人重组骨形态发生蛋白-2和碱性成纤维细胞生长因子在诱导成骨中的调节作用及其机制  

Regulatory function of recombinant human bone morphogenetic protein 2 and basic fibroblast growth factor in the induced osteogenesis and their regulatory mechanisms

作　　者：杜俊杰[1] 罗卓荆[1] 胡蕴玉[1] 李明全[1] 袁志[1] 曹艳杰[2] 崔庚[1] 李丹[1] 吕荣[1] 王建波[3] 

机构地区：[1]解放军第四军医大学西京医院全军骨科研究所,陕西省西安市710032 [2]解放军第四军医大学西京医院心血管内科实验室,陕西省西安市710032 [3]解放军第四军医大学西京医院病理科,陕西省西安市710032

出　　处：《中国临床康复》2004年第8期1448-1449,共2页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助项目(39600148)~~

摘　　要：目的:采用原位杂交的方法检测I、II型胶原mRNA的变化及其变化规律,对骨组织中的碱性磷酸酶(ALP)活性进行测定,以分析人重组骨形态发生蛋白-2(rhBMP-2)和碱性成纤维细胞生长因子(bFGF)在诱导成骨中的调节作用及其调节机制。方法:BALB/c小鼠110只采用掷硬币方法随机分为2组,每组55只,设立rhBMP-2/bFGF为实验组,rhBMP-2为对照组,分别于术后3～21d共8个时间点取材,采用原位杂交方法对新生骨组织中的I,II型胶原mRNA进行检测;对ALP活性进行定量分析,观察2组在诱导成骨中I,II型胶原mRNA及ALP的表达情况。结果:①II型胶原mRNA的出现是和成软骨、软骨细胞的出现相伴随的。②作为成骨细胞成熟标志物的I型胶原mRNA、ALP在软骨形成期即有表达,但是随着成骨细胞的出现,骨组织的形成I型胶原mRNA仍表现为高表达,而ALP的表达则呈下降趋势。③实验组I,II型胶原mRNA及ALP的表达早于对照组,在术后7,14,21d,实验组ALP分别为(63.85±6.87),(27.12±4.23),(8.93±1.49)IU/g;对照组ALP分别为(27.26±4.13),(70.65±5.92),(39.46±4.72)IU/g(t=12.40,20.85,17.52,P<0.01)。结论:bFGF增强了rhBMP-2诱导成骨中I、II型胶原mRNA及ALP的表达。AIM:To detect the changes and changing rules of type I, II collagen mRNA expression with in situ hybridization,and to assay the activity of alkaline phosphatase(ALP) in the bone tissue so as to analyze the regulatory function of bone morphogenetic protein 2(rhBMP 2) and basic fibroblast growth factor(bFGF) during osteogenesis and their regulatory mechanisms. METHODS:Totally 110 BALB/c mice were randomly divided into two groups by throwing coins(55 in each group).Mice receiving rhBMP 2/bFGF served as the trial group and those receiving rhBMP 2 served as the control group.In order to detect the expression of collagen I, II mRNA and ALP in osteogenesis induced by rhBMP 2/ bFGF and rhBMP 2,in situ hybridization analysis was performed at 3, 5, 7, 9, 11, 14, 17 and 21 days after the implantation.Meanwhile,ALP activity was analyzed quantitatively,and the expression of collagen I, II mRNA and ALP was observed. RESULTS:①The formation of type II collagen mRNA was accompanied by that of chondroblasts and chondrocytes.②Type I collagen mRNA and ALP, markers of osteoblast differentiation,were expressed at the early stage of chondrogenesis,but type I collagen mRNA was still expressed at higher level,and ALP decreased gradually with the formation of osteoblasts and osteogenesis.③Expression of collagen I, II mRNA and ALP in the trial group was higher than that in the control group,ALP was (63.85±6.87),(27.12±4.23) and (8.93±1.49) IU/g in the trial group and (27.26±4.13),(70.65±5.92),(39.46±4.72) IU/g in the control group(t=12.40,20.85,17.52,P< 0.01) respectively at 14, 17 and 21 days after the implantation. CONCLUSION:bFGF enhances the expression of collagen I,II mRNA and ALP in osteogenesis induced by rhBMP 2.

关 键 词：人重组骨形态发生蛋白-2 碱性成纤维细胞生长因子 成骨诱导 调节作用 原位杂交 胶原 碱性磷酸酶活性 

分 类 号：R318[医药卫生—生物医学工程]