松果菊苷通过激活ERK/BMP-2信号通路促进大鼠的成骨细胞增殖观察

摘要

目的：考察松果菊苷对大鼠成骨细胞增殖的影响及作用机制。方法将不同浓度的松果菊苷（0.01，0.1，1.0，10.0，100 nmol/L）及MAPK/ERK信号通路抑制剂PD98059（20μmol/L）与成骨细胞共培养12，24，36，48，60 h后，采用MTT法检测细胞增殖情况。酶联免疫吸附法（ELISA）和qRT-PCR法检测细胞上清中和成骨细胞中骨钙素（BGP）和骨形态发生蛋白（BMP-2）的表达水平。Western blotting检测成骨细胞中Smad4、ERK1/2和p-ERK1/2的蛋白表达水平。结果松果菊苷能够促进大鼠成骨细胞的增殖，而且能诱导ERK 的磷酸化从而激活 MAPK/ERK 信号通路，并提高成骨细胞中 BMP-2和 Smad4的表达而激活BMP/Smad信号通路。加入PD98059后，松果菊苷诱导的BMP-2和Smad4表达及成骨细胞增殖均受到抑制。结论松果菊苷可通过激活ERK/BMP-2信号通路促进大鼠的成骨细胞增殖。%ObjectiveTo investigate the effect of echinacoside(ECH)on cell proliferation of rat osteoblast and its mechanism.MethodThe osteoblast was incubated with different concentration of ECH(0.01、0.1、1.0、10.0、100 nmol/L)or PD98059(20 μmol/L)which is an inhibitor of MAPK/ERK for 12、24、36、48 or 60 h. MTT method was used for the detection of cell proliferation. The protein and mRNA level of BGP and BMP-2 in supernatant and cells was measured by ELISA and qRT-PCR. Expression of smad4、ERK1/2 and p-ERK1/2 was detected by western blotting. ResultsThe cell proliferation of osteoblast was promoted by ECH. ECH induced expression of BMP-2 and smad4 to activate BMP/smad pathway,and also promoted the phosphorylation of ERK1/2 to activate MAPK/ERK pathway. While the activation of BMP/smad pathway was inhibited by PD98059.ConclusionsEchinacoside promotes cell proliferation of rat osteoblast through activating of ERK/BMP-2 signaling pathway.