林蛙油提取物对过氧化氢诱导MC3T3-E1损伤的保护

Protection of rana oil extract on hydrogen peroxide-induced MC3T3-E1 injury

中文关键词:  林蛙油提取物  成骨细胞MC3T3-E1  骨质疏松  过氧化氢

英文关键词:rana oil extract  osteoblast MC3T3-E1  osteoporosis  hydrogen peroxide

中文摘要:

      目的 研究林蛙油提取物(rana oil extract, ROE)对过氧化氢致小鼠成骨细胞(MC3T3-E1)损伤的保护作用。方法 采用MTT方法检测细胞活力, 筛选林蛙油提取物促MC3T3-E1细胞生长的最佳给药浓度和时间, Western blotting法检测骨形态发生蛋白-2(BMP2)、碱性磷酸酶(alkaline phosphatase, ALP)、骨钙素(osteocalcin)、Runt相关转录因子2(Runx2)蛋白表达。应用逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction, RT-PCR)检测骨生长相关基因的表达。结果 经过MTT法筛选, 400 μg/mL、24 h为ROE的最佳给药浓度及时间, 与空白对照组比较, 模型对照组MC3T3-E1细胞ALP蛋白表达水平降低(P<0.05), Runx2、BMP2、Osteocalcin表达明显降低(P<0.01)。与模型对照组比较, ROE给药组中ALP、BMP2蛋白表达水平升高(P<0.05) Osteocalcin、Runx2蛋白表达水平明显升高(P<0.01), 与空白组比较, 模型组ALP、Runx2、BMP2基因表达均下调(P<0.05), Osteocalcin 显著下调(P<0.01); 与模型组比较, ROE给药组ALP、Runx2基因表达均上调(P<0.05), BMP2、Osteocalcin基因表达显著下调(P<0.01)。结论 ROE可以调控骨代谢基因和蛋白的表达水平, 对H2O2损伤的MC3T3-E1老年型骨质疏松模型有一定的保护作用。

英文摘要:

      Objective To study the protective effect of rana oil extract (ROE) on the injury of mouse osteoblasts (MC3T3-E1) induced by hydrogen peroxide. Methods MTT assay was used to detect cell viability. The best concentration and time of MC3T3-E1 cell growth were screened by rana oil extract. Bone morphogenetic protein-2 (BMP2) and alkaline phosphatase (ALP) were detected by Western blotting. Osteocalcin and Runt-related transcription factor 2 (Runx2) protein expression. The expression of bone growth related genes was detected by RT-PCR. Results After MTT assay, 400 μg/mL and 24 h were the optimal concentration and time of ROE. Compared with the blank control group, the expression of ALP protein in MC3T3-E1 cells was decreased (P<0.05), Runx2. The expression of BMP2 and Osteocalcin was significantly decreased (P<0.01). Compared with the model control group, the expression levels of ALP and BMP2 in the ROE administration group were increased (P<0.05), and the expression levels of Osteocalcin and Runx2 protein were significantly increased (P<0.01). Compared with the blank group, the model group ALP, Runx2 and BMP2 gene expressions were down-regulated (P<0.05). Osteocalcin was significantly down-regulated (P<0.01). Compared with the model group, ALP and Runx2 gene expressions were up-regulated in ROE-administered group (P<0.05). BMP2 and Osteocalcin gene expression were significantly down-regulated (P<0.01). Conclusion ROE can regulate the expression levels of bone metabolism genes and proteins and has a certain protective effect on the MC3T3-E1 senile osteoporosis model damaged by H2O2.