sIL-1R基因重组质粒在山羊颞下颌关节内的表达

目的:观察脂质体携带的可溶性白介素-1受体(sIL-1RI)基因重组质粒注入山羊颞下颌关节(TMJ)上腔后,在TMJ髁突软骨及滑膜组织中的表达,以及在全身其他重要器官组织中的表达,为施行基因治疗TMJ骨关节病(OA)奠定基础。方法:将脂质体携带的sIL-1RI基因重组质粒分别注入6只山羊TMJ的关节上腔。注射后第2、4周,分别处死3只动物。抽取TMJ关节滑液,用ELISA法检测关节滑液中的蛋白表达量。采用q检验,对实验数据进行方差分析和均数间的两两比较。切取完整关节标本以及肝脏、心脏、肾脏和脑组织,免疫组化法检测重组质粒在这些脏器组织中的表达情况。结果:重组质粒pcDNA3/sIL-1RI在山羊TMJ的滑膜细胞、软骨细胞及软骨下组织均有表达。ELISA方法检测结果显示,实验组表达量明显高于对照组(P<0.05)。在山羊肝脏、心脏、肾脏和脑等全身重要组织器官中未见表达。结论:脂质体携带的重组质粒pcDNA3/sIL-1RI,可在山羊TMJ有效地表达,关节外组织未见表达,因此是安全、可行的。

Direct liposomes-mediated soluble interleukin-1 receptor gene delivery in goat temporomandibular joint

PURPOSE:This study is to confirm that soluble interleukin-1receptor(sIL-1RI)gene can be delivered to articular chondrocytes and synovium in goat temporomandibular joint(TMJ)using direct in vivo gene transfer method by liposome.METHODS:Recombiant liposomes vector carrying sIL-1RI gene was injected into the upper joint cavities of both TMJs of six male goats.At2and4weeks after injection,6joints were dissected respectively and the expression of delivered sIL-1RI gene in synovium and articular cartilage of goat TMJ was examined immunohiso-chemically.Meanwhile,the expression of sIL-1RI gene in liver,heart,kidney,and brain of goat was examined immunohisochemically.The protein expression products in TMJ synovial fluid samples were detected by ELISA and statistical significance was assessed using q test.RESULTS:Clear expression of sIL-1RI gene was observed in the synovial membrane,articular cartilage,and subchondral bone tissue of the goat TMJ,while no expression was detected in goat liver,heart,kidney,and brain.The protein expression products could be detected in synovial fluid samples by ELISA,and the level of expression in experimental group were significantly higher than that in the control groups(P<0.05). CONCLUSION:Direct genesurface and articular cartilage of the goat TMJ using the liposomes vector carrying sIL-1RI gene is feasible as an effective in vivo method.