环淫羊藿素逆转糖皮质激素所致骨质疏松的机理研究

汪华君; 乔晨; 赵蓉; 尹江宁

环淫羊藿素逆转糖皮质激素所致骨质疏松的机理研究

江苏大学附属医院，江苏镇江 212001

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出版历程
- 刊出日期: 2019-11-10

Study on the Mechanism of Circular-Icaritin Against Glucocorticoid-Induced Osteoporosis

摘要

摘要: 目的阐明环淫羊藿素(CICT)逆转糖皮质激素所致骨质疏松作用，并探讨其作用机制。方法采用泼尼松龙诱发斑马鱼骨质疏松模型，茜素红染色后，荧光倒置显微镜观察CICT对斑马鱼头骨染色情况、计算染色面积和累计光密度；ICP-MS测定各组斑马鱼中的钙和磷元素含量；采用分子对接技术模拟并预测CICT与BMPs的相互作用；采用qPCR检测Runx 2和AKP基因的表达量。结果与空白组比，经泼尼松龙(25 μmol/L)培养后，可显著降低斑马鱼骨累积光密度和头骨矿化面积(P<0.01)，且斑马鱼的Ca和P水平显著降低(P<0.01)，Runx 2和AKP基因表达量显著降低(P<0.01)；与泼尼松龙组相比，经不同剂量的CICT(0.1、1.0、10.0 μmol/L)干预后，斑马鱼的头骨累积光密度值和骨矿化面积均有显著的提高(P<0.01)，斑马鱼Ca和P水平均显著提高(P<0.01)；采用分子对接技术研究表明，CICT可与靶蛋白BMPs(BMP-2/BMP-4)稳定对接，对接得分分别为-5.493 256 09和-5.993 616 58；Runx 2和AKP基因表达量有显著增加(P<0.01)。结论 CICT能逆转糖皮质激素所致骨质疏松，这一作用可通过CICT与BMPs蛋白靶点结合，并调节BMPs信号通路而促进成骨分化而发挥抗骨质疏松作用。
- 环淫羊藿素 /
- BMP-2 /
- BMP-4 /
- 泼尼松龙 /
- 斑马鱼 /
- 骨质疏松 /
- 分子对接
Abstract: OBJECTIVE To elucidate the reversal effect of cyclic-icaritin(CICT) on prednisolone -induced osteoporosis， and to explore its mechanism of action with BMPs signaling pathway. METHODS Prednisolone-induced osteoporosis model of zebrafish was used. The stained area， cumulative optical density， calcium and phosphorus contents of the skull of zebrafish were observed by fluorescence inverted microscope (×100). qPCR was used to quantitatively detect the expression of Runx 2 and AKP genes. RESULTS Compared with the control group， the cumulative optical density and mineralized area of zebrafish skull significantly decreased (P<0.01)，the Ca and P levels significantly decreased (P<0.01)， the expression of Runx 2 and AKP genes decreased significantly (P<0.01) after incubation with prednisolone (25 μmol/L).After the intervention with different doses of CICT (0.1， 1.0， 10.0 μmol/L)，the cumulative optical density and bone mineralization area of zebrafish skull significantly increased (P<0.01)， and the levels of Ca and P significantly increased (P<0.01). Molecular docking techniques showed that CICT could stably dock with target protein BMPs (BMP-2/BMP-4)， and the docking scores were - 5.49325609 and - 5.99361658， respectively. The expressions of Runx-2 and AKP genes significantly increased (P<0.01). CONCLUSION CICT can reverse glucocorticoid-induced osteoporosis， which plays an anti-osteoporosis role by binding CICT to BMPs protein target and regulating BMPs signaling pathway to promote osteogenic differentiation.
- cyclic-icaritin /
- BMP-2 /
- BMP-4 /
- prednisolone /
- zebrafish /
- osteoporosis /
- molecular docking

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