血小板源性生长因子BB对体外培养兔角膜成纤维细胞增殖和细胞周期的影响

Effects of recombinant human PDGF-BB on the proliferation and cell cycle of cultured rabbit corneal fibroblasts

目的探讨重组人血小板源性生长因子BB(recombinant human platelet-derived growth factor-BB,rhPDGF-BB)对体外培养的兔角膜成纤维细胞增殖和细胞周期的影响。方法体外培养兔角膜成纤维细胞,分实验组和对照组,对照组弃原培养液,加无血清培养液继续培养,实验组用不同浓度的rhPDGF-BB1μg·L-1、10μg·L-1、100μg·L-1、1000μg·L-1分别作用于兔角膜成纤维细胞24h、48h、72h,用MTT比色法检测rhPDGFBB对兔角膜成纤维细胞生长的影响,流式细胞仪检测rhPDGFBB对兔角膜成纤维细胞细胞周期的影响。结果rhPDGFBB在1～100μg·L-1浓度范围内与兔角膜成纤维细胞增殖呈剂量效应关系,当浓度增大到1000μg·L-1,促增殖作用减弱。流式细胞术对细胞周期时项分析结果显示,经rhPDGFBB作用后,兔角膜成纤维细胞的DNA合成量(S%)显著升高,G1%降低,反映细胞增殖活力的增殖指数PI值增高。结论rhPDGFBB能促进兔角膜成纤维细胞的增殖和DNA合成,这一作用可能是通过促使处于G1期的兔角膜成纤维细胞进入S期来实现的。

Objective To evaluate the effects of recombinant human platelet-derived growth factor on the proliferation and cell cycle of cultured rabbit corneal fibroblasts（RCF） Methods It was divided into experimental group and control group in cultured rabbit corneal fibroblasts.The control group, deserted DMEM and added DMEM without containing fetal bovine serum, was continued researching. The cultured ceils of experimental group were treated with rhPDGF-BB at varing concentrations of 1μg·L^-1, 10μg·L^-1, 100μg·L^-1,1000μg·L^-1,for 24 hours,48 hours,72 hours respectively.MTT was used to detect proliferating rates of cultured rabbit corneal fibroblasts. Flow cytometry was used to analyze cell cycle. Results The proliferating effects of rhPDGF-BB increased with the concentrations between 1 μg·L^-1 and 100 μg·L^-1 and while with increasing dose, the proliferation effect decreased. Employing the flow cytometry technique we observed the increased S%, the decreased G1 % and the enhanced （ S ＋ G2M） %, when the cells were stimulated by rhPDGF-BB. Conclusion rhPDGF-BB can enhance the proliferation and DNA synthesis of cultured RCF, which may be accomplished by accelerating Gl to S phase transition. [Rec Adv Ophthalmol 2005; 25 （5） ： 389-392 ]