富组蛋白1促皮肤创伤愈合的细胞学研究

Cytological Study on Histatin 1 in Healing Promotion of Skin Wound

目的了解富组蛋白1(histatin1,Hst1)对人表皮细胞(human adult skin keratinocytos,HaCaT)、人成纤维细胞株增殖和迁移功能的影响。方法细胞增殖实验:(1)将HaCaT细胞、人成纤维细胞均分为空白对照组(1640/DMEM+1%新生牛血清)、Ⅰ组(100μg/ml Hst1)、Ⅱ组(30μg/ml Hst1)、Ⅲ组(3μg/ml Hst1),比较两种细胞的增殖数量;(2)细胞划痕实验:将两种细胞分为空白对照组(1640+1%新生牛血清)、A组(30μg/ml Hst1)、B组[10ng/ml人重组表皮生长因子(recombinant human epidermalgrowth factor,rhEGF)]、C组(30μg/ml Hst1+10ng/ml rhEGF)、D组(15μg/ml Hst1+5ng/ml rhEGF)、E组(15μg/ml Hst1+10ng/ml rhEGF),比较两种细胞体外创面愈合速度。结果 (1)3～100μg/ml的Hst1能够促进HaCaT增殖,72h时Ⅰ组细胞数高于空白对照组、Ⅱ组及Ⅲ组(P<0.01);3～100μg/ml的Hst1能够促进人成纤维细胞增殖,24h时Ⅰ、Ⅱ组细胞数高于其余各组(P<0.01);(2)30μg/ml Hst1能促进HaCaT细胞迁移,16h时A组划痕愈合率高于空白对照组(P<0.01),C、D组高于A组(P<0.05);30μg/ml Hst1能促进人成纤维细胞划痕创面愈合,与rhEGF联合应用时划痕愈合率高于单独用药。结论 Hst1能够促进HaCaT细胞和人成纤维细胞增殖和迁移,与rhEGF联合应用时对其促进体外创伤愈合具有协同作用。

Objective To observe the effect of histatin1（Hst1）and recombinant human epidermal growth factor（rhEGF） on the proliferation and migration of human epidermal cells（HaCaT） and human skin fibroblasts.Methods Cells prdiferation assay：（1） Cells were randomly divided into control group（1640＋1%NBS）,100 μg/ml Hst1 group（group Ⅰ）,30 μg/ml Hst1 group（group Ⅱ）,3 μg/ml Hst1 group（group Ⅲ）.The ability of cell proliferation was observed;（2） Cell scratch experiments：cells was randomly divided into control group（1640/DMEM ＋1%NBS）,30 μg/mlHst1 group（group A）,10ng/mlrhEGF group（group B）,30 μg/ml Hst1 ＋ 10 ng/ml rhEGF group（group C）,15 μg/ml Hst1 ＋ 5 ng/ml rhEGF group（group D）,15 μg/ml Hst1 ＋10 ng/ml rhEGF group（group E）.The effects of Hst1 on healing area rates of cells were compared between the two groups.Results（1） The ability of cell proliferation was significantly enhanced by 3～100 μg/ml Hst1 groups.The cell count of HaCaT in group Ⅰ at 72 h was significantly higher than that in the control group,group Ⅱ and group Ⅲ（P0.01）.The ability of human skin fibroblasts proliferation was significantly enhanced by group Ⅱ and group Ⅰ at 24 h（P0.01）.（2） The healing area rate of HaCaT cells increased remarkable by 30μg/mlHst1,and the healed area rate at 16h was significant more than control group（P0.01）.The healing area rate in combination groups（C,D）was significantly more than that in group A（P0.05）.The healing area rate of human skin fibroblasts increased remarkable by 30 μg/ml Hst1.A joint application of Hst1 and rhEGF has a synergistic effect in promoting the external wound healing.Conclusion Hst1 can induce the proliferation and migration of human epidermal cells and human skin fibroblasts,and a joint application of rhEGF and with Hst1 has a synergistic effect in promoting the external wound healing.