角膜穿通伤后炎症因子的表达特征及其在房水中相对含量的动态变化

Expression of inflammatory factors and dynamic changes of their related contents in aqueous humor after corneal penetrating wound

目的研究角膜穿通伤后炎症因子在眼球壁的分布特征及其在房水中相对含量的动态变化,初步探讨角膜穿通伤后眼内炎的发病机制。方法清洁级成年雌性SD(Sprague Dawley)大鼠50只,随机分为正常对照组和角膜穿通伤后6h、24h、48h、72h组,每组各10只。角膜穿通伤组大鼠制作角膜穿通伤模型,分别用免疫荧光染色法检测损伤后各时间点眼球壁组织中热休克蛋白90(heat shock protein90,HSP90)、肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)、白细胞介素(interleukin,IL)-1β、IL-10的组织定位;并用免疫印迹法分别检测损伤后各时间点房水中上述因子相对含量的动态变化。结果 HSP90、IL-10、TNF-α和IL-1β在正常房水和眼球组织中含量较少。HSP90于损伤后6h在房水中含量达到高峰,与其他各组相比差异均有统计学意义(均为P<0.05),随后逐渐减少,免疫荧光染色结果显示其主要表达于视网膜视锥和视杆细胞层;TNF-α、IL-1β于损伤后24h在房水中达到高峰,与其他各组相比差异均有统计学意义(均为P<0.05),之后逐渐减少,主要表达于角膜、睫状体、视网膜;IL-10于损伤后72h在房水中含量达到高峰,与其他各组相比差异均有统计学意义(均为P<0.05),主要表达于角膜和视网膜。结论角膜穿通伤后应激反应可促使眼球局部炎症因子的表达,导致继发性炎症损伤。角膜穿通伤后应早期局部使用免疫抑制剂抑制炎症反应,保护眼球壁组织细胞。

Objective To investigate distributions of inflammatory factors in the eyeball walls and dynamic changes of their related contents in aqueous humor for probing the pathogenesis of endophthalmitis after corneal penetrating wound. Methods Fifty clearing adult female Sprague Dawley rats were randomly divided into control group and 6 hours, 24 hours, 48 hours and 72 hours after corneal penetrating wound groups, 10 rats in each group. Rat models with corneal penetrating wound were made. Immunofluorescence staining was used to detect location of heat shock protein 90 （ HSP90 ）, tumor necrosis factor -a （ TNF-a ）, interleukin- 1 β （ IL- 1β ） and IL- 10 in eyeball tissues at different time points. Immunoblotting was used to detect dynamic changes of relative contents of these factors in aqueous humor at different time points after wound. Results The related contents of HSP90, IL-10, TNF-a, and IL-1β were at low level in normal eyeball wall and aqueous humor. HSP90 in aqueous humor went to peak at 6 hours after corneal penetrating wound, which was statistically significant compared with other groups （ all P 〈 0.05 ）, and then gradually reduced. Immunofluorescence results showed that HSP90 mainly expressed in the retinal cone cell layer and retinal rod cell layer;TNF-a and IL-1β reached their peaks in aqueous humor at 24 hours after the wound, which were statistically significant compared with other groups （ all P 〈 0.05 ）. then gradually decreased, and they mainly expressed in cornea, ciliary body and retina; IL-10 reached its peak in aqueous humor at 72 hours after the wound,which was statistically significant compared with other groups （ all P 〈 0.05 ）, which could be seen mainly in cornea and retina. Conclusions After corneal penetrating wound, stress reaction can promote the expression of inflammatory factors in eyeball,leading a secondary inflammatory injury. We should use immtmosuppressive drugs to inhibit the local inflammatory response for protecting eyeball wall tissue cells at early stage after corneal penetrating wound.