视网膜缺血再灌注损伤大鼠视网膜中白细胞介素-23和白细胞介素-17的表达及意义

Expression and significance of interleukin-23 and interleukin-17 in rat retina after ischemia-reperfusion injury

目的通过建立视网膜缺血再灌注损伤(retinal ischemia-reperfusion injury,RIRI)模型,观察白细胞介素-23(interleukin-23,IL-23)和白细胞介素-17(interleukin-17,IL-17)在Sprague-Dawley(SD)大鼠视网膜上的表达,探讨其在RIRI中的可能作用机制。方法采用随机数字表法将75只健康无眼疾SD大鼠随机分为正常对照组和模型组。正常对照组不作任何处理,模型组采用前房灌注生理盐水升高眼压法建立RIRI模型。分别在建模后12 h、24 h、72 h、144 h处死SD大鼠,免疫组织化学染色法检测IL-23和IL-17蛋白在视网膜上的表达及分布情况;Western blot和ELISA检测分析各时间点视网膜IL-23和IL-17的表达水平和变化规律。结果免疫组织化学染色法显示IL-23和IL-17蛋白在正常对照组视网膜中表达极少,而在模型组表达明显增多。两者主要表达于视网膜神经节细胞层和内核层细胞的细胞浆。Western blot法检测发现IL-23和IL-17蛋白在RIRI视网膜中的表达明显增强,IL-23蛋白在建模后24 h表达最强,而IL-17蛋白在建模后72 h表达达到最高峰。ELISA法结果显示建模后12 h、24 h、72 h、144 h,模型组视网膜IL-23和IL-17蛋白的表达水平均较正常对照组明显升高,差异均有显著统计学意义(均为P<0.01)。结论 IL-23和IL-17在RIRI模型SD大鼠视网膜中表达明显增强,IL-23/IL-17通路作为致病因素,通过加重视网膜炎症反应参与RIRI的病理损伤。

Objective To observe the expression of interleukin-23 （IL-23） and in- terleukin-17 （IL-17）in rat retina after ischemia-reperfusion injury, and explore their role in retinal ischemia-reperfusion injury （RIRI）. Methods A total of 75 Sprague-Dawley rats were divided into normal control group （NCG） and model group（MG）. The rats in NCG group were not treated. The rats in MG group were induced the RIRI models with normal saline by anterior chamber perfusion. Immunotdstochemistry staining was used to detect specific lesions expression of IL-23 and IL-17 at 12 hours,24 hours,72 hours, 144 hours after RIRI. Both levels of IL-23 and IL-17 protein and changes were quantified by Western-blot and Enzyme-linked immunosorbent assay（ELISA）. Results Immuno- histochemistry staining results showed that IL-23 and IL-17 expressions were very few in NCG group, but were increased significantly in MG group. The expression of positive signal mainly located in ganglion cell layer and iimer nuclear layer of retina. Western- blot test analysis revealed that IL-23 and IL-17 expressions were increased significantly after RIRI,IL-23 was the highest at 24 hours,while IL-17 was the highest at 72 hours re- spectively after RIRI. ELISA assay showed that the expression of IL-23, IL-17 in MG group were increased at 12 hours,24 hours,72 hours, 144 hours after RIRI,and had a significant statistical difference compared to NCG group （all P 〈 0.01 ）. Coneluslon The expression levels of IL-23 and IL-17 are increased significantly in retina after RIRI, which maybe suggests that both aggravate inflammatory response and participate in RIRI,leading to retinal damage.