AAV-hTGFβ_1体外转染鼠椎间盘再移植的实验研究  

EXPERIMENTAL STUDY OF RETRANSPLANTATION OF IN VITRO AAV-hTGF β_1-TRANSFECTED CELLS OF INTERVERTEBRAL DISC IN RATS

作　　者：李涛[1] 刘勇[1] 郑文艳[2] 

机构地区：[1]青岛大学医学院附属医院脊柱外科,山东青岛266003 [2]青岛大学医学院附属医院手术室,山东青岛266003

出　　处：《齐鲁医学杂志》2011年第4期325-327,330,共4页Medical Journal of Qilu

摘　　要：目的探讨腺相关病毒（AAV）介导人转化生长因子β1（hTGFβ1）基因体外转染大鼠椎间盘细胞后再移植至大鼠的椎间盘中,对椎间盘高度、Ⅱ型胶原和蛋白聚糖的影响。方法 Wistar大鼠20只,以L6-S1椎间盘作为空白对照组,不进行处理;L5-L6椎间盘作为针刺组,暴露以后用显微注射器从椎间盘的前外侧向椎间盘内注射磷酸盐缓冲液;L4-L5椎间盘作为实验组,手术暴露后用显微注射器从椎间盘的前外侧向椎间盘内注射AAV-hTGFβ1体外转染后的椎间盘细胞悬液。术前、术后4周对大鼠腰椎间盘进行X线检查,术后4周利用反转录-聚合酶链反应（RT-PCR）对腰椎间盘组织中的Ⅱ型胶原和蛋白聚糖的基因表达进行半定量分析。结果针刺组手术前后椎间盘高度系数的变化（%DHI）均低于实验组和空白对照组,差异有显著性（F=321.22,q=30.44、32.56,P〈0.05）;实验组手术前后%DHI略低于空白对照组,差异无显著性（P〉0.05）。针刺组Ⅱ型胶原和蛋白聚糖表达明显低于实验组和空白对照组,差异有显著性（F=55.05、34.79,q=8.96~12.43,P〈0.05）;实验组Ⅱ型胶原和蛋白聚糖表达与空白对照组比较,差异无显著性（P〉0.05）。结论 AAV-hTGFβ1基因体外转染大鼠椎间盘细胞后再移植至大鼠的椎间盘中,有助于椎间盘高度的维持并能提高椎间盘细胞Ⅱ型胶原和蛋白聚糖表达。Objective To study the reimplantation of in vitro AAV-hTGFβ1-transfected cells of intervertebral disc（IVD）,and observe its effect on height of the disc,type Ⅱcollagen and proteoglycan in rats. Methods Twenty Wistar rats were included in this study： L6-S1 IVD was assigned as control group,no managements were done;L5-L6 IVD as puncture group injected with 0.01 mol/L PBS;L4-L5 as experimental group,injected with cell suspension of AAV-hTGFβ1-transfected disc cells.X-ray examination of lumbar IVD of the rats were undertaken before and four weeks after the procedures,and,four weeks later,gene expressions of typeⅡcollagen and proteoglycan were detected by reverse transcription polymerase chain reaction（RT-PCR） and analyzed by semi-quantitative technique. Results The percentage of DHIs in puncture group were remarkably decreased compared with that in experimental group and control group（F=321.22;q=30.44,32.56;P〈0.05）,and the difference of that between experimental group and control group was not significant（P〉0.05）.The expressions of type Ⅱ collagen and proteoglycan in puncture group were weaker than that in experimental group and control group（F=55.05,34.79;q=8.96-12.43;P〈0.05）,and the difference of that between experimental group and control group was not significant（P〉0.05）. Conclusion Retransplantation of AAV-hTGFβ1-transfected disc cells into the intervertebral discs in rats conduces to maintenance of the height of intervertebral disc and enhancement of the expressions of type Ⅱ collagen and proteoglycan.

关 键 词：转化生长因子β 依赖病毒 胶原Ⅱ型 蛋白聚糖 椎间盘 

分 类 号：R681.5[医药卫生—骨科学]