胆红素对早产儿NOD样受体2介导的炎症信号通路的影响  

Effect of bilirubin on inflammatory signaling pathway mediated by NOD - like receptor 2 in premature infants

作　　者：叶丹妮[1] 管欣娴[1] 高沙沙[1] 杨艳[1] 俞生林[1] Ye Danni;Guan Xinxian;Gao Shasha;Yang Yan;Yu Shenglin(Department of Neonatology,Children＇s Hospital of Soochow University,Suzhou 215025,Jiangsu Province,China)

机构地区：[1]苏州大学附属儿童医院新生儿科,215025

出　　处：《中华实用儿科临床杂志》2018年第14期1079-1082,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：江苏省妇幼健康科研项目（F201710）

摘　　要：目的探讨胆红素对早产儿NOD样受体2（NOD2）介导的炎症信号通路的影响。方法收集2016年4月至2017年1月苏州大学附属儿童医院新生儿科收治的15例早产儿外周血各2 mL，分离外周血单个核细胞，平均分为6组，分别为空白对照组（A组）、胞壁酰二肽（MDP）对照组（B组）、102 μmol/L胆红素组（C组）、102 μmol/L胆红素＋MDP组（D组）、153 μmol/L胆红素＋MDP组（E组）、255 μmol/L胆红素＋MDP组（F组）。A组、B组加入缓冲液，C组、D组、E组、F组分别加入102 μmol/L、102 μmol/L、153 μmol/L、255 μmol/L胆红素溶液孵育1 h，弃上清，A组、C组加入培养基液，B组、D组、E组、F组加入等量MDP激动剂刺激细胞24 h，分别收集细胞和上清液，实时定量-聚合酶链反应测定细胞中NOD2 mRNA的表达量，酶联免疫吸附法测上清液中白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）的表达水平。结果经MDP激动剂和102 μmol/L、153 μmol/L、255 μmol/L的胆红素刺激后，NOD2 mRNA的表达量（7.16±3.08、6.19±1.99、7.02±4.04、6.84±1.81）与空白对照组（7.46±3.70）比较差异均无统计学意义（均P〉0.05）。经MDP激动剂刺激后，B组IL-6表达水平（5.13±2.36）较空白对照组（3.84±1.44）明显上升，差异有统计学意义（P〈0.05）；2组TNF-α水平比较差异无统计学意义（4.85±2.47比4.04±2.26，P〉0.05）。经102 μmol/L的胆红素刺激后，IL-6、TNF-α表达水平与空白对照组比较差异均无统计学意义（3.26±1.06比3.84±1.44、3.45±1.84比4.04±2.26，均P〉0.05）；经153 μmol/L、255 μmol/L胆红素刺激后，IL-6、TNF-α表达水平（IL-6：2.58±1.33、2.16±0.94；TNF-α：2.32±1.49、2.42±1.42）较空白对照组（3.84±1.44、4.04±2.26）明显降低，差异均有统计学意义（均P〈0.05）。结论胆红素对NOD2无明显抑制作用，低浓度胆红素对IL-6、TNF-α无明显抑制作用，但高浓度胆红素ObjectiveTo investigate the effect of bilirubin on inflammatory signaling pathway mediated by NOD-like receptor 2（NOD2） in premature infants.MethodsFifteen cases of premature infants hospitalized at the Department of Neonatology, Children′s Hospital of Soochow University from April 2016 to January 2017, were selected, and 2 mL peripheral blood were collected from 15 cases of premature infants, and the mononuclear cells were isolated and divided into 6 groups, including blank control group （group A）, muramyl dipeptide（MDP） group （group B）, 102 μmol/L bilirubin group（group C）, 102 μmol/L bilirubin＋ MDP group （group D）, 153 μmol/L bilirubin＋ MDP group （group E）, 255 μmol/L bilirubin＋ MDP group （group F）. Group A and group B were stimulated by buffer, group C, group D, group E and group F were stimulated by 102 μmol/L, 102 μmol/L, 153 μmol/L, 255 μmol/L bilirubin, respectively.The supernatant was discarded after 1 h, then the medium was added to group A and group C, and the rest of the 4 groups were agonisted with MDP, the cells were stimulated for 24 h, and then the cells and supernatant fluids were collected respectively, the expression levels of NOD2 mRNA in the cells were determinated by real time-PCR, and the expression levels interleukin-6（IL-6） and tumor necrosis factor-α（TNF-α） in the supernatant was determinated by enzyme linked immunosorbent assay.ResultsThe expression levels of NOD2 mRNA had no obvious changes after being stimulated by MDP or by different concentrations of bilirubin（7.16±3.08, 6.19±1.99, 7.02±4.04, 6.84±1.81） compared to those of the blank control group（7.46±3.70）（all P〉0.05）. After being stimulated by MDP, the expression level of IL-6 （5.13±2.36）was significantly higher than that of the blank control group（3.84±1.44）, and the difference was statistically significant（P〈0.05）, but TNF-α had no obvious changes（4.85±2.47 vs.4.04±2.26, P〉0.05）. The expression levels of IL-6 and TNF-α had no o

关 键 词：胆红素 NOD样受体2 炎症信号通路 婴儿 早产 

分 类 号：R722.6[医药卫生—儿科]