信号传导及转录激活因子1基因沉默促进人牙周膜细胞成骨细胞分化

摘要/Abstract

摘要： ［摘要］　目的　探讨信号传导及转录激活因子1（signal transducers and activators of transcription 1，STAT1）对人牙周膜细胞骨向分化的作用及其机制。方法　采用酶消化法分离培养原代人牙周膜细胞。采用细菌脂多糖（Lipopolysaccharide，LPS）刺激人牙周膜细胞。采用成骨分化培养基诱导人牙周膜细胞骨向分化。通过转染STAT1过表达载体或小干扰RNA（small interfering RNA，siRNA）实现STAT1基因过表达或基因表达沉默。采用碱性磷酸酶（alkaline phosphatase，ALP）活性检测试剂盒和茜素红染色检测成骨分化。实时定量PCR方法检测STAT1，I型胶原蛋白（collagen type Iα1，ColA1）和骨调素（osteopontin，OPN）的mRNA表达水平。Western blot方法检测STAT1和Runt相关转录因子2（runt-related transcription factor 2，Runx2）的蛋白表达水平。结果　LPS炎性刺激诱导人牙周膜细胞STAT1高表达。STAT1过表达显著抑制ALP活性、骨质矿化、Runx2的核转移以及ColA1和OPN的表达水平。相反，STAT1基因沉默则显著促进ALP活性、骨质矿化、Runx2的核转移以及ColA1和OPN的表达水平。结论　STAT1基因沉默促进人牙周膜细胞成骨分化，其机制与促进Runx2核转移有关。

关键词: 信号传导及转录激活因子1, 人牙周膜细胞, 成骨细胞分化

Abstract: Abstract: Objective To investigate the role of signal transducers and activators of transcription 1 (STAT1) on osteoblast differentiation of human periodontal ligament cells and the underlying mechanism. Methods Primary human periodontal ligament cells were isolated and cultured by enzyme-digestion method. Lipopolysaccharide (LPS) was used to stimulate human periodontal ligament cells. The osteoblast differentiation of human periodontal ligament cells was induced by osteogenetic differentiation medium. STAT1 overexpression or gene silencing was achieved by transfection of STAT1 overexpression vectors or small interfering RNA (siRNA), respectively. Osteoblast differentiation was detected by alkaline phosphatase (ALP) assay kit and Alizarin red S staining. The mRNA expression levels of STAT1, collagen type Iα1 (ColA1) and osteopontin (OPN) were detected by real-time quantitative PCR analysis. The protein expression levels of STAT1 and runt-related transcription factor 2 (Runx2) were detected by Western blot analysis. Results LPS stimulation significantly upregulated the expression of STAT1. Overexpression of STAT1 significantly inhibited the ALP activity, bone matrix mineralization, nuclear translocation of Runx2 and the expression of ColA1and OPN. In contrast, STAT1 gene silencing markedly increased the ALP activity, bone matrix mineralization, nuclear translocation of Runx2 and the expression of ColA1and OPN. Conclusions STAT1 gene silencing promotes osteoblast differentiation of human periodontal ligament cells through increasing the nuclear translocation of Runx2.

Key words: signal transducers and activators of transcription 1, human periodontal ligament cells, osteoblast differentiation

中图分类号:

呼海燕刘彩宏. 信号传导及转录激活因子1基因沉默促进人牙周膜细胞成骨细胞分化[J]. , 2016, 36(10): 884-889.

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