髓核内注射骨髓间质干细胞对椎间盘基质的影响  

The effect of injecting bone mesenchymal stem cells into nucleus pulposus on the biochemistry of intervertebral disc

作　　者：张银刚[1] 郭雄[2] 李天清[1] 王世捷[2] 李曙明[1] 许鹏[2] 任峰玲[2] 

机构地区：[1]西安交通大学医学院第一附属医院骨科,陕西西安710061 [2]西安交通大学医学院环境与疾病相关基因教育部重点实验室,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2007年第6期675-679,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30400163)

摘　　要：目的研究髓核内注射骨髓间质干细胞对椎间盘基质的影响。方法32只新西兰大白兔,L2-3、L3-4、L4-5、L5-6被随机分为正常对照组、生理盐水对照组、细胞移植组Ⅰ、细胞移植组Ⅱ(BMP2转基因组)。髓核内注射后1、3、6月时间点,用Ⅰ、Ⅱ型胶原单克隆抗体免疫组化染色分析胶原表型的改变,用实时荧光定量PCR分析Ⅰ、Ⅱ型胶原及蛋白多糖的mRNA表达,采用重复测量方差分析进行统计学分析。结果术后1、3、6月时间点,细胞移植组髓核有较多的细胞,蛋白多糖染色显示细胞移植组髓核兰染增加,免疫组化显示Ⅰ型胶原实验各组未见明显差异,Ⅱ型胶原细胞移植组有较强的红染颗粒沉积。实时荧光定量RT-PCR分析显示,在细胞移植组,蛋白多糖和Ⅱ型胶原的mRNA增加,而Ⅰ型胶原mRNA则无改变。与细胞移植组Ⅰ比较,细胞移植组Ⅱ蛋白多糖和Ⅱ型胶原mRNA增加。结论骨髓间质干细胞移植能够提高髓核中蛋白多糖和Ⅱ型胶原的含量,复合BMP2转基因有"分子增强效应",提示基于骨髓间质干细胞移植策略是治疗椎间盘退变潜在的有效方法。Objective To explore the effect of injecting bone mesenchymal stem cells into nucleus pulposus on the biochemistry of intervertebral disc. Methods Totally 32 rabbits were used, each of whose lumbar discs from the second to fifth were randomly divided into normal group, saline group, cell transplantation group I , and cell transplantation group Ⅱ（group of the transferring gene coding BMP）. At 1 month-, 3 month-, 6 month-time point after operation, immunohistochemistry was used to determine the change of phenotype of collagen, real time PCR was used to analyze the expression of collagen type Ⅰ and Ⅱ and proteoglycan, followed by repeated ANOVA used to analyze the data. Results At 1 month-, 3 month-, 6 month-time point after operation, there were more cells in cell transplantation group than in normal group or saline group. Toludine blue staining showed that there was deeper blue in cell transplantation group than in normal group or saline group. Immunohistochemistry showed there was deeper red in cell transplantation group than in normal group or saline group in collagen type Ⅱ , whereas there was no change in collagen Ⅰ among the groups. Real time PCR showed that the expression of proteoglycan or collagen type Ⅱ was increased in cell transplantation group whereas collagen type Ⅰ remained unchanged among the groups. In addition, compared to that of cell transplantation group Ⅰ , the expression of proteoglycan or collagen type Ⅱ was increased in cell transplantation group II ~ Conclusion The transplanted bone mesenchymal stem cells can increase the amount of proteoglycan or collagen type Ⅱ; combined with the transfer of BMP2, the effect is overlaying, suggesting that the therapeutic strategy based on bone mesenchymal stem cells may be effective in prevention and treatment of intervertebral disc degeneration.

关 键 词：骨髓间质干细胞 椎间盘 基因治疗 

分 类 号：R68[医药卫生—骨科学]