纳米二氧化钛对IEC-6细胞炎症因子表达及JAK2/STAT3蛋白磷酸化的影响

doi:10.3969/j.issn.1004-616x.2019.02.006

癌变·畸变·突变 ›› 2019, Vol. 31 ›› Issue (2): 127-132.doi: 10.3969/j.issn.1004-616x.2019.02.006

纳米二氧化钛对IEC-6细胞炎症因子表达及JAK2/STAT3蛋白磷酸化的影响

刘静^1,2, 张小强^1,2, 王旭^1,2, 苗歆雨^1,2, 卢晓星^1,2

1. 东南大学公共卫生学院, 江苏南京 210009;
2. 东南大学环境医学工程教育部重点实验室, 江苏南京 210009

收稿日期:2018-10-29 修回日期:2019-01-14 出版日期:2019-03-30 发布日期:2019-03-30
通讯作者: 张小强,E-mail:zhangxq7843@126.com E-mail:zhangxq7843@126.com
作者简介:刘静,E-mail:220163015@seu.edu.cn
基金资助:
中央高校基本科研业务费专项资金；江苏省普通高校研究生科研创新计划资助项目（SJCX17_0071）

Titanium dioxide nanoparticles on expression of inflammatory factors and phosphorylation of JAK2/STAT3 in IEC-6 cells

LIU Jing^1,2, ZHANG Xiaoqiang^1,2, WANG Xu^1,2, MIAO Xinyu^1,2, LU Xiaoxing^1,2

1. School of Public Health, Southeast University, Nanjing 210009;
2. Key Laboratory of Environmental Medicine Engineering of Ministry of Education, Southeast University, Nanjing 210009, Jiangsu, China

Received:2018-10-29 Revised:2019-01-14 Online:2019-03-30 Published:2019-03-30

摘要/Abstract

摘要： 目的：探讨纳米二氧化钛（nano-TiO₂）对肠道上皮IEC-6细胞中炎症因子表达及Janus激酶2/信号传导及转录激活因子3（JAK2/STAT3）蛋白磷酸化的影响。方法：取对数生长期的IEC-6细胞，分别加入浓度为0.0（对照组）、5.0、10.0、15.0、20.0 μg/mL的nano-TiO₂培养24 h，采用噻唑蓝（MTT）比色法检测各组细胞存活率；酶联免疫吸附实验（ELISA）检测各组上清液中白介素-1β（IL-1β）、白介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）水平；Western blot法测定各组细胞内JAK2和_STAT3蛋白表达及其磷酸化（p-JAK2和p-STAT3）水平；Image J软件分析蛋白条带灰度值。结果：nano-TiO₂处理组IEC-6细胞增殖活性与对照组相比差异无统计学意义（P>0.05）。与对照组相比，在各个nano-TiO₂处理浓度下，IEC-6细胞IL-6和TNF-α分泌水平均显著升高（P < 0.05）；10.0、15.0、20.0 μg/mL nano-TiO₂浓度处理后，细胞分泌IL-1β显著增加（P < 0.05）。Western blot法检测结果显示，nano-TiO₂处理组IEC-6细胞p-JAK2和p-STAT3蛋白表达升高，且其蛋白磷酸化水平和炎症因子表达水平呈正相关关系（r_{JAK2，IL-1β}=0.561，P < 0.05；r_JAK2，IL-6=0.711，P < 0.01；r_{JAK2，TNF-α}=0.675，P < 0.01；r_{STAT3，IL-1β}=0.782，P < 0.01；r_STAT3，IL-6=0.532，P < 0.05；r_{STAT3，TNF-α}=0.668，P < 0.01）。与对照组相比，15.0、20.0 μg/mL nano-TiO₂处理组IEC-6细胞中p-JAK2/JAK2和p-STAT3/STAT3灰度值比值明显升高（P < 0.05）。结论：nano-TiO₂可诱导IEC-6细胞分泌炎症因子和促进JAK2/STAT3蛋白磷酸化。

关键词: 纳米二氧化钛, 炎症, JAK2/STAT3, IEC-6细胞

Abstract: OBJECTIVE:To investigate the effects of nano-TiO₂ on expression of inflammatory cytokines and phosphorylation of JAK2/STAT3 in IEC-6 cells.METHODS:IEC-6 cells in cultures were treated with different doses (0.0,5.0,10.0,15.0,20.0 μg/mL) of nano-TiO₂ for 24 h.MTT assay was used to detect cell viability.The levels of inflammatory cytokines (IL-1β,IL-6 and TNF-α) were determined by ELISA assay.Protein expression and phosphorylated levels of JAK2 and STAT3 were measured by Western blot and the gray values of protein bands were assessed by Image J software.RESULTS:Compared with the control group,viability of IEC-6 cells was barely affected by nano-TiO₂.From the treatments with all doses of nano-TiO₂,IL-6 and TNF-α were highly secreted (P < 0.05),and significant secretion of IL-1β with the exception from the lowest dose of TiO₂,(P < 0.05).Western bolt results indicate the levels of p-JAK2 and p-STAT3 were increased after exposure of nano-TiO₂ and had a positive relationship with expressions of secreted inflammatory factors (r_JAK2,IL-1β=0.561,P < 0.05;r_JAK2,IL-6=0.711,P < 0.01;r_JAK2,TNF-α=0.675,P < 0.01;r_STAT3,IL-1β=0.782,P < 0.01;r_STAT3,IL-6=0.532,P < 0.05;r_STAT3,TNF-α=0.668,P < 0.01).The gray values of phosphorylated JAK2 and STAT3 increased when IEC-6 cells were exposed to 15.0 and 20.0 μg/mL nano-TiO₂(P < 0.05).CONCLUSION:Nano-TiO₂ exposure caused significant perturbation in expression of inflammatory cytokines and phosphorylation of JAK2 and STAT3 in IEC-6 cells.

Key words: nano-TiO₂, inflammation, JAK2/STAT3, IEC-6 cell

中图分类号:

刘静, 张小强, 王旭, 苗歆雨, 卢晓星. 纳米二氧化钛对IEC-6细胞炎症因子表达及JAK2/STAT3蛋白磷酸化的影响[J]. 癌变·畸变·突变, 2019, 31(2): 127-132.

LIU Jing, ZHANG Xiaoqiang, WANG Xu, MIAO Xinyu, LU Xiaoxing. Titanium dioxide nanoparticles on expression of inflammatory factors and phosphorylation of JAK2/STAT3 in IEC-6 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(2): 127-132.

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纳米二氧化钛对IEC-6细胞炎症因子表达及JAK2/STAT3蛋白磷酸化的影响

Titanium dioxide nanoparticles on expression of inflammatory factors and phosphorylation of JAK2/STAT3 in IEC-6 cells

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