中国骨质疏松杂志

通过p38MAPK信号通路探讨仙灵骨葆胶囊对MC3T3-E1成骨分化的影响

The effect of Xianling Gubao capsule on osteogenic differentiation of MC3T3-E1 through p38 MAPK signaling pathway

DOI：10.3969/j.issn.1006-7108.2022.06.016

中文关键词: 仙灵骨葆胶囊 MC3T3-E1 p38MAPK信号通路骨代谢分子机制

英文关键词:Xianling Gubao capsules MC3T3-E1 p38MAPK signaling pathway bone metabolism molecular mechanism

基金项目:甘肃省自然科学基金项目（17JR5RA056）

作者	单位
张岩董万涛* 安文博张碧峰	甘肃中医药大学附属医院，兰州 730020

摘要点击次数: 503

全文下载次数: 0

中文摘要:

目的探讨仙灵骨葆胶囊含药血清对小鼠胚胎成骨细胞前体细胞（MC3T3-E1）增殖分化的影响及与p38MAPK信号通路的关系。方法用仙灵骨葆胶囊含药血清干预MC3T3-E1，CCK8法筛选最佳干预时间及浓度，试剂盒检测各组细胞ALP活性。将细胞分为A、B、C、D 4组，分别加入10%含药血清、10%空白血清、10%含药血清+SB203580、10%空白血清+SB203580，利用免疫印迹法（Western blot）检测p38丝裂原活化蛋白激酶（p38）、磷酸化p38（p-p38）、成骨相关转录因子2（Runx2）、骨形态发生蛋白2（BMP-2）的蛋白表达量，荧光定量PCR检测p38、Runx2和BMP-2 mRNA的表达水平。结果与空白血清组相比，不同浓度的仙灵骨葆胶囊含药血清均能促进MC3T3-E1的增殖分化，提高ALP的活性，其中10%含药血清干预36 h的作用最为明显（P＜0.05）；与B组比较，A组p38、p-p38、Runx2、BMP-2蛋白及p38、Runx2、BMP-2 mRNA的表达明显增高（P＜0.05）；加入信号通路阻断剂SB203580后，C组与D组上述指标的表达明显降低（P＜0.05）；与C组相比，D组p38、p-p38、Runx2、BMP-2蛋白及p38、Runx2、BMP-2 mRNA表达更低（P＜0.05）。结论仙灵骨葆胶囊含药血清能促进MC3T3-E1的分化生长，其作用机制可能与激活p38MAPK信号通路、上调成骨相关因子Runx2、BMP-2的表达有关。

英文摘要:

Objective To investigate the effect of conditioned serum of Xianling Gubao capsules on the proliferation and differentiation of mouse embryonic osteoblast precursor cells (MC3T3-E1) and its relationship with p38 MAPK signaling pathway. Methods MC3T3-E1 cells were intervened with conditioned serum of Xianling Gubao capsules. The optimal intervention time and concentration were selected using CCK8 assay. ALP activity of cells in various groups was detected with kit. The cells were divided into four groups: A, B, C, and D. They were conditioned with 10% conditioned serum, 10% blank serum, 10% conditioned serum+SB203580, and 10% blank serum+SB203580, respectively. The protein expressions of p38 mitogen-activated protein kinase (p38), phosphorylated p38 (p-p38), osteogenesis-related transcription factor 2 (Runx2), and bone morphogenetic protein 2 (BMP-2) were detected using immunoblotting (Western blotting). The mRNA expressions of p38, Runx2, and BMP-2 were detected with real-time fluorescence quantitative PCR. Results Compared with the blank serum group, conditioned serum at different concentrations of Xianling Gubao capsules promoted the proliferation and differentiation of MC3T3-E1 cells and increased the activity of ALP. The most significant effect was achieved at 10% conditioned serum intervening for 36h (P<0.05). Compared to those in group B, the expression levels of p38, p-p38, Runx2, and BMP-2 protein and p38, Runx2, BMP-2 mRNA were significantly increased in group A (P<0.05). After adding SB203580, a blocker of signaling pathway, the expressions of the above indicators were significantly lower in the C and D groups (P<0.05). The expression of p38, p-p38, Runx2 and BMP-2 protein and p38, Runx2, BMP-2 mRNA in group C were lower than those in group D (P<0.05). Conclusion Conditioned serum of Xianling Gubao capsules promotes the differentiation and growth of MC3T3-E1 cells. Its mechanism may be related to the activation of p38MAPK signaling pathway and the up-regulation of the expressions of osteogenesis-related factors Runx2 and BMP-2.

查看全文查看/发表评论下载PDF阅读器

关闭