黄瓜籽总皂苷对MC3T3-E1细胞分化及SPARC、OPG/RANKL表达的影响

目的观察黄瓜籽总皂苷提取物(cucumber seed saponins,CSS)对小鼠成骨细胞MC3T3-E1增殖、分化和矿化的影响,以及与骨质疏松相关的SPARC、OPG/RANKL/RANK信号通路的作用。方法通过MTT实验、碱性磷酸酶(alkaline phosphatase,ALP)活性检测、茜素红染色,考察不同浓度CSS对MC3T3-E1细胞增殖、分化及矿化的影响;采用RT-PCR方法检测SPARC、OPG/RANKL mRNA表达水平; Western blot检测SPARC、OPG/RANKL的蛋白表达量。结果与阳性对照组相比,CSS能明显促进MC3T3-E1细胞增殖(P0.05),CSS高、中剂量组能明显提高MC3T3-E1细胞ALP活性及钙化结节数量(P0.05);与空白组相比,CSS不同剂量组均明显上调SPARC、OPG/RANKL mRNA及蛋白表达水平(P0.05)。结论黄瓜籽总皂苷能够促进成骨细胞MC3T3-E1的增殖、分化及矿化能力,并通过上调SPARC、OPG/RANKL的表达水平提高MC3T3-E1细胞的成骨能力。

The effect of cucumber seed saponins on the differentiation of osteoblast and the expression of SPARC and OPG/RANKL in MC3T3-E1 cells

Objective To investigate the effect of cucumber seed saponins (CSS) on the proliferation, differentiation, and mineralization in osteoblastic cell MC3T3-E1, as well as on the regulation of the SPARC, OPG/RANKL/RANK signal pathways. Methods The changes of cell proliferation, differentiation, and mineralization were determined using MTT test, alkaline phosphatase (ALP) activity, and the Alizarin red staining calcified nodules. The expressions of SPARC and OPG/RANKL mRNA in the cells were detected with RT-PCR. The protein expressions of SPARC and OPG/RANKL were observed with Western blotting. Results In MC3T3-E1 cells, compared with the positive control group, medium containing CSS significantly increased in cell viability (P<0.05). High and medium doses of CSS significantly promoted ALP activity and amount of calcified nodules (P<0.05). The different concentrations of CSS up-regulated SPARC and OPG/RANKL mRNA and protein expression, and the promoting level of was significantly higher than those in the control group (P<0.05). Conclusion CSS promotes the proliferation, differentiation, and mineralization of MC3T3-E1 cells. CSS increases the osteogenic ability of MC3T3-E1 cells by up-regulating the expression levels of SPARC and OPG/RANKL.