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(P<0.05)。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。
Objective To research the effect of GGCX gene on MMP13 in rabbit osteoarthritis cartilage cells and investigate its effect on osteoarthritis cartilage degeneration. Methods Six Japanese big ear rabbits weighted （2.0±0.2）kg were randomly divided into three groups,each group seted a rabbit as Control. Anterior cruciate ligament transection method was used to build osteo- arthritis cartilage degeneration model at second, forth, sixth week. Articular cartilage was separated successfully after the model were built,cartilage cells were divided from articular cartilage and cultured in 6-well cell culture plate. Cartilage cells were divided into blank group, neglative control group and transfection group. PCR method and Western blot were conducted to detect GGCX and MMP13 expressed at the level of mRNA and protein. Results Compare with blank group and negative control group, the expressin level of GGCX incresed,while the MMP13 expression level dcresed（P〈0.05）. Conclusion Over expression of GGCX gene can obviously decrease the expression of MMP13,it provide experimental basis for osteoarthritis of the in vitro gene therapy.
出处
《重庆医学》
CAS
北大核心
2017年第4期436-438,共3页
Chongqing medicine
基金
国家自然基金资助项目(81301561)
