补骨脂二氢黄酮甲醚对破骨细胞分化的影响及机制

Study on the effect of bavachinin on osteoclast differentiation and its mechanism

目的 探究补骨脂中二氢黄酮类化合物在破骨细胞分化中的作用。方法 采用RANKL+M-CSF诱导骨髓单核细胞向破骨细胞分化,在分化过程中给予补骨脂二氢黄酮、异补骨脂二氢黄酮、补骨脂二氢黄酮甲醚,干预3 d后,根据各组细胞的抗酒石酸酸性磷酸酶活性计算EC_(50),筛选抑制作用最强的化合物进行下一步试验。采用Western blot分析补骨脂二氢黄酮甲醚干预下的破骨细胞中NF-κB信号通路、PI3k/AKT信号通路、MAPK信号通路的关键蛋白的水平,探索其可能的作用机制。结果 补骨脂二氢黄酮、异补骨脂二氢黄酮、补骨脂二氢黄酮甲醚的EC_(50)分别为15.89、15.18、12.39μmol/L。补骨脂二氢黄酮甲醚可以明显下调破骨细胞分化过程中p65、AKT、p38、JNK、ERK的磷酸化水平。结论 补骨脂二氢黄酮、异补骨脂二氢黄酮、补骨脂二氢黄酮甲醚中补骨脂二氢黄酮甲醚对破骨细胞的分化抑制作用最强,其作用机制与抑制NF-κB信号通路、PI3k/AKT信号通路、MAPK信号通路的激活有关,为补骨脂二氢黄酮甲醚临床应用于骨质疏松症的治疗提供参考。

Objective To explore the role of dihydroflavonoids of psoraleae fructus in the differentiation of osteoclasts.MethodsRANKL+M-CSF was used to induce bone marrow monocytes to differentiate into osteoclasts.During the differentiation process,isobavachin,bavachin,bavachinin were additioned for intervention.After 3 days,the EC_(50) was calculated according to the anti-tartrate acid phosphatase activity of each group of cells.The compound with the strongest inhibitory effect was selected for the next test.Western blotting was used to analyze the levels of key proteins in the NF-κB signaling pathway,PI3k/AKT signaling pathway,and MAPK signaling pathway in osteoclasts under the intervention of bavachinin.Results The EC_(50) of isobavachin,bavachin,and bavachinin was 15.89,15.18,and 12.39μmol/L,respectively.Bavachinin significantly down-regulated the phosphorylation levels of p65,AKT,p38,JNK,and ERK during osteoclast differentiation.Conclusion Bavachinin has the strongest inhibitory effect on osteoclast differentiation.Its mechanism of action is related to the inhibition of the activation of NF-κB signaling pathway,PI3k/AKT signaling pathway,and MAPK signaling pathway.This study provides reference for the clinical application of bavachinin in the treatment of osteoporosis.