Inhibition of proliferative vitreo retinopathy by controling the related transcription factors.

目的.以往的研究表明视网膜色素上皮细胞的增殖和迁移与增生性玻璃体视网膜病变（PVR）的发病机制密切相关。近年来有报道称，诱骗物（decoy），即双链硫代磷酸寡核苷酸，具有与转录因子结合位点相同的序列。在这项研究中，我们测试了E2 F诱饵和NF κ B诱饵在培养的人视网膜色素上皮（RPE）细胞和体内PVR动物模型的增殖中的作用。通过将HVJ脂质体的磷脂酰丝氨酸（Hangai等，1996）替换为DC胆固醇来制备HVJ阳离子脂质体。结果结果表明，E2 F诱饵与人成纤维细胞的核提取物高度结合，NF κ B诱饵也与白细胞介素1 β（IL-1 β）刺激的人RPE细胞的核提取物结合。RT-PCR结果表明，E2 F诱骗基因导入人成纤维细胞后，细胞周期调控因子的表达降低，NF κ B诱骗基因导入人RPE细胞后，IL-1 β刺激的RPE细胞中IL-1 β的转录水平降低。BrdU标记指数和DNA合成（H-胸苷摄取）阐明了E2 F诱饵高度抑制细胞增殖。与乱序诱饵相比，具有NF κ B诱饵的培养皿上的细胞显著抑制细胞迁移。体内实验将HVJ阳离子脂质体介导的NF-κ B诱骗物转染到体外培养的人成纤维细胞中，注射到兔PVR模型的玻璃体腔内。根据Blumenkraz等人的分类对兔PVR的出现进行评分。使用NF κ B诱饵和乱序诱饵的PVR发生率没有显著差异。结论.这些结果表明E2 F和NF κ B诱骗物在体外具有治疗增殖性玻璃体视网膜病变的潜力，其体外应用需要更多的研究。

Purpose. Previous studies suggested that proliferation and migration of retinal pigment epithelial cells are deeply involved in pathogenesis of proliferative vitreo retinopathy (PVR). Recently it is reported that decoys, double-stranded phosphorothioate oligonucleotides, have the same sequence of binding site of transcription factors. In this study we tested the effect of E2F decoys and NFkappaB decoys in the proliferation of cultured human retinal pigment epithelial (RPE) cells and in vivo animal model of PVR.Methods. HVJ cationic liposomes were prepared by replacing phosphatidylserine of HVJ liposomes (Hangai, et al, 1996) to DC cholesterol. Results. It was shown that E2F decoys highly combined to nuclear extracts from human fibroblast cells and NFkappaB decoys also combined to nuclear extracts from human RPE cells stimulated by interleukin 1beta (IL-1beta). RT-PCR indicated that E2F decoys introduced into human fibroblast decreased expression of important factor that is regulating cell cycle, it showed that NFkappaB decoys introduced into human RPE cells reduced transcripts of IL-1beta in IL-1beta stimulated RPE cells. BrdU labeling index and DNA synthesis (H-thymidine uptake) clarified that cell proliferation was highly inhibited with E2F decoys. Cells on culture dishes with NFkappaB decoys were significantly inhibited cell migrations compared with scrambled decoys. In vivo experiment NFkappaB decoys, transferred by HVJ cationic liposomes into human cultured fibroblast, were injected in the vitreous cavity of rabbit PVR model. The appearances of rabbit PVR were scored by a classification of Blumenkraz et al. The incidences of PVR were not significantly different between with NFkappaB decoys and with scrambled decoys. Conclusions. These results suggest that E2F decoys and NFkappaB decoys have the potential to treat proliferative vitreoretinopathy in vitro and it is needed more study to vitro use.