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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 investigate the effect of histatin 1 (Hst1) on the proliferation and migration of human fibroblasts. Methods Human fibroblasts were subcultured by conventional methods and divided into blank control group (DMEM + 1% bovine serum), group A (100μg / ml Hst1), group B (30μg / ml Hst1), group C mlHst1). The effect of different concentrations of Hst1 (3 ~ 100μg / ml) on the proliferation of human fibroblasts at different time points (24,48,72 h) was observed by cell counting method. The fused human fibroblasts were divided into Mitomycin C treatment group and mitomycin C untreated group were divided into blank control group (DMEM + 1% bovine serum), group Ⅰ (30μg / ml Hst1), group Ⅱ (10ng / ml rhEGF), group Ⅲ (30μg / ml Hst1 + 10 ng / ml rhEGF), group Ⅳ (15μg / ml Hst1 + 5 ng / ml rhEGF) Effect of Hst1 on the migration of human fibroblasts in vitro. Results ① Hst1 at different concentrations promoted the proliferation of human fibroblasts with no significant dependence on concentration and time. The number of cells in groups A and B at 24 h was significantly higher than that in other groups (P <0.01) Promote healing of cell scratches wound healing rate of about 37.7% 8 h scratches, when combined with rhEGF promote wound healing rate was higher than the single drug; use of mitomycin C to exclude cell proliferation, 30μg / ml Hst1 more blank The control group did not promote cell migration (P> 0.05), the healing rate of scratches was lower than that of mitomycin C untreated group, but there was no significant difference between the two groups Statistical difference (P> 0.05). Conclusion Hst1 can promote the proliferation of human fibroblasts, but the migration function is not obvious. The use of Hst1 in combination with rhEGF has a synergistic effect on wound healing of human fibroblasts.