血小板源性生长因子和bFGF促进猫角膜内皮细胞增生的协同作用研究

Synergistic effect of platelet-derived growth factor BB and basic fibroblast growth factor on cat corneal endothelial cell proliferation

目的观察血小板源性生长因子(PDGF-BB)和碱性成纤维细胞生长因子(bFGF)对猫角膜内皮细胞(CECs)增生的影响及其联合应用对细胞增生是否有协同作用。方法猫CECs原代培养,传1代后,分别在培养液中加不同浓度的PDGF-BB和bFGF,加药后第1、3、5d分别利用MTT法测定在490nm处的吸光值来判断CECs的增生情况;同时应用倒置相差显微镜观察细胞形态,扫描电镜检测细胞超微结构。结果与对照组比较,加药后第1、3、5d,PDGF-BB和bFGF均能增加培养的猫CECs的数量,最有效质量浓度分别为100ng/ml和10ng/ml,二者联合培养组猫CECs的增生能力更强。结论PDGF-BB和bFGF可促进猫CECs的增生,二者具有协同作用。

Objective Platelet derived growth factor BB （PDGF-BB） has the function of promoting cell differentiation. Basic fibroblast growth factor （bFGF） could not only promote cell differentiation but also stimulate cell chemotaxis and migration. This experiment investigated the function of PDGF-BB and bFGF as well as their cooperative effects on primary cultured cat corneal endothelial cells（CECs） proliferation. Methods Cultured cat CECs were treated with PDGF-BB （ 1, 10,and 100 ng/ ml） and bFGF （ 1,10 ,and 100 ng/ml） separately or both at 1,3,5 days after primary culture. Cat CECs were cultured in DMEM medium containing O. 5% FCS in control group. Proliferation of cultured cat CECs was assayed by modified tertrozalium salt （MTT） method. The morphologic changes of cultured cells were observed with inverted phase contrast microscope and scanning electron microscope. Results In comparison with control group, the numbers of cat CECs was significantly increased with increasing concentration of PDGF-BB and duration of treatment and a similar reaction was found in bFGF treading group. The proliferating peak of CECs was reached in 100 ng/ml PDGF-BB or 10 ng/ml bFGF group,and the effects of combined PDGF-BB and bFGF on the proliferation of CECs were more distinct than PDGF-BB and bFGF alone. The CECs treated with both PDGF-BB and bFGF became confluent,and cell branches were invisible in comparison with those treated with PDGF-BB and bFGF alone on the 5th day. Conclusion PDGF-BB and bFGF could promote proliferation of cat CECs at a dose- and time-dependent manner, and these two factors have a synergistic effects in enhancing proliferation of cells.