增生性瘢痕成纤维细胞P物质表达的研究

目的研究P物质（substance P，SP）在人增生性瘢痕体外培养的成纤维细胞的表达情况及其与正常皮肤之间的差异，探讨增生性瘢痕组织SP表达增高的原因。方法以酶消化法体外培养成人增生性瘢痕以及正常皮肤的成纤维细胞，将SP（终浓度为10^-7mol／L）加入培养液中刺激成纤维细胞，分别于刺激前和刺激后1、3、6、12、24h采用RT—PCR检测细胞内SP基因的表达情况，ELISA检测培养液中SP浓度。正常皮肤对照组及瘢痕对照组除不应用SP外，余处理均相同。结果未受SP刺激时，人增生性瘢痕及正常皮肤体外培养的成纤维细胞均能分泌一定浓度的SP，两者之间差异有统计学意义（P〈0．01）。SP刺激后，正常皮肤体外培养的成纤维细胞SP基因和蛋白表达均在1～3h内达到高峰，随后迅速下降，24h后仍高于其正常水平；而增生性瘢痕体外培养的成纤维细胞SP基因和蛋白表达在3～6h内达到高峰，随后缓慢下降，24h后仍高于其正常水平。两种成纤维细胞SP的分泌差异有统计学意义（P〈0．05）。结论外源性SP可诱使成纤维细胞内SP的分泌增加，而增生性瘢痕与正常皮肤成纤维细胞SP的分泌情况有明显差异。

Expression of substance P in cultured hypertrophic scar fibroblasts

Objective To investigate the differential expression of substance P(SP)in normal and hypertrophic scar fibroblasts in vitro and the reason of its high expression in the latter.Methods The normal and hypertrophic scar fibroblasts were isolated by enzyme digestion and cultured.SP with a final concentration of 10-7 mol/L were added into the cell culture media,and those without SP stimulation served as control.Before and 1,3,6,12 and 24 h after SP stimulation,the SP expressions in the media of 4 groups were detected by RT-PCR and its concentrations was measured by ELISA.Results Without SP treatment,the fibroblasts from normal and hypertrophic scar excreted certain with significant difference between them(P<0.01).In presence of SP,the gene and protein expression of SP of normal fibroblasts reached a peak level in 3 to 6 h,and then decreased rapidly to normal level in 24 h.For the fibroblasts from hypertrophic scar,the SP showed similar tendency with normal group after SP treatment,but kept a high level in 24 h.There were significant difference between the normal fibroblasts and the ones from hypertrophic scar after SP treatment(P<0.01).Conclusion SP can induce the secretion of SP in fibroblasts,and SP secreted by normal and hypertrophic scar fibroblasts in vitro was statistically significant.