从彝药斯赤列中寻找具有NOX4抑制活性的抗骨质疏松活性化合物

衰老造成的NOX4表达上升诱导ROS大量生成是导致成骨与破骨分化失衡的重要原因。斯赤列是彝族传统用药，广泛用于治疗骨质疏松、骨折等疾病。我们前期研究发现斯赤列提取物中含有大量三萜类化合物，且具有较好的促进成骨细胞分化、改善骨质疏松的作用并显示出一定的NOX4抑制活性，但具体作用物质基础与作用机制不明。因此，本项目拟建立高表达NOX4的HEK293细胞模型并结合色谱方法，系统分离彝药斯赤列抑制NOX4活性的化合物，以氧化应激致大鼠BMSCs衰老模型成骨细胞和破骨前体细胞模型进一步筛选出具有调节成骨和破骨功能的活性化合物，再以自发性老年骨质疏松SAMP6小鼠模型考察其体内活性，筛选出活性优良的化合物，并进行构效关系分析，采用HPLC、LC-MS法建立药材的指纹图谱。通过本项目研究，有望从彝药斯赤列中筛选出以NOX4为靶点的抗骨质疏松活性化合物，为我省丰富的彝药资源提供系统的基础研究示范。

Increasement of NOX4 expression induced the boost of ROS, which was the important factor to destroy the balance of osteoblast and osteoclast differentiation. Sichilie was used to treat traumatic injury, bone fractures, joint pain, et al. As an ethnic and folk herbal medicines, the raw material resources of the plant is abundant. We investigated the chemical constituents and the activities for promoting bone fracture healing of the extracts of Sichilie in previous research. Some triterpenoids and other chemical constituents were isolated and identified. Although the effective phytochemical constituents were not discovered, it’s very interesting that the extracts of Sichilie was active to preventing osteoporosis by inhibiting the upregulation of NOX4. Base on the previous studies and abundent valuable references, the research methods of phytochemical compositions combining activities were projecting to know the active principles for preventing osteoporosis by inhibiting the upregulation of NOX4. Chromatograph and spectrascopy methods would be utilized to isolated and identified the active chemical constituents. By the means of the overexpression of NOX4 in HEK293 cell, the activies of compounds inhibiting NOX4 would be investigated. Furthermore, osteoclast precursors cells and osteoblast cells from the aging rats model with BMSCs would play a role finding the active compounds which could stimulate osteoblast differentiation and inhibit osteoclast differentiation. Then repeat-dose oral studies of these effective compounds were done in senescene accelarated mice (SAMP6) to find the valuable compounds and demonstrate the phytochemical structure-efficacy relationship. To improve the quality standard of the herbal material, HPLC and LC-MS would be used to analyse the fingerprint chromatograms. Above all, the project was significant to find the compounds inhibiting NOX4 from the extracts of Sichilie, which could set an example for the systematic study of the Yi People’s herbal mdedicine.