Role of transforming growth factor-β and extracellular regulated protein kinases in lipopolysaccharide-induced osteoblastic differentiation of porcine aortic valve interstitial cells

Hao FENG; Qiong SHI; Yingjiu JIANG

doi:10.16016/j.1000-5404.202001152

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Role of transforming growth factor-β and extracellular regulated protein kinases in lipopolysaccharide-induced osteoblastic differentiation of porcine aortic valve interstitial cells

Basic Medicine | 更新时间：2026-01-09

- Role of transforming growth factor-β and extracellular regulated protein kinases in lipopolysaccharide-induced osteoblastic differentiation of porcine aortic valve interstitial cells
- Journal of Third Military Medical University Vol. 42, Issue 10, Pages: 989-995(2020)
- 作者机构：
  
  1.400016 重庆，重庆医科大学：附属第一医院胸心外科
  2.400016 重庆，重庆医科大学：检验医学院
- 作者简介：
- 基金信息：
- DOI：10.16016/j.1000-5404.202001152
  CLC： R322.121;R329.21;R329.24
- Received：06 January 2020，
  
  Revised：2020-03-02，
  
  Published：30 May 2020
- 稿件说明：
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Hao FENG, Qiong SHI, Yingjiu JIANG. Role of transforming growth factor-β and extracellular regulated protein kinases in lipopolysaccharide-induced osteoblastic differentiation of porcine aortic valve interstitial cells[J]. Journal of Third Military Medical University, 2020, 42(10): 989-995.
DOI：

Hao FENG, Qiong SHI, Yingjiu JIANG. Role of transforming growth factor-β and extracellular regulated protein kinases in lipopolysaccharide-induced osteoblastic differentiation of porcine aortic valve interstitial cells[J]. Journal of Third Military Medical University, 2020, 42(10): 989-995. DOI： 10.16016/j.1000-5404.202001152.

摘要

目的

探讨转化生长因子-β1(transforming growth factor-β，TGF-β1)、细胞外调节蛋白激酶(extracellular regulated protein kinases，ERK)在脂多糖(lipopolysaccharide，LPS)诱导猪主动脉瓣膜间质细胞(porcine aortic valve interstitial cells，PVICs)成骨分化过程中的作用与机制。

方法

体外分离并培养PVICs。应用LPS诱导PVICs成骨分化，ALP染色和茜素红S染色检测细胞成骨分化能力；RT-qPCR法检测细胞成骨相关基因及蛋白Runt相关转录因子2 (runt-related transcription factor 2，Runx2)、骨桥蛋白(osteopontin，OPN)和骨钙蛋白(osteocalcin，OCN)的转录水平。Western blot法检测TGF-β1、TGF-βR、Bax、Smad2/3和ERK1/2的蛋白表达水平。同时应用ERK特异性抑制剂PD98059处理，观察ERK、Bax、Runx2蛋白表达水平。

结果

① LPS(4 μg/mL)处理可以使PVICs的碱性磷酸酶活性明显升高，钙盐沉积明显增强。②RT-qPCR检测结果显示：LPS(4 μg/mL)处理后可以明显上调Runx2、OPN和OCN的mRNA转录水平(

0.01)。③Western blot检测结果显示：LPS(4 μg/mL)处理后TGF-β1、TGF-βR、Smad2/3及ERK1/2蛋白水平显著升高(

0.05)；ERK特异性抑制剂PD98059可以减弱LPS对ERK、Bax、Runx2的促表达作用(

0.05)。

结论

LPS可能通过激活TGF-β1/Smads和ERK信号通路诱导主动脉瓣膜间质细胞成骨分化

Abstract

Objective

To investigate the role of transforming growth factor-β (TGF-β1) and extracellular regulated protein kinases (ERK) in lipopolysaccharide (LPS)-induced osteoblastic differentiation of porcine aortic valve interstitial cells (PVICs).

Methods

In vitro

cultured PVICs were treated with LPS (4 μg/mL) to induce their osteogenic differentiation

and early and late oste

ogenic differentiation abilities of the cells were assessed using alkaline phosphatase (ALP) staining and Alizarin red S staining. The expression levels of Runx2

OPN and OCN mRNAs in the cells were detected using RT-qPCR

and those of TGF-β1

TGF-βR

Bax

Smad2/3 and ERK1/2 proteins were determined using Western blotting.

Results

In cultured PVICs

LPS treatment significantly increased alkaline phosphatase activity

deposition of calcium salts

and the expression levels of Runx2

OPN and OCN mRNA (

0.01). In addition to the increase of the expression of osteoblastic differentiation markers

the protein expression of TGF-β1

TGF-βR

Smad2/3 and ERK1/2 were also significantly increased in LPS-treated PVICs (

0.05); treatment with PD98059

a specific ERK1/2 inhibitor

significantly abolished LPS-induced increase in the protein expression of ERK1/2

Bax

and Runx2 (

0.05).

Conclusion

LPS induces osteoblastic differentiation of PVICs probably by activating TGF-β1/Smads and ERK signaling pathways.

关键词

Keywords

references

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