维甲酸调控IGF-2基因对软骨细胞促凋亡的作用  

PRO-APOPTOTIC EFFECT OF RETINOIC ACID ON CHONDROCYTE THROUGH REGULATION ON GENE EX-PRESSION OF IGF-2

作　　者：郭磊[1] 赵玉岩[2] 张世亮[1] 刘魁[1] 高晓宇[1] 

机构地区：[1]中国医科大学附属第一医院骨科,沈阳110001 [2]中国医科大学附属第一医院内分泌科,沈阳110001

出　　处：《中国修复重建外科杂志》2008年第2期217-220,共4页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(30500414);辽宁省教育厅高等学校科学研究资助项目(05L508,20061010)~~

摘　　要：目的探讨维甲酸对软骨细胞凋亡的影响及细胞内IGF-2的表达规律。方法1月龄雄性中国白兔1只,体重约500g。取兔双膝关节股骨髁软骨,采用胰酶消化法体外培养白兔软骨细胞。取第2代软骨细胞,培养液内加入终浓度为1×10-6mol/L的全反式维甲酸(all-trans-retinoic acid,ATRA)25μL(实验组),作用24h;对照组加入25μLDMEM液作为对照。采用细胞免疫组织化学法观察软骨细胞中Ⅱ型胶原的分泌变化,流式细胞仪检测软骨细胞凋亡率,RT-PCR半定量分析软骨细胞中IGF-2mRNA的表达,Western blot印迹杂交鉴定软骨细胞中IGF-2蛋白质表达。结果实验组ATRA抑制了软骨细胞中Ⅱ型胶原的表达。流式细胞仪检测实验组软骨细胞凋亡率为21%±2%,较对照组(5%±1%)增加了5倍;RT-PCR检测实验组IGF-2mRNA的表达为0.2±0.1,较对照组(0.8±0.2)降低75%;Western blot印迹杂交分析发现,实验组软骨细胞IGF-2的表达为0.3±0.1,较对照组(0.7±0.2)降低了57%;各指标组间比较差异均有统计学意义(P<0.05)。结论维甲酸可能通过抑制软骨细胞靶基因IGF-2的表达,负性调节软骨细胞分泌和增殖功能,促进软骨细胞凋亡。Objective To investigate the effect of retinoic acid （RA） on cell apoptosls and gene regulation of IGF-2 in chondrocyte. Methods One 1-month-old Chinese rabbit weighted 500 g was used in this experiment. The chondrocyte from rabbit knee were cultured by enzyme digestion. Twenty-five μL all-trans-retinoic acid （ATRA） （1 ×10^6 mol/L） were added in the media of cultured chondrocyte for 24 hours as experimental group, while 25 μL DMEM were added as control group. The secretion of collagen Ⅱ was observed by immunohistochemistry method, cell apoptosis was detected by flow cytometry, IGF-2 mRNA and protein expression in chondrocyte were detected by RT-PCR and Western blot analysis. Results The expression of collagen Ⅱ was down-regulated by ATRA in the experimental group. The cell apoptosis in chondrocyte exposed to ATRA at 1×10^6 mol/L was 21%±2%, which increased 5 times compared with the control group （5% ±1%）. The IGF-2 mRNA and protein level in the experimental group were decreased 75% and 57%, respectively, compared to the control group. There were significant difference between the experimental group and control group in each index （P 〈 0.05）. Conclusion RA may down-regulate the secretion and cell proliferation, but up-regulate the cell apoptosis in chondrocyte. The apoptotic effect may carry out through inhibiting the IGF-2 expression of chondrocyte.

关 键 词：维甲酸 软骨细胞 凋亡 IGF-2 基因表达 

分 类 号：R684[医药卫生—骨科学]