小分子肽对成骨前体细胞MC3T3-E1骨保护素和核转录因子κB受体活化因子配体表达的影响 |
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作者姓名: | 吴昆鹏 杨巧珠 马凤燕 戴永武 刘 义 |
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作者单位: | 1广东医学院第一临床学院,广东省湛江市 524023
2 广东天然药物研究与开发重点实验室,广东省湛江市 524023 |
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基金项目: | 广东省科技计划项目(2010B090400463,2009B030801339,2010B090400463);广东省教育厅大学生创新实验项目(KY1007);广东省医学院大学生课外科研(KY0701)。 |
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摘 要: | 背景:体内实验显示,小分子肽能明显增加去卵巢大鼠的骨钙含量,使其骨密度增加,能很好地预防骨质疏松。同时体外实验显示,小分子肽能促进小鼠成骨细胞和成骨前体细胞MC3T3-E1增殖、分化、矿化,并且可能是通过抑制核转录因子 p50和p65的表达来起作用。而小分子肽对骨保护素/核转录因子κB受体活化因子配体的影响尚不明确。
目的:观察小分子肽对MC3T3-E1在增殖、分化、矿化过程中骨保护素和RANKL表达的影响。
方法:以体积分数10%胎牛血清的DMEM培养液为空白对照组,50,100 mg/L质量浓度小分子肽作用小鼠成骨前体细胞MC3T3-E1,分别于作用3,6,12,18,24,30 d后,收集细胞提取蛋白,Western Blot检测骨保护素和核转录因子κB受体活化因子配体蛋白的表达。
结果与结论:50,100 mg/L小分子肽作用MC3T3-E1后能明显促进作用骨保护素的表达(P < 0.01),而对核转录因子κB受体活化因子配体无明显影响。小分子肽作用后MC3T3-E1中骨保护素/核转录因子κB受体活化因子配体的比值要明显高于空白对照组(P < 0.01)。因此,认为小分子肽可以通过增加骨保护素的表达来影响骨保护素/核转录因子κB受体活化因子配体系统,间接地抑制破骨细胞的数量和功能。
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关 键 词: | 小分子肽 骨保护素 核转录因子&kappa B受体活化因子配体 成骨前体细胞 组织构建 |
收稿时间: | 2011-01-21 |
Effect of the small molecular weight peptide on expression of osteoprotegerin and receptor activator of nuclear factor-kappa B ligand in MC3T3-E1 cells |
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Authors: | Wu Kun-peng Yang Qiao-zhu Ma Feng-yan Dai Yong-wu Liu Yi |
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Institution: | 1The First Clinical Institute, Guangdong Medical College, Zhanjiang 524023, Guangdong Province, China
2Guangdong Key Laboratory for Research and Development of Nature Drugs, Zhanjiang 524023, Guangdong Province, China |
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Abstract: | BACKGROUND:The in vivo experiment indicated that the small molecular weight peptide (KP) can treat osteoporosis in ovariectomized rat by increasing the level of calcium (Ca) and bone mineral density. The in vitro experiment indicated that KP can induce the proliferation, differentiation and mineralization of MC3T3-E1 cells by inhibiting nuclear factor-κ B p50 and p65. It is unknown whether KP affects the expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) in osteoblasts.
OBJECTIVE:To investigate the effect of KP on the expression of OPG and RANKL in MC3T3-E1.
METHODS:MC3T3-E1 were cultured with various concentrations of KP (0, 50 and 100 mg/L). Osteoblasts cultured without KP were assigned as control. Collected cells and analyzed the expression of OPG and RANKL with Western Blot after treating for 3, 6, 12, 18, 24, and 30 days.
RESULTS AND CONCLUSION:After 50 mg/L and 100 mg/L KP treatment, the OPG expressions in MC3T3-E1 cells were gradually increased (P < 0.01), and KP had no effect on the expression of RANKL. Compared with control, KP obviously increased the OPG/RANKL ratio (P < 0.01). Therefore, we think that KP affects the system of OPG/RANKL and indirectly inhibits osteoclast number and functions by increasing the expression of OPG, but RANKL. |
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