神经肽P物质对肉芽组织成纤维细胞增殖及表皮生长因子基因表达的作用

Effects of sensory neuropeptide substance P on cellular proliferation of granulation tissue fibroblast and gene expression of epidermal growth factor mRNA

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的促增殖作用及其对表皮生长因子 (EGF)基因表达的调控作用。 方法 采用MTT法测定SP对肉芽组织成纤维细胞的促增殖作用 ;采用逆转录 -聚合酶链反应 (RT -PCR)方法检测SP对成纤维细胞EGF基因表达的调控作用 ,观察时间及剂量 -效应关系。 结果 10 - 9～ 10 - 5mol L的SP在体外对肉芽组织成纤维细胞均具有明显的促增殖作用 (P <0 .0 1) ,且具有明显的剂量依赖性 (r =0 .5 94,P <0 .0 1) ,EGF抗体只能部分抑制这一作用 (与对照组比较 ,P <0 .0 1;与 10 - 7mol LSP组比较 ,P <0 .0 5 )。 10 - 7mol LSP可诱导成纤维细胞EGFmRNA的表达 ,在作用后 6h与对照组比较 ,差异有非常显著性意义 (P <0 .0 1) ;SP在 10 - 8～ 10 - 6 mol L范围内可以显著促进成纤维细胞EGFmRNA表达 ,在 10 - 7mol L达到峰值 ,当浓度 >10 - 7mol L时 ,其促EGFmRNA表达的效应强度随浓度升高而有所降低 ,至 10 - 5mol L时与对照组比较 ,差异无显著性意义。 结论 SP对肉芽组织成纤维细胞具有明显的促增殖作用 。

Objective To explore the proliferation promoting effect of sensory neuropeptide substance P (SP) on the in vitro cultured granulation tissues fibroblasts and its regulative effect on the gene expression of epidermal growth factor (EGF) mRNA. Methods The proliferation promoting effect of cultured granulation tissue fibroblasts was observed by means of MTT, the regulative effect of SP on gene expression of EGF in fibroblasts by RT PCR. The time and dose efficiency relations were also observed. Results There was a significant proliferation promoting effect of SP at concentration of 10 9 10 5 mol/L on the cultured granulation tissue fibroblasts in vitro in a remarkable dose dependent fashion ( r =0.594, P <0.01). However, EGF antibody only partly exerted its inhibitive effect ( P <0.01 compared with control group; P <0.05 compared with Group SP at 10 7 mol/L). SP at 10 7 mol/L could induce the EGF mRNA expression of the fibroblasts with a very significant difference after 6 hours compared with control group ( P <0.01). SP could promote the EGF mRNA expressions of the fibroblasts at the concentration of 10 8 10 6 mol/L and reached peak at 10 7 mol/L. When the concentration was over 10 7 mol/L, the promoting effect of SP on the EGF mRNA expression began to decrease in response to the increase of the concentration. There was no significant difference compared with control group when the concentration was up to 10 5 mol/L. Conclusions SP has a significant proliferation promoting effect on the granulation tissue fibroblasts, which is correlated with SP inducing EGF mRNA expression of fibroblasts.