound-target-OF network were done with Cytoscape 3.7.2,and GO function enrichment and KEGG pathway enrichment analyses were finished with Cytoscape3.7.2 and R.Additionally,to verify the reliability of the network pharmacology findings,we also conducted in vivo experiments to verify the effect of PT on improving OF and in vitro experiments,which investigated the phenotypes of human peripheral blood monocytes(HPBMs)under the intervention of PT.Results Six active compounds and 342 targets of PT were obtained,including 34 targets correlated with OF.There were 802 results gained through GO function enrichment analysis(P<0.05),and 67 results through KEGG pathway enrichment analysis,including 22 relevant signaling pathways(P<0.05),which were mainly involved in TNF,MARK,estrogen,NF-κB,and other signaling pathways.Notably,our in vivo experiments proved that PT could effectively treat the bone defects in OF model rats.The in vitro experiments showed that PT could inhibit RANKL-induced osteoclast differentiation in HPBMs.Conclusion By regulating inflammatory response,hormone metabolism,and cell cycle,PT may treat OF through numerous compounds,targets and pathways.The inhibition of osteoclast differentiation may play an important role in the mechanism of PT against OF.