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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-型(HSV-)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-型感染前1d,感染后1、4d,将异硫氰酸荧光素标记的作用于TNF-α的反义寡核苷酸2μl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10d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。
Objective To investigate the effect of topical antisense oligonucleotide on tumor necrosis factor α (TNF-α) in the pathogenesis of experimental herpes simplex virus chorioretinitis. Methods 50 BALB / c mice were randomly divided into experimental group and control group, with 25 rats in each group. Herpes simplex virus-type (HSV-) was injected into the anterior chamber of all mice right eye (injection eye) to establish a mouse model of herpes simplex virus chorioretinitis. In the experimental group, 2 μl of antisense oligonucleotide labeled with fluorescein isothiocyanate labeled TNF-α was injected into the left eye of non-infected mice at 1 day before HSV-infection and 1 and 4 days after infection respectively Injected into the subconjunctival bulb). In the control group, the same amount of phosphate buffered saline (PBS) was injected into the conjunctiva of the left eye (non-injected eye) of the infected mice in the same time period to observe the inflammatory changes in the eyes of the two groups And based on the presence or absence of anterior chamber inflammation, pupil and iris vasodilation, cataract formation, vitreous opacity and other clinical scores. All mice were killed 10 days after virus infection, histological changes were observed and the content of TNF-α in the chorio-retina of mice was determined by enzyme-linked immunosorbent assay (ELISA). Results After infection, the right eye of both groups showed acute inflammatory changes. The clinical scores of left eye inflammatory reaction in the experimental group were significantly less than those in the control group. Histological Observation Results Twelve mice in the control group showed necrotizing chorioretinitis with varying degrees of left eye performance. Two of the mice in the experimental group showed mild chorioretinitis on the left eye. Retinal, choroid and eyelashes There were significant differences in inflammatory cell counts between the body parts (P <0.05), but there was no significant difference between the anterior chamber, the vitreous cavity and the iris (P> 0.05). The results of ELISA showed that the content of TNF-α in the choroidal retina was (60 ± 1.25) pg in the experimental group and (305 ± 1.03) pg in the choroidal retina in the control group, with statistical significance (P <0.05). CONCLUSION: Topical treatment of mouse herpes simplex virus chorioretinitis with antisense oligonucleotide acting on TNF-α can significantly reduce the content of intracellular TNF-α in the infected mice and reduce the intraocular chorioretinal inflammation.