中国临床医学2016，Vol.23Issue(6)：715-719,5.DOI:10.12025/j.issn.1008-6358.2016.20160822
过表达锌指蛋白A20可抑制肺泡巨噬细胞炎症反应
A20 regulates the inflammatory responses of alveolar macrophage
摘要
Abstract
Objective:To study the effects of A20 on the inflammatory response of alveolar macrophages and its regulation mechanism through establishing A 20 over‐expressing alveolar macrophage cell lines .Methods :Lentivirus‐mediated expression vector carrying A20 gene was constructed ,then it was transfected into rat alveolar macrophage cell lines (NR8383) , and the cell lines which stably overexpressed A20 gene were screened and cultured in vitro .Lipopolysaccharide (LPS ,1 μg/mL) was added into the medium to intervene ,the culture supernatants and cells were collected 0 .5 ,1 ,2 ,4 hours after stimulation ,ELISA method was used to determine the activity of cytokines (TNF‐α,IL‐1β) and nuclear factor (NF‐κB) . Western blotting method was used to detect A20 protein and nuclear p65 content ,and real‐time fluorescence quantitative PCR method was used to determine the content of A20 mRNA .Results:After LPS stimulation ,the levels of A20 protein and mRNA in A20 overexpression group (A20 group) and the normal control group (VEC group) both increased and reached the peak at 1 h ,and then gradually decreased .The A20 level of A20 group was significantly higher than that of VEC group (P<0 .05) .Compared with VEC group ,the levels of cytokines (TNF‐α,IL‐1β) in culture supernatants of A20 group significantly reduced (P<0 .05) ,and the DNA binding activity of NF‐κB and nuclear p65 content of A20 group also significantly reduced (P<0 .05) .Conclusions :A20 can inhibit the activity of NF‐κB and the secretion of TNF‐α and IL ‐1β,and then inhibit the inflammatory reaction activity of alveolar macrophages , thereby reducing the inflammatory response of acute respiratory distress syndrome (ARDS) .关键词
急性呼吸窘迫综合征/A20蛋白/肺泡/巨噬细胞/炎症反应Key words
acute respiratory distress syndrome/A20 protein/alveolar/macrophage/inflammatory response分类
医药卫生引用本文复制引用
朱晓丹,王颖,毕晶,童琳,刘洁,宋元林,白春学..过表达锌指蛋白A20可抑制肺泡巨噬细胞炎症反应[J].中国临床医学,2016,23(6):715-719,5.基金项目
国家自然科学基金青年基金（81300055，81500025），国家自然科学基金重点项目（30930090）．Supported by National Natural Science Youth Foundation of China （81300055，81500025） and Key Project of National Natural Science Foundation of China （30930090）． （）
