摘要
目的探讨p38 丝裂原激活蛋白激酶(MAPK)家族四种亚型对诱导型一氧化氮合酶(iNOS)基因的转录调控。方法以人胚胎肾293(HEK 293)细胞为靶细胞,采用脂质体(LPS)介导的细胞基因共转染技术、荧光素酶报告基因技术,分别将FLAG-tagged p38 MAPK 4种亚型、含有鼠iNOS基因启动子区的荧光素酶报告基因质粒(piNOS-Luc)、空载体(pcDNA3)、β-半乳糖苷酶表达质粒(pCMV-β)共转染,检测并比较荧光素酶相对活性。结果(1)未加刺激时,在HEK 293细胞中,p38 MAPK中仅有p38α能够诱导iNOS基因启动子的转录活性;(2)在LPS刺激下,p38 MAPK 4种亚型均能够诱导iNOS启动子的转录活性,其中,p38β所诱导的转录活性最高,p38α的诱导作用亦很明显。结论(1)LPS能够在HEK 293细胞中诱导iNOS基因转录活性;(2)在HEK 293细胞中,p38 MAPK参与了静息时及LPS刺激下对iN-OS基因的转录调控。
Objective To study the transcriptional regulation of inducible nitric oxide synthase (iNOS) gene by p38 mito-gen-activated protein kinase (MAPK). Method With human embryonic kidney (HEK) 293 cells as the target and the assis-tance of lipofectamine-mediated co-transfection techniques and luciferase reporter gene systems, FLAG-tagged p38 isoforms (namely FLAG-p38α, FLAG-p38β, FLAG-p38γand FLAG-p38δ) in pcDNA3, pcDNA3, piNOS-Luc and pCMV-βwere transfected into HEK 293 cells, and the relative activity of luciferase was subsequently tested. Results Highest luciferase ac-tivity occurred only in p38αgroup compared with the other three isoform groups under no stimulation. Under the stimulation by lipopolysaccharide (LPS), the luciferase activity of each group was obviously increased and the highest activity occurred in p38βgroup. Conclusion LPS can induce transcription and activation of iNOS gene, and p38 MAPK is involved in the tran-scription regulation of iNOS gene in HEK 293 cells.
出处
《第一军医大学学报》
CSCD
北大核心
2003年第3期206-209,共4页
Journal of First Military Medical University
基金
国家杰出青年科学基金(39925014)
国家自然科学基金(30030060
39800074)~~
