Yovandich J, O'Malley B, Sikes M, Ledley F D
Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030, USA.
Hum Gene Ther. 1995 May;6(5):603-10. doi: 10.1089/hum.1995.6.5-603.
We studied reporter gene expression in synovial tissue after intra-articular administration of an expression plasmid into the knees of rabbits and rats. In both species, administration of a plasmid encoding beta-galactosidase led to gene expression in the synovial cells lining the joint. Expression correlated with the presence of plasmid DNA in synovial tissue extracts. Studies with a plasmid encoding chloramphenicol acetyltransferase demonstrated that gene expression persists for 2-5 days after administration. Southern blotting demonstrated that the administered plasmid was taken up rapidly by synovial tissue and degraded. By 24 hr after administration, no intact plasmid could be detected by Southern blotting, although small amounts of plasmid could be amplified by PCR up to 7 days. Administration of a plasmid encoding human growth hormone demonstrated that this product could be expressed from synovial cells and secreted into the synovial fluid. The histological distribution of gene expression in synovium resembles the known distribution of particulate materials injected into the joint and suggests that plasmid DNA is taken up by nonspecific endocytosis like other particulate materials during the remodeling of synovial fluid.
我们研究了将表达质粒关节腔内注射到兔和大鼠膝关节后，滑膜组织中的报告基因表达情况。在这两个物种中，注射编码β-半乳糖苷酶的质粒均导致关节内衬滑膜细胞中的基因表达。表达与滑膜组织提取物中质粒DNA的存在相关。对编码氯霉素乙酰转移酶的质粒的研究表明，给药后基因表达持续2至5天。Southern印迹法显示，给药的质粒被滑膜组织迅速摄取并降解。给药后24小时，Southern印迹法检测不到完整质粒，尽管少量质粒在7天内可通过PCR扩增。注射编码人生长激素的质粒表明，该产物可由滑膜细胞表达并分泌到滑液中。滑膜中基因表达的组织学分布类似于注入关节的颗粒物质的已知分布，这表明在滑液重塑过程中，质粒DNA像其他颗粒物质一样通过非特异性内吞作用被摄取。
