MyD88依赖性NF-κB信号途径改变对家兔PVR模型玻璃体内TNF-α变化的影响

MyD88-dependent NF-κB signaling pathways on changes of TNF-α in vitreous body of rabbit PVR models

目的以有色家兔为实验动物制作兔眼PVR模型,并向眼内转染腺病毒携带的无功能-MyD88（Dn-MyD88）,通过阻断核转录因子-κB（nuclear factor kappa B,NF-κB）信号通路而观察增生性玻璃体视网膜病变（proliferative vitreoretinopathy,PVR）中肿瘤坏死因子-α（tumor necrosis facturα,TNF-α）含量的变化。方法制备含血小板浓度为（50~100）×109L的血浆,制作兔眼PVR模型,向兔眼玻璃体内转染腺病毒携带的Dn-MyD88,运用Western blotting检测向玻璃体内转染腺病毒携带的无功能MyD88成功,并于注射后第1天、第7天、第14天、第21天和第28天分别处死动物,在液氮低温条件下,取得玻璃体样本,对玻璃体液行酶联免疫双抗夹心法（ELISA）检测玻璃体中TNF-α含量。结果空白对照组各时间点玻璃体中TNF-α的含量并无明显变化,PVR注射生理盐水组各时间点玻璃体中TNF-α的含量随着时间的推移呈上升趋势,在造模后第28天时达到最高,为（1073.74±190.43）ng.L-1,PVR注射腺病毒携带的Dn-MyD88组各时间点玻璃体中TNF-α含量呈缓慢上升趋势,在造模后第28天时TNF-α的含量为（612.47±59.29）ng.L-1,与PVR注射生理盐水组相比,差异有统计学意义（P〈0.05）。结论在兔眼PVR模型中通过转染腺病毒携带的Dn-MyD88阻断MyD88依赖性NF-κB信号传导通路而抑制PVR的发生发展,可为PVR的治疗提供新的方法。[眼科新进展2010;30（10）：933-936]

Objective To investigate the changes of tumor necrosis factor α（TNF-α）in proliferative vitreoretinopathy（PVR）by making rabbit PVR models to transfer adenovirus loaded nonfunction MyD88（Dn-MyD88）for inhibiting nuclear factor kappa B（NF-κB）signaling pathways.Methods Blood plasma with thrombocyte concentration of 50×109 to 100×109 was prepared to make PVR models in rabbit eyes.Adenovirus loaded Dn-MyD88 was transferred into vitreous body of rabbit eyes.Western blotting was used to detect adenovirus loaded Dn-MyD88 transferred into vitreous body.Rabbits were respectively sacrificed at the 1st day,7th day,14th day,21st day and 28th day after injection.Samples of vitreous body were acquired under liquid nitrogen hypothermia.Levels of intravitreous TNF-α were detected by ELISA assay.Results Levels of intravitreous TNF-α were unchanged in normal control group at different time points,improved with time in PVR injecting normal sodium group,went to peak as（1 073.74±190.43）ng·L-1 at the 28th day after modeling,improved slowly in PVR injecting adenovirus loaded Dn-MyD88 group at different time points and was（612.47±59.29）ng·L-1 at the 28th day after modeling,which was lower than that in PVR injecting normal sodium group and statistical difference was found（P0.05）.Conclusion With transferring adenovirus loaded Dn-MyD88 can obstruct MyD88 dependent NF-κB signaling pathways to inhibit progress of PVR in rabbit eye PVR models,which provide a new way for PVR treatment.