PDGF-BB和CTGF在体外对人皮肤成纤维细胞增殖的影响

目的探讨重组人血小板衍生生长因子（rhPDGF-BB）和重组人结缔组织生长因子（rhCTGF）单独或联合应用对体外培养的人皮肤成纤维细胞（human skin fibroblast,HSF）增殖的影响,为寻求快速扩增HSF的方法提供参考依据。方法取手术中切除的正常皮肤,以常规组织块法体外获得皮肤成纤维细胞。取生长良好的第4~7代细胞种入96孔板,分实验组和对照组,分别加入不同浓度的rhPDGF-BB、rhCTGF或rhPDGF-BB＋rhCTGF作用。于第1、3、6、9d采用四唑盐（MTT）法分别测定各组吸光度A值,观察上述两种生长因子对细胞的增殖作用。结果 rhPDGF-BB能明显促进体外培养的HSF增殖,在1~9d,0.1ng/ml~100ng/ml范围内呈浓度时间依赖关系。rhCTGF也能明显促进HSF增殖,除第1d最大效应浓度为50ng/ml,其余时间点在10ng/ml~100ng/ml范围内呈浓度时间依赖关系。而rhCTGF150ng/ml组的增殖效果相对100ng/ml组略成下降趋势。rhPDGF-BB与rhCTGF最大与最小效应联用组亦成浓度时间依赖关系。最大效应联用组的增殖效果与以上两种生长因子单独应用比较均具有统计学意义（P〈0.05）。最小效应联用组与rhCTGF10ng/ml组比较有统计学意义（P〈0.05）,而与rhPDGF-BB0.1ng/ml组比较无统计学意义（P〉0.05）。另外,最大效应浓度的两生长因子单独应用时,rhPDGF-BB促增殖效果优于rhCTGF（P〈0.05）。结论 rh-PDGF-BB和rhCTGF均可促进体外培养的HSF增殖,在一定范围内呈浓度时间依赖关系,而且前者优于后者。两者联合应用对HSF增殖具有协同效应。

The influence of PDGF-BB and CTGF on the proliferation of human skin fibroblast in vitro

Objective To evaluate the influence of combination of recombinant human Plateled Derived Growth Factor-BB（rhPDGF-BB） and combination human Connective Tissue Growth Factor（rhCTGF） and individuality application on the proliferation of Human Skin Fibroblast（HSF）,and to supply a method of rapid proliferation of HSF in vitro.Methods The biopsies came from the healthy donors accepted cosmetic surgery,and the HSF were obtained by tissue culture.rhPDGF-BB,rhCTGF,or rhPDGF-BB＋rhCT-GF of different concentrations were added into the HSF which were passed four-seven genetation,and culture.The dose-response re-lationship was studied at 1d,3d,6d and 9d by the MTT colorimetric method.Results Compared with control group,rhPDGF-BB in-duced proliferation of HSF in a concentration-and time dependent manner with the concentration from 0.1ng/ml to 100ng/ml in 9 days.When compared with control group,rhCTGF can also stimulated the cell proliferation significantly.And within 9 days,the func-tion of rhCTGF was maximum at 100ng/ml of the 9th day,minimum at 10ng/ml of the 1st day,and the proliferation activity of the concentration of 100ng/ml was better than 150ng/ml.But in the first day,the best concentration was 50ng/ml.The functions of com-bined with the two growth factors were also concentration-time depended,and better than use it alone（P〈0.05）,except the group of rhPDGF-BB of 0.1ng/ml（P〉0.05）.By the way,the function of rhPDGF-BB was better than rhCTGF at 100ng/ml（P〈0.05）.Conclusion Both of rhPDGF-BB and rhCTGF can promote the proliferation of HSF in vitro,and the function can be concentration-time depend-ed,and the former is better.The combination of two growth factors has a synergistic action on the promotion of HSF.