摘要
目的观察转录因子FOXO1(forkhead box O1)对胰岛β细胞炎症因子产生的影响,探讨转录因子FOXO1在糖尿病发病机制中的作用。方法运用脂质体转染方法,将pcDNA-FOXO1真核表达载体瞬时转染至NIT-1细胞,通过Western blot检测细胞中FOXO1的过表达情况;以LPS分别处理重组质粒(pcDNA-FOXO1)、空质粒(pcDNA3.1)转染以及未转染的NIT-1细胞24 h后,应用ELISA法检测3组细胞致炎因子IL-1β、TNF-α表达情况。以LPS和PBS分别处理NIT-1细胞24 h后,ELISA测定两组细胞致炎因子IL-1β、TNF-α表达情况,Western blot比较两组细胞FOXO1蛋白表达及磷酸化水平的差异。结果 FOXO1在瞬时转染48 h的NIT-1细胞中实现了过表达;经LPS刺激后,重组质粒转染的NIT-1细胞产生的IL-1β和TNF-α显著高于空质粒转染和未转染组(P<0.01);经LPS处理24 h后的NIT-1细胞产生了致炎因子IL-1β和TNF-α,而PBS处理组未见表达。LPS处理组中FOXO1蛋白表达高于PBS处理组,但磷酸化水平显著低于PBS处理组(P<0.01)。结论炎症状态下,过表达转录因子FOXO1可增加胰岛β细胞致炎因子的产生,这主要与非磷酸化FOXO1的相对增多有关,它有望成为糖尿病抗炎治疗的新靶点。
In this study,we aimed to investigate the influence of transcription factor forkhead box O1（FOXO1） on the production of proinflammatory cytokines from pancreatic β cells and to analyze its relationship with diabetes.Firstly,recombinant plasmid pcDNA-FOXO1 or pcDNA3.1 was transfected in NIT-1 cells.24 hours later,Western blot was used to analyze the expression of FOXO1 in transfectants or wildtype.Secondly,the proinflammatory cytokines IL-1β and TNF-α were measured in the supernatants of three groups,following treatment with LPS,respectively.Thirdly,we used Western blot to analyze the expression of FOXO1 and phosphorylation of FOXO1 in NIT-1 cells treated either by LPS or PBS.We found the FOXO1 protein overexpressed in pcDNA-FOXO1-transfected NIT-1 cells 48 hours after transfection.After 24 hours of LPS stimulation for,the protein levels of IL-1β and TNF-α were significantly increased in FOXO1 overexpressed NIT-1 as compared to blank transfectants or wild type（P〈0.01）.The expression levels of FOXO1 was elevated in LPS-treated NIT-1 cells,however the ratio of phosphorylation versus total FOXO1 levels was significantly reduced as compared to control（P〈0.01）.In conclusion,the overexpression of FOXO1 improves the production of proinflammatory cytokines of pancreatic β cells in inflammatory state,which is determined by phosphorylation of FOXO1,therefore FOXO1 may be a new target for anti-inflammation treatment of diabetes.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2011年第5期386-389,共4页
Immunological Journal
关键词
糖尿病
FOXO1
致炎因子
磷酸化
Diabetes
FOXO1
Proinflammatory cytokines
Phosphorylation
