成纤维细胞生长因子2不同作用时长对软骨细胞的影响  

Effects of FGF2 administration for different time periods on chondrocytes

作　　者：曹一汀[1] 杨文裕 张哲[1] 史艳萍[2] 李学敏[1] 张其清[1] 

机构地区：[1]中国医学科学院北京协和医学院生物医学工程研究所,天津市生物医学材料重点实验室,天津300192 [2]天津理工大学化学化工学院,300384

出　　处：《国际生物医学工程杂志》2017年第2期91-97,I0005,共8页International Journal of Biomedical Engineering

基　　金：天津市应用基础与前沿技术研究计划（重点项目）（14JCZDJC38200）

摘　　要：目的 比较成纤维细胞生长因子2（FGF2）体外不同作用时长对兔关节软骨细胞增殖及基因表达变化的影响.方法 分离、培养兔膝关节软骨细胞.取第3代软骨细胞分为3组进行研究：短时间（1 d）作用组细胞经FGF2处理1d后转移至无FGF2的培养基中继续培养7d;长时间（7 d）作用组细胞用含FGF2的培养基连续培养7d;对照组细胞用无FGF2的培养基连续培养7d.分别于培养过程中的第1、3、7天,收集软骨细胞进行细胞增殖情况检测;提取软骨细胞mRNA,并通过实时荧光定量PCR检测骨形态发生蛋白2（BMP2）、骨形态发生蛋白4（BMP4）、Ⅱ型胶原α1（COL2A1）和性别决定区Y框蛋白9（SOX9）的表达情况;采用免疫荧光染色测定软骨细胞Ⅱ型胶原的含量.结果 与对照组相比,不同时长的FGF2处理均能促进软骨细胞的增殖,且短时间作用组和长时间作用组间差异无统计学意义（P＞0.05）.实时荧光定量结果显示,不同的处理方式引起了不同的基因表达变化,在连续培养7d后,与对照组和长时间作用组比较,短时间作用组的BMP2、BMP4、COL2A1和SOX9基因表达均上调,差异均具有统计学意义（均P＜0.05）.免疫荧光染色结果表明,连续培养7d后,短时间作用组的Ⅱ型胶原含量明显高于对照组和长时间作用组.结论 FGF2短时间作用于软骨细胞较长时间作用更有利于软骨细胞表型的维持和细胞外基质的合成.Objective To study the effects of fibroblast growth factor 2 （FGF2） on the proliferation and gene expression profiles of rabbit articular chondrocytes in vitro after different time periods of stimulation.Methods The chondrocytes were isolated and cultured in vitro,and the 3rd generation cells were harvested.Cells were divided into three groups.In the group 1 （FGF2 short-time action group）,chondrocytes were cultured in medium with FGF2 for one day,and then transferred to fresh culture medium without FGF2 and cultured for another 6 days.In the group 2 （FGF2 long-time action group）,chondrocytes were cultured in medium with FGF2 for 7 days.In the Group 3 （control group）,chondrocytes were cultured in culture medium without FGF2 for 7 days.After culture for 1,3,and 7 days,the proliferation of chondrocytes in the all groups was detected respectively.Following extraction of mRNA,the gene expressions of BMP2,BMP4,SOX9 and COL2A1 of the chondrocytes in the all groups were determined by quantitative real-time polymerase chain reaction （qRT-PCR）.The content of type Ⅱ collagen was measured via immunofluorescence staining.Results Compared with the control group,FGF2 promoted the proliferation of chondrocytes in the short-and long-time action groups and there was no significant difference between the two FGF2-treated groups.The results of qRT-PCR indicated that different treatment induced different gene expression profile.Particularly,compared with the control group and the FGF2 long-time action group,the expression of BMP2,BMP4,SOX9 and COL2A1 in the short-time action group were significantly upregulated at the 7th day.Immunofluorescence intensity of type Ⅱ collagen in the group 1 was stronger than that in the control group and group 2.Conclusions Different administration of FGF2 for different time periods induced different responses of chondrocytes.Short-term FGF2 stimulation was more beneficial to maintain the phenotype of chondrocytes and the synthesis of extracellular matrix.

关 键 词：成纤维细胞生长因子2 软骨细胞 长时间作用 短时间作用 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]