CAJ | 학술논문

目的观察单免疫球蛋白白细胞介素1受体相关蛋白(SIGIRR)对内毒素-脂多糖(LPS)诱导的人气道上皮细胞株H292炎症反应的影响.方法以人气道上皮细胞株H292为研究对象,实验分为两组:A组(实验组,SIGIRR过表达组)；B组(对照组,空载体转染组).将含有SIGIRR cDNA全长的真核表达载体瞬时转染H292细胞24h后,使SIGIRR在H292细胞中过表达,并设立空载体转染组作为对照组；分别采用逆转录-聚合酶链反应(RT-PCR)和Western blot法检测SIGIRR表达.于基因转染24h后给予LPS刺激H292细胞,分别于加入脂多糖(LPS)后3、6、12、24h各时间点,Western blot法检测转染前后核因子-κB(NF-κB)转录活性的改变,酶联免疫吸附试验(ELISA)检测转染前后炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β及IL-6表达水平的变化.结果转染SIGIRR表达载体的H292细胞,其SIGIRR表达量显著上升.LPS刺激后,Western blot法检测可见,对照组NF-κB转录活性明显升高；而实验组转染SIGIRR表达载体后NF-κB转录活性下降.ELISA实验结果表明,过表达SIGIRR的H292细胞在接受LPS刺激后其炎性因子TNF-α、IL-1β和IL-6蛋白表达水平明显低于对照组.结论 SIGIRR过表达可减轻H292细胞对LPS诱导的炎性反应,抑制H292细胞株产生TNF-α、IL-1β和IL-6等炎性因子.
목적 관찰단면역구단백백세포개소1수체상관단백(SIGIRR)대내독소-지다당(LPS)유도적인기도상피세포주H292염증반응적영향.방법 이인기도상피세포주H292위연구대상,실험분위량조:A조(실험조,SIGIRR과표체조)；B조(대조조,공재체전염조).장함유SIGIRR cDNA전장적진핵표체재체순시전염H292세포24h후,사SIGIRR재H292세포중과표체,병설립공재체전염조작위대조조；분별채용역전록-취합매련반응(RT-PCR)화Western blot법검측SIGIRR표체.우기인전염24h후급여LPS자격H292세포,분별우가입지다당(LPS)후3、6、12、24h각시간점,Western blot법검측전염전후핵인자-κB(NF-κB)전록활성적개변,매련면역흡부시험(ELISA)검측전염전후염성인자종류배사인자-α(TNF-α)、백세포개소(IL)-1β급IL-6표체수평적변화.결과 전염SIGIRR표체재체적H292세포,기SIGIRR표체량현저상승.LPS자격후,Western blot법검측가견,대조조NF-κB전록활성명현승고；이실험조전염SIGIRR표체재체후NF-κB전록활성하강.ELISA실험결과표명,과표체SIGIRR적H292세포재접수LPS자격후기염성인자TNF-α、IL-1β화IL-6단백표체수평명현저우대조조.결론 SIGIRR과표체가감경H292세포대LPS유도적염성반응,억제H292세포주산생TNF-α、IL-1β화IL-6등염성인자.
Objective To investigate the effect of single immunoglobulin interleukin-1 receptor related protein (SIGIRR)on lipopolysaccharide (LPS)-induced inflammatory reaction of human airway epithelial cell strain H292.Methods H292 cells were divided into two groups:group A (experimental group,SIGIRR over-expression group) ; group B (control group,empty vector transfection group).24 h after transient transfection of H292 cells with eukaryotic expression vectors containing full length of SIGIRR cDNA,SIGIRR was overexpressed in H292 cells.The SIGIRR expression was detected by using reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting,respectively.24 h after transfection,H292 cells were stimulated by LPS.Western blotting and enzyme linked immunosorbent assay (ELISA) were done in two groups before transfection and 3,6,12,24 h after LPS stimulation.Western blotting was used to detect the change of transcription activity of nuclear factor-κB (NF-κB) before and after the transfection,and enzyme linked immunosorbent assay (ELISA) was used to detect the changes in the expression levels of tumor necrosis factor-α (TNF-α),interleukin (IL)-1β and IL-6.Results The SIGIRR expression was significantly increased in SIGIRR cDNA-transfected H292 cells.After the stimulation with LPS,Western blotting revealed that NF-κB transcription activity was elevated in controlgroup,while decreased in experimental group.ELISA results showed that after LPS stimulation,TNF-α,IL-1β and IL-6 levels in H292 cells were significantly lower in the experimental group than in the control group.Conclusion Over-expression of SIGIRR may reduce the inflammatory reaction of H292 cells induced by LPS and inhibit the expression of TNF-α,IL-1β and IL-6 in H292 cells.