人Ad-BMP-2基因转染兔骨髓基质干细胞对其Ⅱ型胶原和碱性磷酸酶表达的影响

doi:吉林省科技厅资助课题

人Ad-BMP-2基因转染兔骨髓基质干细胞对其Ⅱ型胶原和碱性磷酸酶表达的影响

韩冬^1△，李建军²，张巨¹，孙鸿斌¹，顾家祥¹，王健3，刘建国²，王国君^1*

1.吉林大学中日联谊医院手外科，吉林长春130031；2.吉林大学第一医院骨科，吉林长春130021；3.温州医学院附属一院骨科，浙江温州325000

收稿日期:2004-01-13 修回日期:1900-01-01 出版日期:2005-01-28 发布日期:2005-01-28
通讯作者: 王国君

Effects of BMP-2 gene transduced rabbit bone marrow stromalcells on expressions of collagen type Ⅱ and AKP

HAN Dong^1△, LI Jian-jun², ZHANG Ju¹, SUN Hong-bin¹,GU Jia-xiang¹, WANG Jian3, LIU Jian-guo²,WANG Guo-jun^1*

1. Department of Hand Surgery, China-Japan Union Hospital, Jilin University, Changchun 130031, China;2. Department of Orthopedics, First Hospital, Jilin University, Changchun 130021, China;3. Department of Orthopedics, First Hospital ,Wenzhou Medical College, Wenzhou 325000, China

Received:2004-01-13 Revised:1900-01-01 Online:2005-01-28 Published:2005-01-28
Contact: WANG Guo-jun

摘要/Abstract

摘要： 目的：用人骨形态发生蛋白2腺病毒表达载体（Ad-BMP-2）转染兔骨髓基质干细胞（BMSC），观察Ad-BMP-2转染对细胞分化的影响。方法：将Ad-BMP-2转染体外培养的兔BMSC，7 d后利用原位杂交和免疫组化方法检测细胞内BMP-2及Ⅱ型胶原的表达，同时行碱性磷酸酶染色。结果：BMP-2和Ⅱ型胶原原位杂交法显示实验组细胞胞浆中有棕黄色阳性杂交信号，对照组阴性。BMP-2和Ⅱ型胶原免疫组化法显示实验组细胞胞浆棕黄色阳性着色，胞核阴性表达，对照组阴性；碱性磷酸酶染色见实验组细胞胞浆呈棕黑色阳性着色，可见细小的棕黑色颗粒，对照组染色阴性。结论：Ad-BMP-2可高效转染BMSC，且促进其向成骨细胞和软骨细胞方向转化。

关键词: 人, 骨形态发生蛋白质类, 转染, 干细胞, 胶原Ⅱ型, 碱性磷酸酶

Abstract: Objective Using a recombinant adenoviral vector carrying the human BMP-2 gene (Ad-BMP-2) to transfect rabbit bone marrow stromal cells (BMSC) to study the effects of Ad-BMP-2 transfection on cell differentiation. Methods The rabbit BMSC cultured in vitro were transfected by Ad-BMP-2. Seven days later, the expressions of BMP-2 and collagen type Ⅱ in these cells were determined by in situ hybridization and immunohistochemical analysis, and the alkaline phosphatase (AKP) activity of the BMSC was observed. Results There were positive results in cytoplasm in experimental group: the brown-yellow hybridization signals were observed and that indicated the expressions of BMP-2 and collagen type Ⅱ of Ad-BMP-2 infected BMSC by in situ hybridization, the yellow pigmenting were observed by immunohistochemical analysis but not in nuclei, the brown-black particles were seen by AKP staining. But in control group, these were negative. Conclusion Ad-BMP-2 could infect BMSC with high efficiency and promote the conversion to osteoblast and chondrocyte.

Key words: human, bone morphogenetic proteins, transfection, stem cells, collagen type Ⅱ, alkaline phosphatase

中图分类号:

韩冬，李建军，张巨，孙鸿斌，顾家祥，王健3，刘建国，王国君. 人Ad-BMP-2基因转染兔骨髓基质干细胞对其Ⅱ型胶原和碱性磷酸酶表达的影响[J]. J4, 2005, 31(1): 45-47.

HAN Dong, LI Jian-jun, ZHANG Ju, SUN Hong-bin,GU Jia-xiang, WANG Jian3, LIU Jian-guo,WANG Guo-jun. Effects of BMP-2 gene transduced rabbit bone marrow stromalcells on expressions of collagen type Ⅱ and AKP[J]. J4, 2005, 31(1): 45-47.

[1]	徐丽娜,何小双,辛雯艳,邬超. 骨髓间充质干细胞来源的外泌体对哮喘模型小鼠气道阻力和肺组织病变的改善作用及其机制[J]. 吉林大学学报(医学版), 2026, 52(3): 602-611.
[2]	王玲玉,李瑞丽,张宇,郭美彤,梁亚龙,韩旭,黄晓巍,张扬,于秀华. 人参健智片对认知障碍模型大鼠学习记忆能力的改善作用和海马神经元的保护作用及其机制[J]. 吉林大学学报(医学版), 2026, 52(3): 719-729.
[3]	贾贺强,潘路,王振江,逯家宇,王丹,刘立峰,董营,于春艳. 人参三醇皂苷对D-半乳糖诱导小鼠骨骼肌衰老的改善作用及其机制[J]. 吉林大学学报(医学版), 2026, 52(3): 730-737.
[4]	张佳翔,曹阳,桓晨. HIV-1 Tat蛋白的翻译后修饰对HIV-1转录和复制调控机制影响的研究进展[J]. 吉林大学学报(医学版), 2026, 52(3): 854-862.
[5]	杨如意,王煜灵,杨巍. 人参果胶醇沉组分对小鼠骨髓源性树突状细胞表型及功能的影响[J]. 吉林大学学报(医学版), 2026, 52(2): 375-383.
[6]	翟丽,陈孟,罗建波,张爱利,王良晓,魏颖,张曦. miR-214-3p与EZH2的靶向关系及其对卵巢癌SKOV3细胞增殖、侵袭和凋亡的影响[J]. 吉林大学学报(医学版), 2026, 52(2): 460-468.
[7]	张育添,吴海灵,廖科棋,梁淑铃,李胜男,李友. TAGLN2 shRNA慢病毒载体的构建及其稳定转染细胞系的建立[J]. 吉林大学学报(医学版), 2026, 52(2): 536-542.
[8]	杨慧颖,梁伟虹,徐新茹,孙浩丹,焦昕,王海萍. 骨髓间充质干细胞对心肌样细胞分化诱导、心肌细胞保护及其心肌相关疾病作用的研究进展[J]. 吉林大学学报(医学版), 2026, 52(1): 281-289.
[9]	刘景,王艳,黄旭. 过表达circRNAs修饰牙髓干细胞来源的外泌体对人脐静脉内皮细胞血管生成的促进作用[J]. 吉林大学学报(医学版), 2025, 51(6): 1561-1570.
[10]	苏秋源,赵玲,谭佳佳,莫世恩,周海琴,卢芳芳,韦依,周洋,况燕. SPHK1过表达慢病毒载体的构建和稳定转染SKOV3细胞系的建立[J]. 吉林大学学报(医学版), 2025, 51(6): 1709-1716.
[11]	于晓敏,朱清华,王一伦,任淼,刘子嘉,余泳仪,杜元良,刘东慧,郭森,付秀美. 脂肪源性干细胞联合脱细胞支架对坐骨神经损伤大鼠脊神经节的保护作用及其机制[J]. 吉林大学学报(医学版), 2025, 51(6): 1542-1550.
[12]	相萌,徐冰,王培莎,刘思旗,张少华. 外泌体负载的miR-520a-5p对胎鼠宫内生长受限妊娠结局的改善作用及其机制[J]. 吉林大学学报(医学版), 2025, 51(5): 1230-1239.
[13]	牟燕鸿,李英娜,刘建增,罗春红,孙立伟,姜锐. 防脱育发液对毛发生长的促进作用及其机制[J]. 吉林大学学报(医学版), 2025, 51(5): 1240-1250.
[14]	秦迪,黄丽红,郑庆霜,孙静静,徐为民. 老年肌少症肥胖伴糖尿病患者血清中促炎因子与肌肉质量的关联性分析[J]. 吉林大学学报(医学版), 2025, 51(5): 1293-1302.
[15]	刘建,邢鹭,兰天野,姚璠,王雯,董玉福,吴金普,毕然,孙立伟,陈雪楠,赵为民. 人参醇提取物对老年果蝇睡眠的改善作用及其机制[J]. 吉林大学学报(医学版), 2025, 51(4): 896-903.