Integrated network pharmacology and pharmacological evaluation to elucidate the effect and mechanism of the novel combination PMS in treatment of osteoporosis

Psoralen, magnoflorine and sweroside were main active components of Xian-Ling-Gu-Bao capsule. PMS (a combination of psoralen, magnoflorine and sweroside at extremely low concentration), exhibited excellent promoting osteoblastic activity on pre-osteoblastic MC3T3-E1 cells in our previous work, and has been filed as a patent in China. However, the effect and mechanism of PMS for anti-osteoporosis at present remain unclear. Therefore, further exploring the potential anti-osteoporosis mechanism of PMS to strongly provide more evidences for developing as a novel derivative drug. In the present study, in vivo experiments were conducted to evaluate anti-osteoporosis action of PMS by using zebrafish model, and PMS treatment could significantly inhibit dexamethasone-induced osteoporosis in zebrafish. Subsequently, the data of network pharmacology and the intracellular concentrations of each ingredient further analyzed GPR30/PI3K/AKT signaling pathway probably involved in PMS-induced osteoblastic activity. Importantly, the results of western blot exhibited that PMS could increase the expressions of the membrane receptor GPR30, p-PI3K/PI3K and p-AKT/AKT on MC3T3-E1 cells, while PMS-induced osteoblastic differentiation and these protein expressions on MC3T3-E1 cells were partially blocked by the GPR30 antagonist. Collectively, our study for the first time proposed that the novel combination PMS might increase osteoblastic activity to improve osteoporosis via activating GPR30/PI3K/AKT signaling pathway, which offered a unique perspective on the mechanistic pathways of PMS in alleviating osteoporosis.