蛇床子对肾阳虚骨质疏松大鼠的治疗作用与机制研究

Effect and Mechanism of Cnidium monnieri on Osteoporosis in Kidney Yang Deficiency-induced Osteoporotic Rats

[目的]探索蛇床子对肾阳虚骨质疏松大鼠的骨保护作用与分子机制。[方法]SD大鼠分为正常、模型、蛇床子提取物与蛇床子素组。除正常组外,其余大鼠每日皮下给予氢化可的松,给药组每日灌胃给药,连续3周。大鼠麻醉后采集股骨,其中左侧股骨进行抗酒石酸磷酸酶染色、骨钙素与转化生长因子-β(transforming growth factor-β,TGF-β)免疫组化染色以及骨密度检测,右侧股骨进行实时荧光定量聚合酶链式反应与免疫印迹检测靶基因和蛋白表达。[结果]氢化可的松可显著促进大鼠破骨细胞分化与成熟,抑制成骨细胞生成,降低骨小梁生成,降低骨密度,最终诱导大鼠肾阳虚骨质疏松发生。生物信息预测结果提示,TGF-β信号通路是蛇床子潜在候选靶标通路。蛇床子可显著减轻由氢化可的松诱导的破骨细胞分化与成熟现象,并促进成骨细胞分化与成熟,从而减轻氢化可的松诱发的大鼠骨质疏松。蛇床子提取物可显著增强股骨中因氢化可的松诱导的TGF-β基因下调。[结论]蛇床子可通过调控TGF-β信号通路,抑制肾阳虚大鼠骨质疏松。

[Objective]To investigate the osteoprotective effect and molecular mechanism of Cnidium monnieri on kidney Yang deficiency-induced osteoporosis in rats.[Methods]SD rats were divided into normal,model,Cnidium monnieri extract and osthole groups.The rats except for normal group were given hydrocortisone subcutaneously every day.The rats were given Cnidium Monnieri extract,osthole or vehicle intragastric daily for 3 weeks.After anesthesia,rat femurs were collected.Tartrate phosphatase staining,transforming growth faction-β(TGF-β)immunohistochemical staining and bone mineral density detection were performed on the left femur,while Real-time quantitative polymerase chain reaction and western blot detection of target gene and protein expression were performed on the right femur.[Results]Hydrocortisone has been shown to significantly enhance the differentiation and maturation of rat osteoclasts,while inhibiting the formation of osteoblasts.This results in a reduction in the formation of bone trabeculae and ultimately induces osteoporosis in rats with kidney-yang deficiency.The results of bioinformatics analysis suggested that TGF-βsignaling pathway was the potential candidate target pathway for Cnidium monnieri.Cnidium monnieri extract and osthole significantly reduced the differentiation and maturation of osteoclasts,promoted the differentiation and maturation of osteoblasts,and ultimately inhibited the osteoporosis in rats induced by hydrocortisone.The extract of Cnidium monnieri and osthole significantly increased the downregulation of TGF-βgene induced by hydrocortisone in femur.[Conclusion]Cnidium monnieri and osthole can inhibit osteoporosis in kidneyYang deficiency rats through TGF-βsignaling pathway.