Jing Xiang-Xiang, Liu Jie, Yang Bing-Ang, Fu Shao-Qing, Wu Tang-Na, Wang Dong-Lin
Department of Medical Ultrasonics, Hainan Provincial People's Hospital, Haikou, Hainan 570311, P.R. China.
Department of Emergency Orthopedics, Hainan Provincial People's Hospital, Haikou, Hainan 570311, P.R. China.
Exp Ther Med. 2014 May;7(5):1396-1402. doi: 10.3892/etm.2014.1579. Epub 2014 Feb 24.
The aim of the present study was to explore the feasibility of enhancing green fluorescent protein (EGFP) gene transfection into the synovial joint tissues of rats with rheumatoid arthritis (RA) by ultrasound-mediated microbubble destruction. An optimal SonoVue dose was determined using 40 normal rats categorized into five groups according to the various doses of microbubbles used. At 1 week after ultrasound irradiation, the rats were sacrificed. Damage to the joint synovial tissues was observed with hematoxylin and eosin histopathological staining under a microscope. A further 44 normal rats were used to establish a rat model of RA, and were then categorized into four groups: EGFP, ultrasound + EGFP, microbubbles + EGFP and ultrasound + microbubbles + EGFP. The last group was irradiated with ultrasound for 10 min following the injection of 300 μl SonoVue and 10 μg EGFP into the joint cavity. Rats were sacrificed after 3 days and synovial tissue was collected from the knee joints for observation of EGFP with fluorescence microscopy and analysis by quantitative polymerase chain reaction. EGFP expression was observed in the synovial tissues of all groups. However, high EGFP expression levels were observed in the ultrasound + microbubbles + EGFP group. No statistically significant differences (P>0.05) were observed in the EGFP expression levels between the EGFP, ultrasound + EGFP and microbubbles + EGFP groups. However, EGFP expression levels in the EGFP, ultrasound + EGFP and microbubbles + EGFP groups significantly differed (P<0.05) from that in the ultrasound + microbubbles + EGFP group. Therefore, ultrasound-mediated microbubble destruction improved EGFP transfection efficiency into the joint synovial tissues of rats with RA.
本研究的目的是探讨通过超声介导的微泡破坏增强绿色荧光蛋白（EGFP）基因转染类风湿性关节炎（RA）大鼠滑膜关节组织的可行性。使用40只正常大鼠，根据所用微泡的不同剂量分为五组，确定最佳声诺维剂量。超声照射1周后，处死大鼠。在显微镜下用苏木精和伊红组织病理学染色观察关节滑膜组织损伤情况。另外44只正常大鼠用于建立RA大鼠模型，然后分为四组：EGFP组、超声+EGFP组、微泡+EGFP组和超声+微泡+EGFP组。最后一组在向关节腔内注射300μl声诺维和10μg EGFP后接受10分钟超声照射。3天后处死大鼠，从膝关节收集滑膜组织，用荧光显微镜观察EGFP并通过定量聚合酶链反应进行分析。所有组的滑膜组织中均观察到EGFP表达。然而，在超声+微泡+EGFP组中观察到较高的EGFP表达水平。EGFP组、超声+EGFP组和微泡+EGFP组之间的EGFP表达水平无统计学显著差异（P>0.05）。然而，EGFP组、超声+EGFP组和微泡+EGFP组的EGFP表达水平与超声+微泡+EGFP组相比有显著差异（P<0.05）。因此，超声介导的微泡破坏提高了EGFP转染RA大鼠关节滑膜组织的效率。
