CAJ | 학술논문

目的探讨骨骼发育畸形的大鼠软骨组织内胰岛素样生长因子(insulin-like growth factors,IGFs)家族成员IGF-2的表达规律.方法应用环境类致畸因子二恶英中毒性最强的四氯二苯并二恶英,即2,3,7,8-四氯一二苯并-对-二恶英(2,3,7,8-tetrachlorodibenzo-p-dioxin,TCDD)构建先天性大鼠骨骼发育畸形动物模型.应用光镜和透射电镜观测骨骼畸形大鼠的足部软骨组织病理学改变.免疫组织化学染色分析大鼠软骨组织IGF-2蛋白的表达.应用TCDD干预体外培养的大鼠软骨细胞,采用RT-PCR及Western印迹杂交检测软骨细胞内的IGF-2 mRNA及蛋白质的表达水平.结果 TCDD(15 μg/kg)诱导实验组33.3%的胎鼠出现单一或多种畸形,包括:内翻足、小脑畸形、腭裂、无尾畸形等,在骨骼畸形胎鼠的足部软骨发生带缩小,软骨细胞数量减少,软骨细胞核内粗面内质网扩张,核基质降解,线粒体嵴紊乱.15 μg/kg剂量的TCDD使畸形胎鼠软骨细胞内IGF-2蛋白质的表达明显降低.TCDD(1 × 10-8 mol/L)作用体外培养的大鼠软骨细胞24 h,细胞内IGF-2的基因转录和翻译水平分别减少80%和60%,P＜0.05.结论 IGF 2在大鼠软骨细胞内的低表达可能与TCDD对骨骼发育的致畸作用密切相关.
목적 탐토골격발육기형적대서연골조직내이도소양생장인자(insulin-like growth factors,IGFs)가족성원IGF-2적표체규률.방법 응용배경류치기인자이악영중독성최강적사록이분병이악영,즉2,3,7,8-사록일이분병-대-이악영(2,3,7,8-tetrachlorodibenzo-p-dioxin,TCDD)구건선천성대서골격발육기형동물모형.응용광경화투사전경관측골격기형대서적족부연골조직병이학개변.면역조직화학염색분석대서연골조직IGF-2단백적표체.응용TCDD간예체외배양적대서연골세포,채용RT-PCR급Western인적잡교검측연골세포내적IGF-2 mRNA급단백질적표체수평.결과 TCDD(15 μg/kg)유도실험조33.3%적태서출현단일혹다충기형,포괄:내번족、소뇌기형、악렬、무미기형등,재골격기형태서적족부연골발생대축소,연골세포수량감소,연골세포핵내조면내질망확장,핵기질강해,선립체척문란.15 μg/kg제량적TCDD사기형태서연골세포내IGF-2단백질적표체명현강저.TCDD(1 × 10-8 mol/L)작용체외배양적대서연골세포24 h,세포내IGF-2적기인전록화번역수평분별감소80%화60%,P＜0.05.결론 IGF 2재대서연골세포내적저표체가능여TCDD대골격발육적치기작용밀절상관.
Objective To investigate the gene expression of insulin-like growth factors-2(IGF2)in fetal rat with congenital skeleton malformation.Methods The fetal rat models with congenital skeleton malformation were induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD)to the pregnant Wistar rats(day 10).Histopathologic characteristics of cartilaginous tissue in fetal foot were detected with light microscope and transmission electron microscope.IGF-2 protein level in cartilaginous tissue was analyzed by immunocytochemical methods and image analysis.The IGF-2 mRNA and protein level in TCDD-treated chondrocytes in vitro were detected by reverse transcription polymerase chain reaction and western blotting analysis.respectively.Results TCDD(15 μg/kg)induced single or coalescent developmental skeleton defect in fetal rat(33.3%),such as cross-foot,cleft spine,cleft palate,rumplesand so on,The chondrogenesis center of tarsal bones and quantity of chondrocyte were decreased.There were expanding rough endoplasmic reticulum,degradation of nuclear matrix,and disorder of mitochondria cristae in cell nucleus of chondrocyte.IGF-2 protein level in TCDD-treated cartilaginous'tissue was decreased significantly.In chondrocyte treated with TCDD(1 × 10-8 mol/L)in vitro,the expression of IGF-2 mRNA and protein were decreased 80%and 60%,respectively(P＜0.05).Conelusions The low expression of IGF-2 in chondrocyte may be related to the TCDD-induced developmentalskeleton defect.