P物质上调正常人真皮成纤维细胞TGF-β_1的mRNA表达

Substance P up-regulates the TGF-β_1 mRNA expression of human dermal fibroblasts in vitro

目的 探讨神经肽P物质 (SubstanceP ,SP)与瘢痕形成间的调控关系。方法 体外分离培养正常人真皮成纤维细胞 ,分别加入SP及其受体特异性拮抗剂L 70 3,6 0 6乙酸盐 ,MTT法测定细胞生长增殖量 ;采用细胞爬片法培养细胞 ,加入SP对细胞作用后 ,应用TGF β1 mRNA特异性探针行细胞原位杂交 ,计算细胞的TGF β1 mRNA阳性表达率 ,并经图像分析测定阳性细胞TGF β1 mRNA的表达强度 ,分析SP与真皮成纤维细胞TGF β1 mRNA表达间的关系。结果 SP可促进体外培养的正常人真皮成纤维细胞的生长 ,其作用与SP浓度呈现依赖关系 ,当浓度达到 2 5ng ml时 ,刺激细胞生长的速率达最大值 ,可使细胞较对照组提前 4d融合 ;正常人真皮成纤维细胞受 2 5ng ml的SP刺激后TGF β1 mRNA的表达明显上调 ,4 8h后即可见细胞表达TGF β1 mRNA的阳性率及强度均显著增加 ;SP对正常人真皮成纤维细胞的上述效应 ,均可被其受体拮抗剂L 70 3,6 0 6乙酸盐所抑制。结论SP可促进正常人真皮成纤维细胞的生长增殖 ,同时上调细胞的TGF β1 mRNA表达 ;

Objective\ To investigate the role of substance P in the formation of hypertrophic scar. Methods\ Dermal fibroblasts derived from human normal skin were cultured with substance P alone or together with selective non peptide NK 1 tachykinin antagonist, L 703,606 oxalate salt. The effect of substance P on proliferation of fibroblasts was measured by MTT assay. Furthermore, the TGF β 1 mRNA expression in the fibroblasts was determined by in situ hybridization and image analysis. Results\ Substance P enhanced fibroblast proliferation dose dependently, which showed the maximum rate when the concentration of substance P was 25ng/ml or higher in the culture media. By 48 hours cultured with 25ng/ml of substance P, the fibroblasts expressed TGF β 1 mRNA more significantly than the fibroblasts without substance P. The effects of substance P on both fibroblast proliferation and TGF β 1 mRNA expression could be antagonized by L 703,606 oxalate salt. Conclusion The results suggest that substance P may play an important role in phenotype changes of fibroblasts in skin scarring.\;