白介素-17A在急性百草枯中毒小鼠肺损伤中的作用  

The role of interleukin-17A in acute paraquat induced lung injury in mice

作　　者：宋宏伟[1] 杨琛[1] 刘晓伟[1] 刘志[1] 

机构地区：[1]中国医科大学附属第一医院急诊科,沈阳110001

出　　处：《中华急诊医学杂志》2017年第3期278-284,共7页Chinese Journal of Emergency Medicine

基　　金：辽宁省科学技术计划项目（2013225303）

摘　　要：目的探讨白介素-17A（IL-17A）在急性百草枯中毒致小鼠肺损伤中的作用。方法120只健康SPF级ICR雄性小鼠，随机（随机数字表法）分为3组：生理盐水对照组（NS）、百草枯染毒组（PQ）和抗体中和组（PQ＋Ah），每组40只。PQ组和PQ＋Ab组小鼠以25mg／kg的剂量一次性经口灌胃5mg／mL的PQ溶液建立中毒模型，抗体中和组每只小鼠于染毒后立即经腹腔注射5μg IL-17A中和性抗体；NS组小鼠给予相应量的生理盐水。染毒后每组按8h、1d、3d、5d、7d各取6只存活小鼠进行实验，测定小鼠肺湿干比（W／D），HE染色光镜下观察小鼠肺组织病理变化并评分，酶联免疫吸附法（ELISA）检测血清细胞因子IL-17A、白介素-22（IL-22）、白介素-6（IL-6）、转化生长因子-β（TGF-B）含量，免疫组化法检测肺组织白介素-23受体（IL-23R）表达，实时荧光定量PCR（qRT—PCR）法测定肺组织维甲酸相关孤核受体（RORyt）mRNA表达，SPSS22．0软件分析数据。结果PQ灌胃后，PQ组和PQ＋Ab组小鼠肺W／D显著升高（P〈0．01），肺组织出现急性损伤的病理改变，血清细胞因子（IL-17A、IL-22、IL-6、TGF—B）含量增多（P〈0．05）、肺组织中IL-23R和RORYt mRNA的表达升高（P〈0．01）；抗体中和IL-17A后，PQ＋Ab组小鼠肺W／D降低（P〈0．05），肺病理损伤减轻，血清细胞因子水平下降（P〈0．05），肺组织中IL-23R和RORYt mRNA的表达降低（P〈0．05）。结论IL-17A参与急性百草枯中毒致小鼠肺损伤过程，阻断IL-17A能够明显减轻小鼠肺急性损伤。Objective To investigate the role of interleukin-17A （IL-17A） in acute paraqnat （PQ） -induced lung injury in mice. Methods A total of 120 healthy SPF grade ICR male mice were randomly （ random number） divided into three groups （ n = 40 in each ） ： normal saline control group （NS）, PQ poisoning group （PQ） and antibody neutralization group （PQ ＋ Ab）. Mice of PQ group and PQ ＋ Ab group were given 5 mg/mL PQ by one garage in a dosage of 25 mg/kg body weight, and 5μg IL-17A neutralizing antibody intra-peritoneally administered immediately after PQ poisoning in PQ ＋ Ab group; Equivalent volume of normal saline instead of PQ was given to mice of NS group. Six survival mice from each group were taken for experiment at 8 h, 1 d, 3 d, 5 d, 7 d after PQ poisoning： Wet to dry ratio （W/D） of lung was determined in mice of each group. HE staining of lung tissue was used to observe the histopathological changes under the light microscope and the pathological scores were graded; Serum interleukin-17A （IL-17A）, interleukin-22 （IL-22）, interleukin-6 （IL-6）, transforming growth factor-β （TGF-13） were detected with enzyme linked immunosorbent assay （ELISA）; Expression of interleukin-23receptor （IL-23R） in lung tissue was determined with immunohistochemical; real-time fluorescence quantification PCR （qRT-PCR） was used to detect the expression of retinoic acid related solitary nuclear receptors＇ mRNA in lung tissue. Results After administration of PQ, W/D ratio increased （ P 〈 0. 01 ）, lung injury was observed in mice of PQ and PQ ＋ Ab groups, levels of cytokines （IL-17A, IL-22, IL-6 and TGF-β） in serum elevated （P 〈 0. 05）, and the expressions of IL-23R mRNA and RORYt mRNA increased （P 〈0. 01）. But in PQ ＋ Ab group, W/D ratio decreased （P 〈 O. 05）, lung injury was alleviated, the levels of cytokines （IL-17A, IL-22, IL-6 and TGF-β） decreased （P 〈 0. 05 ）, and the expressions of IL- 23R mRNA and RORyt mRNA reduced （P

关 键 词：百草枯 白介素-17A 小鼠 急性肺损伤 TH17细胞 

分 类 号：R563[医药卫生—呼吸系统] R595.4[医药卫生—内科学]