焦炉逸散物致支气管上皮细胞 Th17细胞因子改变

摘要

OBJECTIVE To find the infla mmation bio markers induced by coke oven e missions (COE),we investigated the changes of T helper 17 (Th17 )cytokines in hu man bronchial epithelial (16HBE)cells.METHODS 16HBE cells were exposed to organic extracts of COE collected fro m co-king plant at the concentrations of 5,10 and 20 mg·L -1 for 24 h or 5 d to establish short-term and long-term cell models,respectively.Cell viability was measured by MTT assay and infla mmatory da mage was assessed by lactate dehydrogenase assay (LDH).The cytokines in culture supernatant sa mples was detected by co mmercial hu man Th17 cytokine panel kit.RESULTS COE Can induce infla mmation in COE 20 mg·L -1 group and no expression on IL-17 F and IL-1 β.The concentration of IL-10 was 1 .25 ± 0.54,1 .39 ±0.13 and (1 .90 ±0.73)pg·mL -1 in COE 5,10 and 20 mg·L -1 group showing good con-centration-effect relationship (r=0.98,P <0.05 ).IL-23 expression was found only higher at 10 and 20 mg·L -1 and the concentrations were 3.38 ±3.90 and (1 .74 ±2.00 )pg·mL -1 ,respectively.In 16HBE cells treated by COE for 5 d,elevated expression of IL-17A was found in COE 5 and 10 mg·L -1 group,and there was statistically sigificant difference between COE 10 mg·L -1 and DMSO group (P<0.05).Elevated concentration of IL-17F of 10.2 ±1 1 .78 and (6.79 ±7.84)pg·mL -1 was found in COE 5 and 10 mg·L -1 group.The concentration of IL-10 was 1 .71 ±0.02,1 .49 ±0.25 and (2.82 ± 0.33)pg·mL -1 in COE 5,10 and 20 mg·L -1 group,respectively.We found increased IL-1 βexpression with concentration of 2.72 ±0.62,2.25 ±0.33 and (0.93 ±0.21 )pg·mL -1 in COE 5,10 and 20 mg·L -1 group with negative dose-response relationship.We also found more elevated TNF-αlevels in the 5 d than in the 24 h model with no COE specific relationship.CONCLUSION COE induces expression changes of Th17 cytokines profile in 16HBE cells,including IL-23 and IL-1 βfor early and long-term infla mmation,respectively.IL-10 may be a candidate marker for population study on COE induced infla mmatory injury.%目的：检测焦炉逸散物（COE）致 Th17细胞因子谱表达改变，探讨其致炎性损伤过程中的生物标志物。方法焦化厂炉顶采集 COE，COE 5，10和20 mg·L -1处理人支气管上皮细胞（16HBE），分别为24 h 和连续染毒5 d。MTT 方法检测细胞存活率，乳酸脱氢酶释放实验检测细胞炎性损伤，高通量悬浮芯片测定细胞上清中 Th17细胞相关多种因子的改变情况。结果染毒24 h 模型，COE 20 mg·L -1组出现明显的炎性损伤改变，未检测到 IL-17F 和 IL-1β的表达；IL-10在 COE 5，10和20 mg·L -1组的表达量依次为1．25±0．54，1．39±0．13和（1．90±0．73）pg·mL -1，呈较好的剂量-效应关系（r ＝0．98，P ＜0．05）；IL-23只在 COE 10和20 mg·L -1组表达，表达量依次为3．38±3．90和（1．74±2．00）pg·mL -1，其他细胞因子未发现明显剂量依赖性改变。染毒5 d 模型中，IL-17A 在 COE 5和10 mg·L -1组的表达量升高，且在 COE 10 mg·L -1组有统计学意义（P＜0．05）。IL-17F 仅在 COE 5和10 mg·L -1组表达，分别为10．2±11．78和（6．79±7．84）pg·mL -1；IL-10在 COE 5，10和20 mg·L -1组的表达量依次为1．71±0．02，1．49±0．25和（2．82±0．33）pg·mL -1；COE 5，10和20 mg·L -1组 IL-1β表达量分别为2．72±0．62，2．25±0．33和（0．93±0．21）pg·mL -1，与处理剂量负相关；COE 染毒组 TNF-α表达量均高于染毒24 h 组，但无剂量相关性。结论COE 可以影响16HBE 细胞中 Th17细胞因子的表达改变，其中 IL-23和 IL-1β可以分别反映COE 早期和长期作用的炎性损伤；IL-10可以作为 COE 致炎性损伤生物标志物。