光气吸入性肺损伤大鼠细胞因子群的变化及乌司他丁的干预

Dynamic changes of a group of cytokines in phosgene-induced lung injury and the function of ulinastatin

目的 建立大鼠光气吸入性肺损伤观察模型,观察大鼠血浆细胞因子群的动态变化及不同剂量乌司他丁干预的作用.方法 对104只雄性SD大鼠进行随机分组为空气对照组,乌司他丁干预对照组,光气染毒组及不同剂量乌司他丁干预组,每组8只.动态恒量染毒,干预组染毒后即刻经腹腔注射药物,并在2、6、24 h后处死实验动物,取肺组织行病理观察;检测肺湿/干比;半定量反转录聚合酶链式反应（RT-PCR）检测mRNA表达,取血浆行Bio-Plex细胞因子检测共18种.结果 与空气对照组比较,光气染毒早期（2 h）大鼠血浆中白细胞介素（IL）-1α、IL-6粒-巨噬细胞集落刺激因子（GM-CSF）、肿瘤坏死因子（TNF-α）、γ-干扰素（INF-γ）,巨噬细胞炎症蛋白3α（MIP-3α）,血管内皮生长因子（VEGF）浓度明显升高,随时间推移逐步下降,差异有统计学意义（P＜0.05）.血浆IL-4、IL-10浓度早期（2、6h）下降,后期（24 h）上升至正常或高于正常水平,血浆IL-13含量早期（2 h）下降不明显,后期（24 h）仍有上升,差异均有统计学意义（P＜0.05）.乌司他丁干预组的IL-1α、IL-6、GM-CSF、INF-γ、MIP-3α、VEGF、ING-α水平在不同时间点有不同程度下降,IL-4、IL-10、IL-13血浆浓度上升,差异有统计学意义（P＜0.05）.乌司他丁干预组肺损伤程度较染毒组有不同程度改善,且高剂量干预组改善更明显.乌司他丁干预组的肺组织IL-10,IL-4,IL-13-mRNA相对表达量上调与血浆细胞因子结果一致.结论 大鼠光气吸入性肺损伤时部分炎症细胞因子群随时间推移产生动态变化.乌司他丁可改善大鼠光气吸入性肺损伤的程度,调节炎症因子合成及释放,抑制炎症反应,且呈剂量依赖性.

Objective To investigate the dynamic changes of a group of cytokines in phosgene-induced lung injury and the function of different dose of ulinastatin through animal experiment.Methods 104 male SD rats were randomly assigned into the control group,ulinastatin control group,phosgene treatment groups and different dose of ulinastatin intervention groups,8 rats each group.Treatment groups were dynamic constant exposure in phosgene,and immediately injected ulinastatin intraperitoneal,and then the experimental animal,the lung tissue biopsy,lung wet/dry ratio,RT-PCR detection,the plasma for detection of Bio-Plex 18 cytokines.Results Compared with the control group,plasma concentrations of IL-1α,IL-6,GM-CSF,TNF-α,INF-γ,MIP-3α,VEGF were increased significantly first （2 h）,and gradually decreased with the passage of time,the difference was statistically significant （P＜0.05）.Plasma concentrations of IL-4,IL-10 were decreased earlier（2 h,6 h） and increased later （24 h） （P＜ 0.05）.The change of plasma concentration of IL-13 was not obvious earlier （2 h） and still rising later （24h）,the difference was statistically significant （P＜0.05）.After drug intervention,the levels of pro-inflammatory cytokines declined and the levels of anti-inflammatory cytokines raise by different degrees at different times in ulinastatin intervention groups,the difference was statistically significant.The degree of lung injury was improved than the phosgene treatment groups and better in high dose of ulinastatin intervention group.The expression of IL-10,IL-4,IL-13-mRNA of tissue increased in accordance with plasma results.Conclusion A group of cytokines are dynamicly changed in phosgene-induced lung injury by time.High dose of ulinastatin can improved phosgene-induced lung injury,regulate the synthesis and release of inflammatory cytokines and inhibit inflammatory react in a dose-dependent manner.