P物质对体外培养大鼠成纤维细胞表皮生长因子及其受体基因表达的影响

Effect of substance P on EGF and EGFR gene expressions in cultured rat fibroblasts

目的 探讨P物质 (SubstanceP ,SP)作用于离体培养的大鼠肉芽组织成纤维细胞后 ,对其表皮生长因子(EGF)及受体 (EGFR)基因表达的影响。方法 采用大鼠肉芽组织成纤维细胞培养和逆转录 聚合酶链反应 (RT PCR)技术 ,观察SP在不同浓度 ( 1× 10 -9～ 1× 10 -5mol/L)及不同孵育时间 ( 0、3、6、12、2 4h)情况下刺激大鼠成纤维细胞后 ,成纤维细胞EGF、EGFRmRNA表达的改变情况。结果 SP可上调大鼠肉芽组织成纤维细胞EGF、EGFRmRNA表达。其中 ,SP对EGF的量 效曲线呈双相分布 ,最大效应浓度为 1× 10 -7mol/L。但SP仅在高浓度时 ( 1× 10 -6～ 1× 10 -5mol/L)促进EGFR表达。在最大效应浓度 ( 1× 10 -7mol/L ,1× 10 -5mol/L)时 ,SP对大鼠成纤维细胞EGF、EGFRmRNA表达的上调作用分别于孵育细胞后 6、12h达高峰。结论 SP对大鼠肉芽组织成纤维细胞EGF、EGFR基因表达存在直接影响 ,其表现形式与SP的刺激浓度及孵育时间有关。这种影响可能是SP在创伤愈合过程中发挥调控作用的生物学机制之一。

Objective To explore the effect of substance P(SP) on epidermal growth factor(EGF) and epidermal growth factor receptor(EGFR) gene expressions in cultured rat granulation tissue fibroblasts. Methods The changes of EGF and EGFR gene expressions in cultured rat granulation tissue fibroblasts treated with SP at different concentrations(1×10 -9 ～1×10 -5 mol/L) and incubating time(0, 3, 6, 12 and 24 h) were observed by semi quantitative RT PCR. Results SP could up regulate EGF and EGFR gene expressions in rat granulation tissue fibroblasts in vitro . The response curve of EGF treated by SP showed a two way distribution at the maximal effect concentration of 1×10 -7 mol/L, but SP could up regulate EGFR gene expression only at higher concentrations(1×10 -6 ～1×10 -5 mol/L). After stimulation of rat fibroblasts with SP at 1×10 -7 mol/L and 1×10 -5 mol/L, EGF and EGFR gene expressions peaked at 6 and 12 h, respectively. Conclusion SP has a direct effect on EGF and EGFR gene expressions in rat fibroblasts in vitro in dose and time dependent manners. This effect might be one of the biological mechanisms of the regulatory role of SP during wound healing.