红曲有效成分对体外培养成骨细胞增殖分化及矿化功能的影响

摘要

[Objective] To ascertain effective ingredients of Hong Qu on proliferation,differentiation and mineralization functions of osteoblasts by cytological tests, on purpose of providing a cytological foundation for treating osteoporosis by Chinese medicine. [Methods] 40 SD rats were involved,and were randomly divided into 4 groups,as the blank group, the premarin group, the α-D3 group,and the Hong Qu effective ingredients group. All the rats were lavaged by lOml/kg respectively,with saline,premarin solution, orD3 solution and Hong Qu extract solution. After a 10-day lavage,killed the rats and took the rats'blood to prepare medicated serum. Osteoblasts were obtained from calvaria of 24h-newborn rats, and were cultured in serum-containing mediums above-mentioned. MMT assay, P-nitrophenyl phosphate method (PNPP), BMP-2 immunohistochemical staining and Alizarin red staining (ARS)were used for detecting effects of Hong Qu extract on proliferation, ALP expression,BMP-2 expression and mineralized nodule formation of osteoblasts in vitro,on purpose of ascertaining effective ingredients of Hong Qu in treating osteoporosis. [Results] Medicated serum of Hong Qu effective ingredients could obviously increase the number of osteoblasts, the expression of ALP and the formation of mineralized nodules. Compared to group A,group D had higher positive rate of BMP-2. [Conclusion] The speculation of the effect of effective ingredients of Hong Qu in treating osteoporosis was based on its function of promoting proliferation, differentiation, mineralization and BMP-2 expression of osteoblasts.%[目的]通过红曲有效成分对体外培养成骨细胞增殖、分化及矿化功能影响的研究，对有效成分抗骨质疏松症的作用机理进行初步探讨。[方法]取成年SD大鼠40只，随机分为空白组、倍美力组、α-D3组、红曲有效成分组4组，每组10只，分别予以生理盐水、倍美力水溶液、α-D3水溶液及红曲有效成分水溶液灌胃，10d后，制备含药血清；将从出生24h内的新生大鼠颅骨取材的成骨细胞培养于上述含药血清培养基中，应用MTT法、对硝基苯磷酸盐法、BMP-2免疫组化染色及茜素红染色方法观察红曲有效成分对体外培养成骨细胞的增殖、碱性磷酸酶表达、BMP-2表达及矿化结节形成的影响。[结果]红曲有效成分组的成骨细胞数量、碱性磷酸酶的表达水平及矿化结节的形成明显增加，而且发现BMP-2阳性率高于空白组。[结论]红曲有效成分能治疗骨质疏松症，其机制可能是通过促进成骨细胞增殖、分化、矿化及BMP-2表达来实现的。