体外培养大鼠软骨细胞退变现象的连续观察  

The continuous observation of chondrocyte degeneration in vitro of rats

作　　者：陈晓东[1] 林建华[2] 

机构地区：[1]深圳平乐骨伤科医院,广东深圳518010 [2]福建医科大学附属第一医院骨科

出　　处：《中国老年学杂志》2008年第7期652-655,共4页Chinese Journal of Gerontology

基　　金：福建省科技计划重大资助项目(2004Y018)

摘　　要：目的对体外培养大鼠软骨细胞退变现象进行连续观察,为进一步研究软骨退变机制实验及药物干预实验提供基础资料。方法进行大鼠软骨细胞取材并传代培养,对各代次细胞进行连续性观察,采用倒置相差显微镜、透射电镜观察细胞形态与微结构,免疫组化法检测增殖细胞核抗原(PCNA),阿力新蓝染色检测胞外基质硫酸糖氨多糖(GAG)含量和结构,反转录-聚合酶链式反应方法检测软骨细胞Ⅱ型胶原和Aggrecan表达,观察软骨细胞退变。结果第四代软骨细胞出现长梭形改变,电镜下超微结构出现核固缩或水肿、细胞器减少或变形等退变表现,PCNA表达百分率明显减少(P<0.01),GAG含量、长链分子百分比显著减少(P<0.01),Ⅱ型胶原及Aggrecan mRNA表达显著减少(P<0.01)。结论体外培养软骨细胞第四代出现明显功能退变。Objective To provide the basic data for further studying the chondrocyte degeneration mechanism and its drug intervention experiment by observing continuously chondrocyte degeneration in vitro of rats. Methods The chondrocyte of the rats were sampled to subculture and observe. Inverted phase contrast and transmission electron microscopes were used to observe the cell form and microstructure. Proliferating cell nuclear antigen （PCNA） was detected by immunohistochemisty. Extracellular glycosaminoglycan （GAG） content and structure were determined by alcian blue staining. Chondrocyte Ⅱ type collagen and aggrecan expression were detected by RT-PCR to observe its degeneration. Results The fourth chondrocyte showed long fusiform shape, karyopyknosis or dropsy, decreased or transfigured generation organelle under electron microscope. The expression rate of PCNA, GAG content and the percentage of long chain molecule and the expressions of Ⅱ type collagen and aggrecan mRNA were obviously decreased （all P 〈 0. 01 ）. Conclusions The fourth generation chondrocyte cultured in vitro could have obvious functional degeneration.

关 键 词：软骨细胞 退变 增殖 去分化 

分 类 号：R339.03[医药卫生—人体生理学] Q255[医药卫生—基础医学]