脂多糖刺激后人正常皮肤成纤维细胞基因表达谱的变化及其意义

Changes in gene expression profiles of normal skin fibroblasts challenged by lipopolysaccharide and their significance

目的:观察正常皮肤成纤维细胞经大肠杆菌内毒素(LPS)刺激后基因表达谱的变化,探讨LPS对后期瘢痕形成的影响及可能机制。方法:用0.1μg/ml LPS刺激正常皮肤成纤维细胞并进行连续传代培养,选择第3代成纤维细胞,采用基因芯片技术检测成纤维细胞基因表达谱的变化,与自身正常皮肤成纤维细胞及自身增生性瘢痕组织成纤维细胞相关基因进行对比,挑选差异基因,采用逆转录-聚合酶链式反应(RT-PCR)方法进行验证。结果:LPS刺激后正常皮肤成纤维细胞基因表达谱发生改变,其中与胶原代谢相关的基因(Ⅰ型胶原、c-myc、TGF-β1mRNA)表达均上调;RT-PCR结果显示,这些基因表达与自身增生性瘢痕组织成纤维细胞表达量近似(P>0.05)。结论:LPS可能诱导正常皮肤成纤维细胞转化为增生性瘢痕组织成纤维细胞,参与增生性瘢痕形成。

Objective： To observe gene expression profiles of normal skin fibroblasts challenged by lipopolysaccharide （LPS）, and investigate its significance on hypertrophic scar formation. Methods： Twenty patients with hypertrophic scar in proliferative stage were selected randomly, primary cultured fibroblasts were obtained from their hypertrophic scar tissue and normal skin. Normal skin fibroblasts of passage 3 were challenged with 0. 1 μg/ml LPS. The gene expression profiles of fibroblasts were detected by cDNA microarray, and differential genes related to the formation of hypertrophic scar in LPS challenged normal skin fibroblasts and non-challenged normal skin fibroblasts, and they were validated by RT-PCR analysis. Hypertrophic scar fibroblasts of the same passage served as positive control group. Results： Gene expression profiles of normal skin fibroblasts changed after being challenged by 0. 1 μg/ml LPS, the expression of the procollagen I , c-myc and TGF-βI which were related to the colla- gen metabolism were up-regulated, the expressions of procollagen I , c-myc and TGF-β1 mRNAs were similar to those of the hypertrophic scar fibroblasts （P〉 0. 05）. Conclusions： The results suggest that LPS might convert normal skin fibroblasts to hypertrophic scar tissue fibrolasts and participate in the formation of hypertrophic scar.