转GGCX基因治疗兔骨性关节炎的体外实验研究

发布时间：2018-06-07 06:51

本文选题：软骨 + 关节　；参考：《重庆医学》2017年04期

【摘要】：目的探索GGCX基因对兔骨性关节炎软骨细胞MMP13的影响及其在兔骨性关节炎软骨退变中的作用。方法取体质量(2.0±0.2)kg日本大耳兔6只,随机分为3组,每组设立一只对照兔,分别在2、4、6周使用膝前交叉韧带切断术构建模型,6周后模型兔构建成功,分离关节软骨,经消化分离后将软骨细胞接种到6孔细胞培养板,将软骨细胞分为空白组、阴性对照组(转染不含GGCX基因慢病毒)、转染组(转染含GGCX基因慢病毒)3组。转染组则均以LipofectamineTM2000为转染媒介。用PCR法及蛋白免疫印迹法(Western blot)检测GGCX和MMP13的表达。结果转染组GGCX有较高的表达水平;转染组MMP13的表达水平降低,且二者与空白组及阴性对照组比较,差异均有统计学意义(P0.05)。结论 GGCX基因过表达可明显降低其软骨细胞MMP13的表达,为将基因治疗运用于体外骨性关节炎提供实验基础。
[Abstract]:Objective to investigate the effect of GGCX gene on MMP13 in rabbit osteoarthritis chondrocytes and its role in osteoarthritis cartilage degeneration. Methods six Japanese large ear rabbits with 2.0 卤0.2)kg body mass were randomly divided into 3 groups. Each group was divided into 3 groups, each group was divided into 3 groups. The model was constructed 6 weeks after the operation of anterior cruciate ligament (ACL), and the articular cartilage was separated. After digestion and isolation, chondrocytes were inoculated into 6-well cell culture plate. Chondrocytes were divided into blank group, negative control group (transfection without GGCX gene lentivirus) and transfection group (transfected with GGCX gene lentivirus). In the transfection group, LipofectamineTM2000 was used as the transfection medium. The expression of GGCX and MMP13 was detected by PCR and Western blotting. Results the expression of GGCX in the transfected group was higher than that in the control group, and the expression level of MMP13 in the transfected group was lower than that in the blank group and the negative control group (P 0.05). Conclusion overexpression of GGCX gene can significantly reduce the expression of MMP13 in chondrocytes and provide experimental basis for the application of gene therapy in osteoarthritis in vitro.
【作者单位】：南昌大学第二附属医院骨科;
【基金】：国家自然基金资助项目(81301561)
【分类号】：R684.3

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