[Advantageous therapeutic pathways and mechanisms of Jianpi Huogu Formula in treating steroid-induced osteonecrosis of femoral head based on multi-source heterogeneous data integration of disease-syndrome-formula framework].

Based on integrated analysis of multi-source heterogeneous biomedical data combined with animal experimental validation, this study systematically explored the advantageous therapeutic pathways and molecular mechanisms of Jianpi Huogu Formula(JPHGF) in treating steroid-induced osteonecrosis of the femoral head(SONFH). First, the candidate active components and targets of JPHGF were obtained from the Encyclopedia of Traditional Chinese Medicine(ETCM v 2.0). Meanwhile, the Human Phenotype Ontology(HPO) database was used to identify potential genes associated with the corresponding syndrome pattern. Finally, clinical transcriptomic data were analyzed to obtain relevant targets for the phlegm-blood stasis blocking collateral syndrome of SONFH. The intersection of these three types of targets was used to construct a multidimensional "drug-ingredient-disease-syndrome" network. The STRING database was employed for protein-protein interaction(PPI) network analysis, and the Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis of core targets was performed via the DAVID platform to predict key biological processes and signaling pathways. Pharmacodynamic and mechanistic validation was subsequently conducted using a rat model of SONFH with phlegm-blood stasis obstructing collateral syndrome. Data integration and mining yielded a "disease and syndrome gene-formula and drug target" network containing 146 core targets. Pathway enrichment analysis indicated that JPHGF primarily regulates lipid-coupled intervention in the SONFH phlegm-blood stasis blocking collateral phenotype through the purinergic receptor P2Y12(P2RY12)-phosphoinositide 3-kinase(PI3K)-serine/threonine kinase(Akt)-fibrinogen gamma chain(FGG) signaling pathway. Hematoxylin-eosin(HE) staining revealed that rats with SONFH of the phlegm-blood stasis blocking collateral type exhibited reduced calcification in femoral head tissues, loosely arranged trabeculae, hypoplastic osteocytes, significantly increased lacunae, and elevated numbers of intramedullary adipocytes. Lipid profile analysis showed elevated triglyceride, total cholesterol, and low-density lipoprotein(LDL) levels, along with decreased high-density lipoprotein(HDL). Coagulation assays demonstrated shortened activated partial thromboplastin time(APTT) and prothrombin time(PT), as well as prolonged thrombin time(TT) and increased plasma fibrinogen levels. Immunohistochemical staining showed that compared with the normal group, the model group exhibited significantly increased positive expression areas of P2RY12, phosphorylated PI3K(p-PI3K), phosphorylated Akt(p-Akt), and FGG proteins in femoral head tissues(P