Department of Orthopedics, Affifiliated Zhongshan Hospital of Dalian University, Dalian, 116001, China; Ansteel Group Hospital, Anshan, 114002, China.
Ansteel Group Hospital, Anshan, 114002, China.
J Ethnopharmacol. 2022 Sep 15;295:115354. doi: 10.1016/j.jep.2022.115354. Epub 2022 May 14.
Weishi Huogu I (WH I) capsules, developed through traditional Chinese medicine, have been used to treat clinical osteonecrosis of the femoral head (ONFH) for decades. However, the mechanisms have not been systematically studied.
In this study, the mechanisms of WH I capsules used in treating ONFH were examined through a systems pharmacology strategy, and one mechanism was validated with in vitro experiments.
WH I capsules compounds were identified by screening databases; then, a database of the potential active compounds was constructed after absorption, distribution, metabolism and excretion (ADME) evaluation. The compounds were identified through a systematic approach in which the probability of an interaction of every candidate compound with each corresponding target in the DrugBank database was calculated. Gene Ontology (GO) and pathway enrichment analyses of the targets was performed with the Metascape and KEGG DISEASE databases. Then, a compound-target network (C-T) and target-pathway network (T-P) of WH I capsule components were constructed, and network characteristics and related information were used for systematically identifying WH I capsule multicomponent-target interactions. Furthermore, the effects of WH I capsule compounds identified through the systematic pharmacology analysis of the osteogenic transformation of human umbilical mesenchymal stem cells (HUMSCs) were validated in vitro.
In total, 152 potentially important compounds and 176 associated targets were identified. Twenty-two crucial GO biological process (BP) or pathways were related to ONFH, mainly in regulatory modules regulating blood circulation, modulating growth, and affecting pathological processes closely related to ONFH. Furthermore, the GO enrichment analysis showed that corydine, isorhamnetin, and bicuculline were enriched in "RUNX2 regulates osteoblast differentiation", significantly increased alkaline phosphatase activity and calcium deposition and upregulated runt-related transcription factor 2 mRNA and protein expression and osteocalcin mRNA expression in HUMSCs, suggesting that these compounds promoted the mesenchymal stem cell (MSC) osteogenic transformation.
The study showed that the pharmacological mechanisms of WH I capsule attenuation of ONFH mainly involve three therapeutic modules: blood circulation, modulating growth, and regulating pathological processes. The crosstalk between GOBPs/pathways may constitute the basis of the synergistic effects of the compounds in WH I capsules in attenuating ONFH. One of the pharmacological mechanisms in the WH I capsule effect on ONFH involves enhancement of the osteogenic transformation of MSCs, as validated in experiments performed in vitro; however, more mechanisms should be validated in further studies.
卫食活骨 I（WH I）胶囊是一种中药，已被用于治疗临床股骨头坏死（ONFH）数十年。然而，其机制尚未得到系统研究。
本研究采用系统药理学策略研究 WH I 胶囊治疗 ONFH 的作用机制，并通过体外实验验证其中一种机制。
通过筛选数据库鉴定 WH I 胶囊化合物；然后，经过吸收、分布、代谢和排泄（ADME）评估，构建潜在活性化合物数据库。通过系统方法鉴定化合物，计算候选化合物与 DrugBank 数据库中每个对应靶标的相互作用概率。使用 Metascape 和 KEGG DISEASE 数据库对靶点进行基因本体论（GO）和途径富集分析。然后，构建 WH I 胶囊成分的化合物-靶点网络（C-T）和靶点-通路网络（T-P），并利用网络特征和相关信息系统地识别 WH I 胶囊多成分-靶点相互作用。此外，通过对人脐带间充质干细胞（HUMSCs）成骨转化的系统药理学分析，验证 WH I 胶囊化合物的体外作用。
共鉴定出 152 种潜在重要化合物和 176 个相关靶点。与 ONFH 相关的 22 个关键 GO 生物过程（BP）或途径主要集中在调节血液循环、调节生长、影响与 ONFH 密切相关的病理过程的调节模块中。此外，GO 富集分析表明，延胡索乙素、异鼠李素和荷包牡丹碱富集于“RUNX2 调节成骨细胞分化”，显著提高碱性磷酸酶活性和钙沉积，并上调 HUMSCs 中 runt 相关转录因子 2 mRNA 和蛋白表达以及骨钙素 mRNA 表达，提示这些化合物促进间充质干细胞（MSC）成骨转化。
本研究表明，WH I 胶囊减轻 ONFH 的药理机制主要涉及三个治疗模块：血液循环、调节生长和调节病理过程。GO/BP 途径之间的串扰可能构成 WH I 胶囊减轻 ONFH 协同作用的基础。WH I 胶囊对 ONFH 作用的药理机制之一涉及增强 MSC 的成骨转化，这在体外实验中得到了验证；然而，还需要更多的机制在进一步的研究中得到验证。
