富组蛋白1促进人成纤维细胞增殖和迁移

Histatin1 improves proliferation and migration in human human skin fibroblasts

目的了解富组蛋白1(histatin1,Hst1)对人成纤维细胞增殖和迁移功能的影响。方法将人成纤维细胞按常规方法传代培养,分为空白对照组(DMEM+1%小牛血清)、A组(100μg/ml Hst1)、B组(30μg/ml Hst1)、C组(3μg/mlHst1),细胞计数法观察不同浓度Hst1(3～100μg/ml)在不同时相点(24、48、72 h)对人成纤维细胞增殖功能的影响;将已融合的人成纤维细胞分为丝裂霉素C处理组及丝裂霉素C未处理组,各组中再次分为空白对照组(DMEM+1%小牛血清)、Ⅰ组(30μg/ml Hst1)、Ⅱ组(10 ng/ml rhEGF)、Ⅲ组(30μg/ml Hst1+10 ng/ml rhEGF)、Ⅳ组(15μg/ml Hst1+5 ng/mlrhEGF)、Ⅴ组(15μg/ml Hst1+10 ng/ml rhEGF),通过体外细胞划痕实验考察Hst1对人成纤维细胞迁移功能的影响。结果①不同浓度的Hst1能够促进人成纤维细胞增殖,与浓度及时间无明显依赖关系,24 h时间点A、B组细胞数显著高于其余各组(P<0.01);②30μg/ml Hst1能促进细胞划痕创面愈合,8 h划痕愈合率约37.7%,与rhEGF联合应用时促划痕愈合率高于单独用药;利用丝裂霉素C排除细胞增殖作用后,30μg/ml Hst1较空白对照组无促进细胞迁移功能(P>0.05),划痕愈合率较丝裂霉素C未处理对应组下降,但无统计学意义,且与rhEGF联合应用时促划痕愈合率与单独用药无统计学差异(P>0.05)。结论 Hst1能够促进人成纤维细胞增殖,但促迁移功能不明显。Hst1联合rhEGF使用时对人成纤维细胞划痕创伤愈合具有协同促进作用。

Objective To determine the effect of histatin1（Hst1） and recombinant human epidermal growth factor（rhEGF） on the proliferation and migration in human skin fibroblasts.Methods Human skin fibroblasts was subcultured,and then randomly divided into control group（DMEM＋1%NBS）,A group（100 μg/ml Hst1）,B group（30 μg/ml Hst1） and C group（3 μg/ml Hst1）.Cell proliferation was observed by growth curve at 24,48 and 72 h after corresponding treatment.Human skin fibroblasts was also randomly divided into control group（DMEM＋1%NBS）,Ⅰgroup（30 μg/ml Hst1）,Ⅱgroup（10 ng/ml rhEGF）,Ⅲgroup（30 μg/ml Hst1＋10 ng/ml rhEGF）,Ⅳgroup（15 μg/ml Hst1＋5 ng/ml rhEGF）,and Ⅴgroup（15 μg/ml Hst1＋10 ng/ml rhEGF）.The ability of cell migration was evaluated by cell scrath test.Results ① The cell proliferation was significantly enhanced in B group and A group at 24 h（P0.01）.②Hst1 at 30 μg/ml resulted in remarkable increased cells in the healing area,with a healing rate of 37.7% at 8 h.The healing area rate in combination groups（Hst1＋rhEGF） was significantly higher than that in 10 ng/ml rhEGF group or 30 μg/ml Hst1 group.After Mitomycin C treatment,which restrained the cells proliferation,the cells healing area was more elevated in 10 ng/ml rhEGF group,15 μg/ml Hst1 ＋ 5 ng/ml rhEGF group and 15 μg/ml Hst1 ＋10 ng/ml rhEGF group,but not in 30 μg/ml Hst1 group when compared with that in control group.The healing area was declined at 8 h than that of Mitomycin C untreated,but the difference was no significant.The healing area in combination groups（Hst1＋rhEGF） had no significantly difference with that in 10 ng/ml rhEGF group or 30 μg/ml Hst1 group.Conclusion Hst1 can induce the proliferation of human skin fibroblasts,but exert not significant effect on migration.rhEGF combined with Hst1 shows a synergic effect on wound healing.