PDGFR-αASODN玻璃体内注射对视网膜的毒性作用
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摘要
目的:探讨PDGFR-αASODN对视网膜的毒性作用。方法:选择健康成年有色家兔24只,随机分为4组,每组6只;4组兔子3组右眼玻璃体分别注射0.1 m L不同浓度的PDGFR-αASODN/lipofectamine TM 2000溶液,另外1组注射0.1 m L平衡盐溶液作为对照组;4组兔子的左眼不注药。于注药后第1、7、14及28天,对4组兔子的右眼行裂隙灯、间接检眼镜、视网膜电图(ERG)检查;第28天,取4组兔子的眼球,对视网膜组织进行光镜HE、免疫组化及透射电镜的观察。结果:裂隙灯、间接检眼镜检查,各组在各个检查时间点均未发现异常。ERG b波振幅,实验组与对照组比较差异无显著性。注药后第28天,光镜下HE和免疫组化检查,各组视网膜组织均未发现任何病理变化。注药后第28天电镜检查,D组视网膜感光细胞:部分膜盘间隙扩张,部分膜盘融合,少数细胞核周围间隙略增大,其细胞核形态略不规则。结论:玻璃体内注射0.1 m L PDGFR-αASODN/lip2000时,PDGFR-αASODN的浓度≤1.5μmol/L是较为安全的。
Objective To explore the toxic effect of platelet-derived growth factor receptor(PDGFR)-αantisense oligonucleotide(αASODN) on the retina. Methods Twenty-four healthy adult colored rabbits were selected and randomly divided into four groups in six for each group. Intravitreous injections of 0.1ml different density diluents containing PDGFR-α ASODN and liposome were performed in the right eyes in 3 groups. The other group was injected with 0.1 m L balanced salt solution(BSS) as the control group. The left eyes of all animals were not rejected. Slit lamp examination, indirect ophthalmoscopy and electroretinogram(ERG) examination were performed at 1, 7, 14 and 28 days after the injection. On the day 28, the right eyes were harvested, and HE、immunohistochemistry and transmission electron microscopy of retinal tissue were performed. Results The slit lamp, indirect ophthalmoscope examination of all groups were normal in each time. ERG examination yielded no difference in the amplitude of b wave between the treated groups and the normal control group. Pathological changes of the retinal tissue were not observed in the examinations of HE and immunohistochemical at the day 28 after injection. Electron microscope observation of retinal photoreceptor cells in the group D showed the parts of gaps between membranous discs were expanding, parts of were fusing, a few of clearances around cell nucleus were slightly enlarged,and the shapes of the cell nucleus were slightly irregular. Conclusions To inject 0.1 m L PDGFR-αASODN / lip2000 into the vitreous chamber, PDGFR-αASODN can be relatively safe when its concentration is less than 1.5 μmol / L.
Objective To explore the toxic effect of platelet-derived growth factor receptor(PDGFR)-αantisense oligonucleotide(αASODN) on the retina. Methods Twenty-four healthy adult colored rabbits were selected and randomly divided into four groups in six for each group. Intravitreous injections of 0.1ml different density diluents containing PDGFR-α ASODN and liposome were performed in the right eyes in 3 groups. The other group was injected with 0.1 m L balanced salt solution(BSS) as the control group. The left eyes of all animals were not rejected. Slit lamp examination, indirect ophthalmoscopy and electroretinogram(ERG) examination were performed at 1, 7, 14 and 28 days after the injection. On the day 28, the right eyes were harvested, and HE、immunohistochemistry and transmission electron microscopy of retinal tissue were performed. Results The slit lamp, indirect ophthalmoscope examination of all groups were normal in each time. ERG examination yielded no difference in the amplitude of b wave between the treated groups and the normal control group. Pathological changes of the retinal tissue were not observed in the examinations of HE and immunohistochemical at the day 28 after injection. Electron microscope observation of retinal photoreceptor cells in the group D showed the parts of gaps between membranous discs were expanding, parts of were fusing, a few of clearances around cell nucleus were slightly enlarged,and the shapes of the cell nucleus were slightly irregular. Conclusions To inject 0.1 m L PDGFR-αASODN / lip2000 into the vitreous chamber, PDGFR-αASODN can be relatively safe when its concentration is less than 1.5 μmol / L.
引文
[1]孙洁,谢立信.反义基因治疗及其在眼科研究中的应用[J].中华眼科杂志,2005,41(1):87-90
[2]周庆芸,张学东.增生性玻璃体视网膜病变及蛋白质组学研究[J].眼科研究,2010,28(1):93-96..
[3]蒋姣姣,彭燕一,黄海,等.PDGF-α受体反义寡核苷酸治疗实验性增生性玻璃体视网膜病变[J].眼科新进展,2013,33(9):812-815,821.
[4]李新宇,李贵刚,朱雪菲.Bcl-xl/Lip2000比值对人角膜基质细胞转染效率的影响[J].眼科新进展,2006,7(26);524-526.
[5]Akhtar S,Hghes MD,Khan A,et al.The delivery of antisense therapeutics[J].Adv Drug Del Rev,2000,44(1):3-21.
[6]Chaumont CG,Seksek 0,Grzybowska J,et al.Delivery systems for antisense oligonucleotides[J].Pharmacol Ther,2000,87(2-3):255-277.
[7]陈海靓,陈智,梁文权.反义寡核苷酸脂质体复合物性质对细胞摄入行为的影响[J].药学学报,2002,37(9):728-732.
[8]邱梅园,彭燕一,黄岚珍,等.PDGF-α受体反义寡核苷酸对体外视网膜色素上皮细胞增殖和凋亡的影响[J].国际眼科杂志,2011,11(2):229-231.
[9]Kroll J,Waltenberger J.The vascular endothelial growth factor receptor KDR acti-vates multiple signal transduction pathways in porcine aortic endothelial cells[J].J Biol Chem,2004,272(51):32521-32527.
[10]Mori K,Gehlbach P,Ando A,et al.Retina-specific expression of PDGF-B versus PDGF-A:vascular versus nonvascular proliferative retinopathy[J].Invest Ophthalmol Vis Sci,2002,43(6):2001-2006.
[2]周庆芸,张学东.增生性玻璃体视网膜病变及蛋白质组学研究[J].眼科研究,2010,28(1):93-96..
[3]蒋姣姣,彭燕一,黄海,等.PDGF-α受体反义寡核苷酸治疗实验性增生性玻璃体视网膜病变[J].眼科新进展,2013,33(9):812-815,821.
[4]李新宇,李贵刚,朱雪菲.Bcl-xl/Lip2000比值对人角膜基质细胞转染效率的影响[J].眼科新进展,2006,7(26);524-526.
[5]Akhtar S,Hghes MD,Khan A,et al.The delivery of antisense therapeutics[J].Adv Drug Del Rev,2000,44(1):3-21.
[6]Chaumont CG,Seksek 0,Grzybowska J,et al.Delivery systems for antisense oligonucleotides[J].Pharmacol Ther,2000,87(2-3):255-277.
[7]陈海靓,陈智,梁文权.反义寡核苷酸脂质体复合物性质对细胞摄入行为的影响[J].药学学报,2002,37(9):728-732.
[8]邱梅园,彭燕一,黄岚珍,等.PDGF-α受体反义寡核苷酸对体外视网膜色素上皮细胞增殖和凋亡的影响[J].国际眼科杂志,2011,11(2):229-231.
[9]Kroll J,Waltenberger J.The vascular endothelial growth factor receptor KDR acti-vates multiple signal transduction pathways in porcine aortic endothelial cells[J].J Biol Chem,2004,272(51):32521-32527.
[10]Mori K,Gehlbach P,Ando A,et al.Retina-specific expression of PDGF-B versus PDGF-A:vascular versus nonvascular proliferative retinopathy[J].Invest Ophthalmol Vis Sci,2002,43(6):2001-2006.