阿魏酸钠对体外培养人真皮中成纤维细胞EGF表达的影响

Effect of Sodium Ferulate on Epidermal Growth Factor in Human Dermal Fibroblasts in Vitro

目的观察中药当归活性成分阿魏酸钠(sodium ferulate,SF)对人真皮成纤维细胞(fibroblast,FB)中表皮生长因子(epidermal growth factor,EGF)表达的影响。方法体外培养人真皮FB,用MTT法检测10μmol/L,30μmol/L,60μmol/L,125μmol/L,250μmol/L,500μmol/L和1 000μmol/L等不同浓度SF对FB增殖的影响;并用ELISA法分别重点观察SF 125μmol/L,250μmol/L和500μmol/L于24h,48h和72h时细胞上清液中EGF含量(pg/mg protein)。结果 10μmol/L,30μmol/L,60μmol/L,125μmol/L,250μmol/L和500μmol/L等不同浓度SF对FB增殖无明显影响(P>0.05),1 000μmol/L时细胞增殖显著性降低(P<0.05)。125μmol/L组24h,48h与72h EGF含量分别为548.23±137.28,589.25±120.28和654.28±132.10;250μmol/L组24h,48h和72h分别为589.65±157.29,701.27±193.47,745.23±129.43;500μmol/L组24h,48h和72h分别为757.24±111.47,903.24±134.12和950.76±129.28。125,250μmol/L组72h与24h比较明显升高(P<0.05),与48h比较无明显差异(P>0.05);500μmol/L组72h与48h比较明显升高(P<0.05),与24h比较显著性升高(P<0.01)。此外,125μmol/L和250μmol/L组24h与对照组比较无明显差异(P>0.05),500μmol/L组则显著性升高(P<0.01)。125μmol/L组48h与对照组比较无明显差异(P>0.05),250μmol/L,500μmol/L组较对照组显著性升高(P<0.01)。125μmol/L,250μmol/L和500μmol/L组72h时较对照组均显著性升高(P<0.05,P<0.01,P<0.01)。对照组不同时间比较无明显差异(P>0.05)。结论 当归活性成分SF能增强人体真皮成纤维细胞EGF的表达,且具有显著的剂量依赖性及时间依赖性。

Objective To investigate the effect of sodium ferulate on expression of epidermal growth factor （EGF） in culture of human dermal fibroblasts in vitro. Methods Human dermal fibroblasts were cultured artificially. After the human dermal fibroblasts were cultured by different concentration （10μmol/L,30μmol/L,60μmol/ L, 125 μmol/L, 250 μmol/L,500 μmol/L, 1 000 μmol/L ） of sodium ferulate for 72 hours, the cell multiplications were measured by MTY assay. The ceils were randomly divided into the following four groups： 125 μmol/L, 2501μmol/L,500 μmol/L and control groups. EGF level （ pg,/mg protein） which in cultural supernatants（CS）was measured by ELISA, after 24h, 48h and 72 hours respectively. Results Concentration of 10 μmol/L,301μmol/L, 60 μmol/L, 125 μmol/L, 250 μmol/L, 500 μmol/L of sodium ferulate groups had no effect for the proliferation of fibroblasts, but in 1 000μmol/L group it was significantly decreased on 1 000 Iμmol/L（P 〈0.01 ）. EGF level were 548.23 ± 137.28,589.25 ± 120.28,654.28 ± 132.10 at 24, 48 and 72h in 1251μmol/L group respectively. They were 589.65 ± 157.29,701.27 ± 193.47,745.23 ± 129.43 at 24, 48 and 72h in 250μmol/L group ; and 757.24 ± 111.47,903.24 ± 134.12,950.76 ± 129.28 at 24,48 and 72h in 5001μmol/L group. The level of EGF in 125 μmol group of 72h was obviously raised compared with 24h, but there was no obvious difference compared with 48h. In 250μmol/L group, 72h was obviously raised compared with 24h, but there was no obvious difference compared with 48h. The level of 72h was obviously raised compared with both 24h and 48h in 500μmol/L group. In addition, 125μmol group and 250μmol/L group had no obvious difference, and 500μmol/l, group was obviously raised, all compared with control group at 24h ;the 125μmol/1, group had no obvious difference, but 250μmol/L group and 500μmol/L group were hoth obviously raised compared with control group at 48h. The 125μmol/L, 250μmol/L and 500μmol/L groups were all obviously raised compared with control group at 72h. Conclusion Sodium ferulate which is an active ingredient of Chinese angelica can improve the expressive level of EGF in human dermal fibroblasts, and have prominent dose dependence and time dependence.