P物质对肉芽组织成纤维细胞表皮生长因子的表达调控及其意义

目的 探讨感觉神经肽P物质(SP)对离体培养的肉芽组织成纤维细胞表皮生长因子(EGF)表达的调控作用及特点。方法 采用甲醛注射的方法造成Wistar大鼠局部无菌性炎性反应，提取肉芽组织进行成纤维细胞原代培养；采用逆转录—聚合酶链反应(RT—PCR)方法检测SP对肉芽组织成纤维细胞EGF基因表达的调控作用，观察时间及剂量—效应关系；采用Western-blotting方法检测EGF蛋白表达情况，观察时间及剂量—效应关系。结果 10^-7mol/L SP可诱导成纤维细胞EGF mRNA的表达，在作用后6h与对照组比，差异有显著性(P＜0．01)；SP在10^-8-10^-6mol/L范围内可以显著促进成纤维细胞EGF mRNA表达，在10^-7mol/L达到峰值，10^-5mol/L时与对照组差异无显著性(P＞0．05)。10^-7mol/L SP作用12h后可检测到EGF蛋白明显表达增强(P＜0．01)，24h达高峰，48h后逐渐有所回落。SP在在10^-8--10^-5mol/L范围可诱导EGF蛋白的表达，均在10^-7mol/L剂量点达到峰值(P＜0．01)。结论 SP可诱导肉芽组织成纤维细胞EGF基因和蛋白的表达，且呈现出一定的时间和剂量特点，在SP调控创伤愈合的作用中具有重要意义。

Regulative effects of substance P on expression of epidermal growth factor of granulation tissue fibroblasts in vitro

Objective To explore the regulative effects and significance of neuropeptide substance P (SP) on the expression of epidermal growth factor (EGF) of granulation tissue fibroblasts in vitro.Methods A local asepsis inflammation was made by injection of formaldehyde in rats and its granulation tissue kept under culture.RT-PCR was employed to detect the expression of EGF mRNA after inducement of SP at different concentrations and time points in the granulation tissue fibroblasts and western blot to detect the expression of EGF protein.Results SP could induce the EGF mRNA expression of the fibroblasts,peaked at the 6th h and then decreased gradually.SP could promote the EGFmRNA expression of the fibroblasts in the concentration of 10 2?8 to 10 -6 mol/L,peaked in the concentration of 10 -7 mol/L.There was no significance in comparision with the control group at the concentration of 10 -5 mol/L.The expression level of EGF protein was markedly higher than the control group after 12 h,reached the peak at the 24th h and declined gradually after 48 h.SP at concentrations of 10 -8 to 10 -5 mol/L could significantly promote the expression of EGF protein,reached the peak when SP concentration was at 10 -7 mol/L ( P <0.01).Conclusion SP can induce the expression of EGF mRNA and EGF protein of granulation tissue fibroblasts in vitro,which may possess an important significance in wound healing.