碱性成纤维细胞生长因子在P物质诱导肉芽组织成纤维细胞增殖中的作用

Effects of basic fibroblast growth factor on proliferation of granulation tissue fibroblasts induced by substance P

目的 探讨感觉神经肽P物质 (substanceP ,SP)对碱性成纤维细胞生长因子 (basicfibroblastgrowthfactor ,bFGF)表达的调控作用及其在SP促离体培养的肉芽组织成纤维细胞增殖中的作用。方法 采用MTT法测定SP对原代培养的肉芽组织成纤维细胞的促增殖作用 ;采用RT PCR和Westernblotting方法检测SP对成纤维细胞bFGF基因和蛋白表达的调控作用 ,观察时间及剂量 效应关系。结果 10 -9～ 10 -5mol/L的SP在体外对原代培养的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) ,bFGF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bFGFmRNA的表达 ,在作用后 3 ,6h与对照组相比 ,差异有非常显著性意义 (P <0 0 1) ;12h后可检测到bFGF蛋白明显表达增强 (P <0 0 1) ,2 4h达高峰 ,48h后逐渐有所回落。SP在 10 -9～ 10 -5mol/L范围内可显著促进成纤维细胞bFGFmRNA表达 ,在 10 -8～ 10 -5mol/L范围可诱导bFGF蛋白的表达 ,均在 10 -7mol/L剂量点达到峰值 (P <0 0 1)。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 regulatory effect of sensory neuropeptide substance P (SP) on the expression of basic fibroblast growth factor (bFGF) and the proliferation promoting effect of bFGF on the in vitro cultured granulation tissue fibroblasts induced by SP. Methods The proliferation promoting effect of SP on the cultured granulation tissue fibroblasts was observed by MTT. The regulatory effect of SP on the expression of fibroblast bFGF was determined by RT PCR and Western blotting. The time and dose effect relations were also observed. Results SP had obvious proliferation promoting effect on the primary cultured granulation tissue fibroblasts in vitro in a remarkable dose dependent manner ( r =0.594, P <0 01). However, bFGF antibody could partly exert its inhibitory effect. The expression of bFGF mRNA was more significant at 3 and 6 h after induction of SP at 10 -7 mol/L, as compared with that of the control group ( P <0 01). The expression of bFGF protein was markedly higher than that of the control group at 12 h, reached the peak at 24 h, and declined gradually at 48 h. SP at the concentrations of 10 -9 -10 -5 mol/L could significantly promote the expression of bFGF mRNA and that at 10 -8 -10 -5 mol/L could induce the expression of bFGF protein. Both the expression levels of bFGF mRNA and bFGF protein reached the peak when the concentration of SP was at 10 -7 mol/L ( P <0.01). Conclusion SP has a significant proliferation promoting effect on the granulation tissue fibroblasts, which is correlated with bFGF mRNA expression of fibroblasts induced by SP.