基因沉默与诱导双向治疗类风湿性关节炎-外科学(骨科)专业论文.docxVIP

万方数据 万方数据 体外观见各组的关节修复没有明显差别。 结论： UTMD 联合转染试剂 Entranster 应用可以作为一种新的转染方法，提高基因转染 效率；两种方法联合进行体内转染后，亦能够在体内表达，从而达到治疗的目的。 【关键词】 类风湿性关节炎；环氧合酶-2；聚蛋白多糖酶-1；人类胰岛素生长 因子；基因治疗 硕士研究生:姜岩 指导老师: 张海宁副教授 Using Gene Transfection of Gene Silencing and Induction to Treat the Rheumatoid Arthritis Abstract Objection:1. To investigate the efficiency of aggrecanase-1 shRNA transfection on MSCs mediated by ultrasound-targeted microbubble and Entranster；2. Establishment of the rheumatoid arthritis model of the rabbit and evaluation; 3. To explore the therapeutic effect of aggrecanase-1 shRNA，COX-2 shRNA, hIGF transfection mediated by ultrasound-targeted microbubble and Entranster in the rheumatoid arthritis model of rabbits. Method: According to the standard bone marrow puncture procedure, the marrow was aspirated to culture the human bone marrow mesenchymal stem cells (hBMSCs). hBMSCs were cultured and passaged to third generation. Then the third generation cells were cultured in 24-well plates for 24 hours, and divided into four groups: (a) plasmid + ultrasound-targeted microbubble destruction group (UTMD), (b) plasmid + transfection reagent Entranster group (ENTR), (c) plasmid + ultrasound-targeted microbubble destruction group + transfection reagent Entranster group (UTMD+ENTR), (d) plasmid（CONTR）. To observe the fluorescence of every group on day 2, 4, 6 by the inverted fluorescence microscope; To detect the mRNA expression of aggrecanase-1 by RT-PCR. The establishment of the rheumatoid arthritis model of the rabbit: Mix the bovine type II collagen solution with the complete Freunds adjuvant into emulsion. Then continuous subcutaneous injection of 1ml emulsion at 10 points of scapular area. To inject 1ml collagen solution into knee joint to sensitize RA group rabbits. And then After operative, we evaluated the model through the degree of joint swelling, the grade of the arthritis and pathohistological examitation. In vivo transfection experiments: The successful rheumatoid arthritis model