核因子I-C降低皮肤成纤维细胞对TGF-β的敏感性

Nuclear factor I-C inhibits platelet-derived growth factor-induced enhancement of dermal fibroblast sensitivity to TGF-β

目的探讨核因子I-C(NFI-C)对血小板源性生长因子(PDGF)促进皮肤成纤维细胞表达TGF-β受体Ⅱ(TβRⅡ)作用的影响。方法含有NFI-C序列的慢病毒转染人皮肤成纤维细胞(HFF-1);根据细胞生长活性及转染效率筛选最佳转染复数(MOI);PDGF-BB刺激体外培养的HFF-1细胞、转染NFI-C的HFF-1细胞以及转染阴性病毒的HFF-1细胞,并设立转染NFI-C但不用PDGF处理的细胞;以不做任何处理的HFF-1细胞作为空白对照组;采用western blot、RT-q PCR测定各组细胞TβRⅡ的表达。数据采用单因素方差分析、LSD-t及SNK-q检验对各组数据进行分析。结果慢病毒转染HFF-1最适MOI为50。PDGF处理的HFF-1细胞表达TβRⅡ高于空白对照组(P<0.05);在PDGF作用下,转染NFI-C的HFF-1细胞表达TβRⅡ低于未转染的细胞(P<0.05);阴性病毒无明显抑制作用(P>0.05);单纯转染NFI-C的HFF-1细胞表达TβRⅡ与空白对照组无显著性差异(P>0.05)。结论NFI-C能抑制PDGF对TβRⅡ的上调作用,降低皮肤成纤维细胞对TGF-β的敏感性。

Objective To investigate the effect of nuclear factor I- C（NFI- C） on platelet- derived growth factor（PDGF）- induced up-regulation of TGF-β receptor II（TβRII） in dermal fibroblasts. Methods A lentiviral vector containing NFI-C sequence（LentiGFP- NFI- C） was transfected into a human foreskin fibroblast cell line（HFF- 1）. Cultured HFF- 1 cells, cells transfected with Lenti- GFP- NFI- C, and cells transfected with a negative virus were stimulated with PDGF- BB, and Western blotting and RTq PCR were used to detect the expression levels of TβRII in the treated cells. Results PDGF treatment significantly increased the expression level of TβRII in HFF- 1 cells（P〈0.05）. The cells transfected with Lenti- GFP- NFI- C expressed a significantly lower level of TβRII than non- transfected cells in response to PDGF stimulation（P〈0.05）, but the negative virus showed no such inhibitory effect（P〈0.05）. No significant difference was found in the expression level of TβRII protein between cells transfected with Lenti-GFP-NFI-C-transfection before PDGF stimulation and the blank control cells. Conclusion NFI-C can inhibit PDGFinduced up-regulation of TβRII and thus reduce the sensitivity of the dermal fibroblasts to TGF-β.