局部使用作用于肿瘤坏死因子α的反义寡核苷酸对实验性小鼠单疱病毒性脉络膜视网膜炎的影响

目的 观察局部使用作用于肿瘤坏死因子α（TNF-α）的反义寡核苷酸对实验性单疱病毒性脉络膜视网膜炎病理过程的影响。 方法 将50只BALB/c小鼠随机分为实验组和对照组，每组25只。将单纯疱疹病毒-Ⅰ型（HSV-Ⅰ）注入所有小鼠右眼（注射眼）前房内，建立单疱病毒性脉络膜视网膜炎的小鼠模型。实验组中分别于HSV-Ⅰ型感染前1 d，感染后1、4 d，将异硫氰酸荧光素标记的作用于TNF-α的反义寡核苷酸2 μl注射于感染小鼠的左眼（非注射眼）球结膜下;对照组则于相同的时间段内注射等量的磷酸盐缓冲液于感染小鼠的左眼（非注射眼）球结膜下，观察两组小鼠眼部的炎症改变并根据有无前房炎症反应、瞳孔和虹膜血管扩张、白内障形成、玻璃体混浊等行临床评分。于病毒感染后10 d处死所有小鼠，观察其组织学变化并采用酶联免疫吸附试验（ELISA）测定小鼠脉络膜视网膜的TNF α含量。 结果 感染后，两组小鼠右眼均表现为急性炎症改变;实验组小鼠左眼炎症反应临床评分明显小于对照组。组织学观察结果:对照组有12只小鼠左眼表现程度不等的坏死性脉络膜视网膜炎，实验组有2只小鼠左眼表现为轻度的脉络膜视网膜炎，两组间视网膜、脉络膜及睫状体部位炎性细胞计数差异有统计学意义（P＜0.05），而前房、玻璃体腔及虹膜部位无明显差别（P＞0.05）。ELISA测定结果:实验组脉络膜视网膜TNF-α含量为（60±1.25） pg，对照组脉络膜视网膜TNF-α含量为（305±1.03）pg，两者相比差异有统计学意义（P＜0.05）。 结论 采用作用于TNF-α的反义寡核苷酸局部治疗小鼠单疱病毒性脉络膜视网膜炎，能明显降低感染小鼠眼内细胞因子TNF-α的含量，减轻眼内脉络膜视网膜炎症反应。 (中华眼底病杂志, 2006, 22: 245-248)

Effect of topical treatment with antisense oligonucleotides targeting tumor necrosis factor-α on experimental HSV-1-induced chorioretinitis in mice

Objective To investigate the effect of topical treatment with antisense oligonucleotides（ASON）targeting tumor necrosis factor-α（TNF-α）on the pathological process of experimental herpes simplex virus type-Ⅰ（HSV-Ⅰ）induced chorioretinitis in mouse eye. Methods Fifty BALB/c mice were randomly divided into experimental and control group（twenty five mice in each group）.HSV chorioretinitis model was induced in each mouse by inoculating 1×105 plaque-forming units （pfu） of HSV-Ⅰ（KOS strain）into anterior chamber of the right eye.In experimental group，Fluorescein isothiocyanate （FITC）-labeled ASON targeting TNF-α 2 μl were injected sub-conjunctiva in the left eye1day before and 1 and 4 days after the infection;while phosphate buffer solution was injected in the same way in control group.The inflammation changes of the eyes in the 2 groups were observed and the clinical grades were assessed according to the extends of anterior-chamber inflammation，vasodilatation of cornea and iris，formation of cataract，and vitreous opacity. All of the mice were executed 10 days after the infection and were observed histologically. The contents of TNF-α in retina and choroid were measured by enzyme-linked immunobsorbent assay（ELISA）. Results After the infection，acute inflammation appeared in the right eyes in both groups. The inflammation of the left eyes in experimental group was significantly milder than which in the control group.Twelve left eyes had necrotic chorioretinitis in different degrees in the control group while 2 left eyes had mild chorioretinitis in the experimental group. The difference of the number of inflammatory cells between the 2 groups was statistically significant in retina，choroid，and ciliary body（P＜0.05）and was not obvious in anterior chamber，vitreous cavity，and iris（P＞0.05）.The content of TNF-α in choroid and retina was（60±1.25）pg in the experimental group and（305±1.03）pg in the control group（P＜0.05）. Conclusions TNF-α ASON treating HSV-Ⅰinduced chorioretinitis may reduce the content of TNF-α in affected mice eyes and decrease the inflammatory reaction. (Chin J Ocul Fundus Dis, 2006, 22: 245-248)