Table2_Bushen huoxue decoction inhibits RANKL-stimulated osteoclastogenesis and glucocorticoid-induced bone loss by modulating the NF-κB, ERK, and JNK signaling pathways.DOCX

Glucocorticoid-induced osteoporosis (GIOP) is the most common form of secondary osteoporosis, which is caused by a disorder in bone metabolism due to excessive activation of osteoclasts. Bushen Huoxue decoction (BHD) is an herbal formula with multiple pharmacological effects, including anti-inflammatory, antioxidant activity and stem cell migration promotion. However, the effect of BHD on osteoclastogenesis has not been reported. In this study, we aimed to elucidate the effect of BHD on RANKL-stimulated osteoclastogenesis and explored its underlying mechanisms of action in vitro. Our results show that BHD had no effect on BMMs and RAW264.7 cells viability, but inhibited RANKL-induced osteoclast formation in vitro. Furthermore, BHD attenuated RANKL-induced NF-κB, ERK, and JNK signaling. The attenuation of NF-κB, ERK, and JNK activation were enough to impede downstream expression of c-fos and NFATc1 and related specific genes. Meanwhile, we investigated the therapeutic effect of BHD on glucocorticoid-induced osteoporosis (GIOP) mice. The result indicated that BHD prevents glucocorticoid-induced osteoporosis and preserves bone volume by repressing osteoclast activity. Collectively, BHD shows significant osteoclast inhibition and holds great promise in the treatment of osteoporosis.

糖皮质激素诱导性骨质疏松（glucocorticoid-induced osteoporosis, GIOP）是最常见的继发性骨质疏松类型，其发病机制为破骨细胞过度活化引发骨代谢紊乱。补肾活血汤（Bushen Huoxue decoction, BHD）是一种具有多重药理作用的中药方剂，涵盖抗炎、抗氧化活性及促进干细胞迁移等功效。然而，补肾活血汤对破骨细胞生成的调控作用尚未见报道。本研究旨在阐明补肾活血汤对核因子κB受体活化因子配体（RANKL）刺激的破骨细胞生成的影响，并体外探究其潜在作用机制。研究结果显示，补肾活血汤对骨髓源性巨噬细胞（bone marrow-derived macrophages, BMMs）与RAW264.7细胞的存活率无显著影响，但可在体外抑制RANKL诱导的破骨细胞形成。此外，补肾活血汤可减弱RANKL激活的核因子κB、细胞外调节蛋白激酶（ERK）及c-Jun氨基末端激酶（JNK）信号通路。对核因子κB、ERK及JNK活化的抑制作用，足以阻碍下游c-fos、活化T细胞核因子1（NFATc1）及其相关特异性基因的表达。同时，本研究还探究了补肾活血汤对GIOP模型小鼠的治疗效果，结果表明，补肾活血汤可通过抑制破骨细胞活性，改善糖皮质激素诱导性骨质疏松并维持骨量。综上，补肾活血汤具有显著的破骨细胞生成抑制活性，在骨质疏松症的临床治疗中具备良好的应用前景。