Role of FSHβ in BMP9-induced osteogenic differentiation in mouse embryonic fibroblasts

Su Xiaoya; Huang Min; Liu Hong

doi:10.13406/j.cnki.cyxb.001200

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Role of FSHβ in BMP9-induced osteogenic differentiation in mouse embryonic fibroblasts

更新时间：2024-01-17

- Role of FSHβ in BMP9-induced osteogenic differentiation in mouse embryonic fibroblasts
- Journal of Chongqing Medical University Vol. 42, Issue 3, Pages: 327-333(2017)
- 作者机构：
  
  重庆医科大学附属第一医院生殖健康与不孕症中心
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.001200
  CLC： R580
- Published：2017，
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Su Xiaoya, Huang Min, Liu Hong. Role of FSHβ in BMP9-induced osteogenic differentiation in mouse embryonic fibroblasts. [J]. Journal of Chongqing Medical University 42(3):327-333(2017)
DOI：

Su Xiaoya, Huang Min, Liu Hong. Role of FSHβ in BMP9-induced osteogenic differentiation in mouse embryonic fibroblasts. [J]. Journal of Chongqing Medical University 42(3):327-333(2017) DOI： 10.13406/j.cnki.cyxb.001200.

摘要

目的:探讨卵泡刺激素β亚基(follicle-stimulating hormoneβ

FSHβ)对骨形态发生蛋白9(bone morphogenetic protein 9

BMP9)诱导小鼠胚胎成纤维细胞(mouse embryonic fibroblasts

MEFs)成骨分化中的作用。方法:采用Ad Easy系统构建表达绿色荧光蛋白(green fluorescent protein

GFP)、FSHβ和BMP9的重组腺病毒(Ad-GFP、Ad-FSHβ和Ad-BMP9)

实验分组为:AdGFP组、Ad-BMP9组、Ad-FSHβ组和Ad-BMP9+Ad-FSHβ组。运用化学发光定量和组织化学染色法分别检测各组3、5、7 d碱性磷酸酶(alkaline phosphatase

ALP)活性;Western blot检测成骨分化早期转录因子(Runx2和DLX5)的蛋白表达水平;RT-PCR检测成骨分化晚期指标骨钙蛋白(osteocalcin

OCN)和骨桥蛋白(osteopontin

OPN)的m RNA表达水平;茜素红染色检测钙盐沉积;流式细胞技术检测各实验组细胞的周期分布。结果:在MEFs中

过表达FSHβ能明显促进BMP9诱导MEFs骨化过程中第3天(6 874.67±224.93 vs.11 935.67±568.76

P=0.000)、第5天(43 434.67±2 047.57 vs.98 913.33±4 606.26

P=0.000)、第7天(38 198.00±2 376.43 vs.78 534.67±2 114.56

P=0.000)的ALP活性

转录因子Runx2(1.171±0.105 vs.1.350±0.053

P=0.026)、DLX5(1.382±0.123 vs.1.738±0.286

P=0.001)和晚期成骨指标OCN(1.300±0.045 vs.1.471±0.046

P=0.002)、OPN(1.300±0.084 vs.1.479±0.038

P=0.016)的表达水平

以及钙盐沉积。此外

FSHβ和(或)BMP9对MEFs细胞周期没有明显影响(G1期:F=0.226

P=0.876;G2期:F=0.147

P=0.929;S期:F=0.027

P=0.994)。结论:在不影响MEFs细胞周期的情况下

FSHβ协同BMP9诱导MEFs成骨分化。

Abstract

Objective:To study the effect of follicle-stimulating hormone β（FSHβ） on bone morphogenetic protein 9（BMP9）-induced osteogenic differentiation in mouse embryonic fibroblasts（MEFs）. Methods:Ad Easy system was used for generating target recombinant adenoviruses（Ad-GFP

Ad-FSHβ

and Ad-BMP9）

and our investigation was designed into four groups:Ad-GFP group

Ad-BMP9 group

Ad-FSHβ group

and Ad-BMP9+Ad-FSHβ group. Chemiluminescence technique and histochemical staining were conducted to assay alkaline phosphatase（ALP）activity on day 3

and 7

Western blot to test the protein expression of Runx2 and Dlx5

RT-PCR to measure the m RNA expression of osteocalcin（OCN）and osteopontin（OPN）

and Alizarin Red S staining to detect the matrix mineralization. In addition

Flow cytometry was utilized for analysing the cell cycle of different groups. Results:Exogenous expression of FSHβ dramatically enhanced the ALP activity on day 3（6 874.67±224.93 vs. 11 935.67±568.76

P=0.000）

day 5（43 434.67±2 047.57 vs. 98 913.33±4 606.26

P=0.000）

and day 7（38 198.00±2 376.43 vs. 78 534.67±2 114.56

P=0.000）

and the protein expression of Runx2（1.171±0.105 vs. 1.350±0.053

P=0.026）and DLX-5（1.382±0.123 vs. 1.738±0.286

P=0.001）

and the m RNA expression of late osteogenic makers

namely OCN（1.300±0.045 vs. 1.471±0.046

P=0.002）and OPN（1.300±0.084 vs. 1.479±0.038

P=0.016）

and matrix mineralization induced by BMP9 in MEFs. Moreover

efficient infection of AdFSHβ or/and Ad-BMP9 remains no apparent influence on cell cycle in MEFs（G1-phase:F =0.226

P =0.876;G2-phase:F =0.147

P=0.929;S-phase:F=0.027

P=0.994）. Conclusion:FSHβ promotes BMP9-induced osteogenic differentiation without substantial effect on cell cycle in MEFs.

关键词

骨形态发生蛋白9卵泡刺激素β亚基小鼠胚胎成纤维细胞成骨分化

Keywords

bone morphogenetic protein 9follicle-stimulating hormone βmouse embryonic fibroblastsosteogenic differentiation

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