酸性成纤维细胞生长因子及其改构体对培养大鼠RGCs存活与生长的影响

目的 观察不同质量浓度aFGF、MaFGF对培养的大鼠视网膜神经节细胞(RGCs)生长的影响,及aFGF的神经保护活性是否依赖促有丝分裂活性.方法 用不同质量浓度的aFGF和MaFGF培养RGCs.免疫细胞化学染色,计数免疫阳性细胞,计算轴突生长细胞百分率,MTT法进行检测.结果 培养第3 d,大部分存活细胞为Thy-1免疫细胞化学反应阳性,5 d、7 d后逐渐减少.实验组存活时间3～4 d;培养第3 d,MTT法见各组A值与对照组比较差异无统计学意义(P＞0.05),培养第5 d、7 d,差异有统计学意义(P＜0.01);培养相同天数不同质量浓度的aFGF及MaFGF比较,差异均有统计学意义(P＜0.05);相同质量浓度的aFGF各组与MaFGF各组比较,差异无统计学意义(P＞0.05).结论 aFGF、MaFGF均能促进培养大鼠RGCs的存活,并延长其存活时间;aFGF保护及促进RGCs存活的作用不依赖其有丝分裂活性.

Effect of acidic fibroblast growth factor and modified acidic fibroblast growth factor on rats' retinal ganglion cells in vitro

Objective To observe the effect of acidic fibroblast growth factor(aFGF)and modified acidic fibroblast growth factor(MaFGF)on the growth and survival of retinal ganglion cells(RGCs)in vitro.MethodsThe retinas of 30 3-day old Sprague-Dawley rats were prepared into suspension with 0.25% trypin digestion.Cultured RGCs were identified with immunohistochemistry using anti-rat Thy-1 monoclonal antibody after cultured 3,5 and 7 days.The numbers of immuntional positive cells of immunostaining and the rate of cells with axon were counted.25,100,400 ng/mL aFGF was added in A1,A2 or A3 group respectively,and the same quantity of MaFGF was used in B1,B2 or B3 group,respectively,and DMEM was added in control group.The A value of living RGCs were tested by methylthio-tetrazole colorimetric microassay in 3,5,7 days of culture.ResultsCultured RGCs showed the brown-yellow staining in cytoplasm and axon of cells by the application of anti-rat Thy-1 monoclonal antibody.The number of RGCs was decreased with prolong of culture time.No significant difference in the number of RGCs and the rate of cells with axon was found among the control,aFGF and MaFGF groups in culture for 3 days(P>0.05),but considerable difference was seen in culture 5 and 7 days groups(P<0.05).The survival time of living RGCs prolonged to 3 to 4 days in aFGF and MaFGF groups compared with the control groups.The A value of living RGCs in culture for 3 days was not obviously different among aFGF,MaFGF and control groups but was significant different in the 5th day and 7th day(P<0.01).In aFGF groups,the A values of A2 group was evidently higher than that of A1 and A2 groups with the significant difference in the same time point(P<0.05),and in MaFGF group,that of B2 group was higher than B1 and B2 groups(P<0.05).The differences among the same concentration of aFGF and MaFGF group at the same culture days were insignificant(P>0.05).ConclusionBoth aFGF and MaFGF could facilitate survival of RGCs in vitro.aFGF and MaFGF have no effect on the conformation and volume of RGCs.