目的：观察NF-κB反义RNA基因在超声联合微泡作用下对类风湿性关节炎（Rheumatoid arthritis，RA）大鼠血清TNF-α、IL-1β水平的影响，探讨其对RA治疗的可行性。材料与方法：采用层-层吸附法将NF-κB反义RNA基因和多聚赖氨酸（Polylysine，PLL）分层吸附在自制的脂质超声微泡造影剂上，制备成载（PLL+DNA+PLL+DNA）的微泡。60只正常清洁级Wister雌性大鼠，分为6组（其中B~F组为造模成功后的RA大鼠），每组10只（20个膝关节）。A组：正常对照组；B组：RA大鼠模型对照组；C组：单纯NF-κB反义RNA基因组；D组：超声+NF-κB反义RNA基因组；E组：微泡+NF-κB反义RNA基因组；F组：超声+微泡+NF-κB反义RNA基因组。ELISA检测比较各组大鼠血清中的细胞因子TNF-α和IL-1β水平变化。结果：模型对照组大鼠血清TNF-α、IL-1β水平明显高于正常大鼠组（P＜0.01），单纯基因组、超声+基因组、微泡+基因组、超声+微泡+基因组大鼠血清TNF-α、IL-1β水平均低于模型对照组（P＜0.05，P＜0.01），单纯基因组、超声+基因组、微泡+基因组3组之间比较无显著性差异（P＞0.05），超声+微泡+基因组与单纯基因组、超声+基因组、微泡+基因组比较有显著性差异（P＜0.05）。结论：NF-κB反义RNA基因在超声联合微泡作用下对RA大鼠血清TNF-α、IL-1β水平有明显的下调作用，有助于提高RA的治疗效果。
Abstract
Objective: To investigate the effect of NF-κB antisense RNA gene delivery mediated by ultrasound-mediated microbubble destruction on serum levels of TNF-α, IL-1β in rheumatoid arthritis(RA) rats and its feasibility for RA therapy. Methods: The layer-by-layer assembly technique was applied to attach multiple layers of gene and polyllysine onto prepared lipid-coated microbubbles(PLL+DNA+PLL Apply+DNA)． Sixty rats were divided into six groups(10 rats with 20 knees in each group). Group A: normal control group; group B: blank control group of RA rats; group C: NF-κB antisense RNA gene plasmid group; group D: ultrasound+NF-κB antisense RNA gene plasmid group; group E: microbubbles+NF-κB antisense RNA gene plasmid group; group F: ultrasound+microbubbles+NF-κB antisense RNA gene plasmid group. The serum levels of TNF-α, IL-1β in RA rats were detected with ELISA. Results: The levels of serum TNF-α and IL-1β were obviously higher than normal control group. Those of group C to F were notably lower than that of the RA control group(P＜0.05, P＜0.01). There was no statistical difference in the levels of serum TNF-α and IL-1β among group C, D and E. But there was statistics difference between group F and group C to E(P＞0.05). Conclusion: NF-κB antisense RNA gene transfection efficiency on RA rat’s joints synovial tissues can be enhanced by ultrasound-mediated microbubble destruction, which can down-regulate the levels of serum TNF-α and IL-1β, and enhance the therapeutic effect of RA.
关键词
关节炎 /
类风湿 /
白细胞介素1 /
肿瘤坏死因子&alpha /
超声检查 /
介入性
Key words
Arthritis, rheumatoid /
Interleukin-1 /
Tumor necrosis factor-alpha /
Ultrasonography, interventional
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基金
国家自然科学基金（30860270）；海南省自然科学基金（30854，813208）。
