核因子-κB诱导激酶和IκB激酶在碱性成纤维细胞生长因子作用下基因表达及肌腱细胞增殖

Gene expression and tenocyte proliferation of nuclear factor-κB under the stimulation of basic fibroblast growth factor

目的 探讨核因子 -κB(NF -κB)、NF -κB诱导激酶 (NIK)和其抑制蛋白 (IκB)激酶(IKK)在碱性成纤维细胞生长因子 (bFGF)促肌腱细胞NF -κB基因表达信号传递中的作用。 方法 用兔的趾屈肌腱细胞 ,分成三组 ,分别在无bFGF、2ng mlbFGF和 10ng mlbFGF环境下培养5d ,绘制肌腱细胞生长曲线 ,5d时提取mRNA ,逆转录 -聚合酶链反应 (RT -PCR)检测NIK、IκB激酶α(IKKα)、IκB激酶 β(IKKβ)基因表达水平。 结果 随使用bFGF剂量增加 ,肌腱细胞生长曲线前移。 2ng mlbFGF组的NF -κB、NIK、IKKα、IKKβ基因表达相对值分别为 0 .4± 0 .2 ,0 .4± 0 .1,0 .3± 0 .1,0 .2± 0 .1;10ng mlbFGF组各基因分别为 1.8± 0 .5 ,1.0± 0 .3,0 .8± 0 .2 ,1.5± 0 .4 ,bFGF加入后 4种基因表达均显著增加 (P <0 .0 1)。 结论 bFGF增强NF -κB通路上各信号传递因子的基因表达并促进肌腱细胞增殖 ,提示bFGF促进腱细胞增殖的信号传递系统可能为NF -κB系统。

Objective To investigate the effect of nuclear factor-κB (NF-κB), NF-κB-inducing kinase (NIK) and I-κB in basic fibroblast growth factor (bFGF) promoting gene expression and signal transduction of tenocyte NF-κB. Methods Tenocytes isolated from rabbits were divided into 3 groups with 8 dishes: no bFGF (Group A), 2 ng/ml bFGF (Group B) and 10 ng/ml bFGF (Group C). Cells in all dishes were cultured for 5 days. The effect of bFGF on tenocytes proliferation was examined by means of growth curve analysis. The mRNA was isolated after 5 days and the gene expressions of NIK,IKKα and IKKβ induced by NF-κB investigated by reverse transcription-polymerase chain reaction (RT-PCR). Results The growth curve moved forward with the dosage increase of bFGF. Gene expressions of NF-κB,NIK, IκB kinase (IKK)α and IKKβ were 0.4± 0.2 , 0.4 ± 0.1 , 0.3 ±0.1 and 0.2±0.1 respectively in the Group B and 1.8±0.5,1.0±0.3, 0.8 ± 0.2 and 1.5 ± 0.4 in the Group C. Gene expressions were all up-regulated in the Groups B and C posterior to adding of bFGF. Conclusions bFGF stimulates tenocyte proliferation and up-regulates gene expression of NF-κB,NIK,IKKα and IKKβ, as indicates that the signal transduction of promoting tenocyte proliferation by bFGF may be through NF-κB pathway.