细胞因子在内毒素诱导的前葡萄膜炎小鼠房水和血清中的表达

Expression of cytokine in aqueous humor and serum of mice with endotoxin induced anterior uveitis

目的通过分析内毒素诱导的C3H/HeN小鼠的前葡萄膜炎模型中房水和血清中肿瘤坏死因子-α(tumor necrosis fac-tor-α,TNF-α)、白细胞介素(interleukin,IL)-1、IL-6、IL-10、干扰素-γ(interferon-γ,IFN-γ)的浓度,探讨急性前葡萄膜炎可能的发病机制。方法取180只C3H/HeN小鼠,分为对照组(n=30)和实验组(n=150),将霍乱弧菌内毒素(18001株,古典生物型,小川血清型)溶于无菌生理盐水中,终浓度为2×103g.L-1,实验组小鼠足底注射100μL制作内毒素诱导的前葡萄膜炎模型;对照组注射等量生理盐水。于诱导后4h、8h、16h、24h、48h断颈处死实验组小鼠(各30只),以10只小鼠为一单样本,微量进样器抽取前房水;眼球摘除后微量进样器于眼窝处取外周血。对照组于注射生理盐水后即刻处死小鼠,样本收集方法同实验组。微球流式技术检测小鼠房水及血清中TNF-α、IL-1、IL-6、IL-10、IFN-γ的浓度。结果所有实验组小鼠均成功诱导出急性前葡萄膜炎。在小鼠房水中,TNF-α于4h达峰值,与对照组相比差异有统计学意义(P=0.002);4h、8h、16h及24h时,在房水和血清内的浓度差异均无统计学意义(均为P>0.05)。IL-1、IL-6在血清中均于16h达峰值,与对照组相比差异均有统计学(P=0.001、0.000);且诱导后各时间点在房水内的浓度均高于血清。IL-10在房水中于24h达峰值,与对照组相比差异有统计学意义(P=0.003),与相同时间点血清内的浓度相比差异亦有统计学意义(P=0.003)。IFN-γ于4h、24h在房水和血清中的表达差异有统计学意义(P=0.033、0.032)。各细胞因子在房水中的表达变化基本与炎症进程相符。结论细胞因子在眼内炎症发生过程中起着重要作用,网络调控将成为治疗方向。

Objective To investigate pathogenesis of acute anterior uveitis by analyzing the concentrations of tumor necrosis factor-α（TNF-α）,interleukin-1（IL-1）,IL-6,IL-10 and interferon-γ（IFN-γ）in aqueous humor and serum of C3H/HeN mice with endotoxin induced uveitis.Methods A total of 180 C3H/HeN mice were divided into control group（30 mice）and experimental group（150 mice）.Lipopolysaccharide（18001 strain,Classical biotype,Ogawa＇s serotype）was resolved into stroke-physiological saline solution,and the final concentration was 2×103 g·L-1.The mice of experimental group were injected at sole of foot with 100 μL lipopolysaccharide to induce uveitis models,and the mice of control group were injected with same amount of sodium chloride.Thirty mice of experimental group were sacrificed with breaking neck at 4 hours,8 hours,16 hours,24 hours and 48 hours after induction,respectively.Ten mice were treated as one sample,and aqueous humor was obtained by micro-injector.After removing eyeballs,peripheral blood was obtained by micro-injector.Mice in control group were immediately sacrificed after being injected with sodium chloride,and the way of collecting samples was same as experimental group.Cytometric bead array was used to detect the concentrations of TNF-α,IL-1,IL-6,IL-10 and IFN-γ in aqueous humor and peripheral blood of mice.Results All mice in experimental group were successfully induced with acute anterior uveitis.In aqueous humor of mice,TNF-α went to peak at 4 hours,and there was statistical difference with control group（P=0.002）;The concentration of TNF-α at 4 hours,8 hours,16 hours and 24 hours in aqueous humor had no statistical difference with those in serum（All P0.05）;IL-1 and IL-6 went to peak at 16 hours in serum,and there were statistical differences with control group（P=0.01,P=0.000）.Concentrations of IL-1 and IL-6 in aqueous humor were higher than those in serum;IL-10 in aqueous humor went to peak at 24 hours,which had significant differences with control group and with the concentration in serum at same time point（Both P=0.003）.The expression of IFN-γ at 4 hours and 24 hours in aqueous humor had significant difference with those in serum（P=0.033,P=0.032）.The expression of cytokines in aqueous humor was almost consistent with the process of inflammation.Conclusions Cytokines play an important role in the process of ocular inflammation,so the network control may be the therapeutic purpose.