Network Pharmacology and Experimental Validation of Sanqi-Danshen in the Treatment of Osteoporosis

Osteoporosis (OP) is a systemic skeletal disorder with a systemic metabolic bone disease including reduced bone mineral density (BMD), deteriorated skeletal micro-architecture and ultimately lead to fragility fracture. Sanqi is the dry roots of Panax notoginseng (Burk.) F. H. Chen, and Danshen is the dry root and rhizome of Salvia miltiorrhiza Bge. The two are considered to be a pair of drugs in OP. However, the mechanism has not been fully clarified. Herein, a systematic study based on network pharmacology was designed to elucidate this pair of drugs. The pharmacological system generated 66 bio-active ingredients and 213 targets from Sanqi-Danshen. Additionally, 1762 targets have entered the analysis, and 96 candidates of Sanqi-Danshen on OP were selected. Quercetin and luteolin might be the essential components of Sanqi-Danshen for OP therapy in network analysis. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) indicated that Sanqi-Danshen can have effects on bone metabolism, antioxidant, anti-inflammatory activity for OP treatment through AGE-RAGE signaling pathway in diabetic complications, Leishmaniasis, IL- 17 signaling pathway ( p < 0.05, q < 0.05). PPI network and sub-networks identified 5 hub genes including NTRK1, TP53, EGFR, CUL3 and HSP90AA1 were exerting potential therapeutic effects in OP. Different concentrations of quercetin and luteolin could lead to raw264.7 cells cytotoxicity and apoptosis. Moreover, the BLI and RT-PCR also confirmed that quercetin and luteolin have a good affinity to IL-6 and these compounds would regulate the mRNA expression of NTRK1, Caspase-3, IL-6, ESR1 and TNF- α . In summary, the drug pairs of Sanqi and Danshen show a multi-compound, multi-target to OP treatment, and our research would provide a scientific basis and guidance for further clinical applications and mechanism action.