摘要
目的 利用 c DNA芯片技术分析内毒素活化的小鼠腹腔巨噬细胞早期和晚期基因表达 ,以更全面了解内毒素在感染、创伤反应中通过巨噬细胞介导的炎症和免疫反应。方法 以未刺激的和用 1mg/ L脂多糖 ( L PS)分别刺激 2 h(早期 )和 2 4 h(晚期 )的小鼠腹腔巨噬细胞制备 33P标记的 c DNA探针 ,并分别与小鼠 c DNA表达芯片 (含 1176个已知基因 )杂交。结果 巨噬细胞活化早期的 3倍差异表达基因为 6 9个 ,其中4 4个上调 ,2 5个下调 ;巨噬细胞活化晚期的 3倍差异表达基因中有 11个上调 ,2 6个下调 ;只有 8个基因同时出现于活化早期和晚期的差异表达基因中。许多转录因子、细胞内信号转导调节蛋白、炎症细胞因子和细胞凋亡相关基因的表达均发生了明显的调节变化。发现 BTB和 CNC同源 1( BACH1)、早期生长反应蛋白 2( EGR2 )、E4 7反应蛋白 1( EIP1)、Ngfi A结合蛋白 2 ( NAB2 )、成髓细胞白血病癌基因样蛋白 ( MYBL2 )、神经纤维癌蛋白基因 1( NF1)、睫状神经营养因子 ( CNTF)和 Sema4 A等一些以前未曾报道与 L PS诱导的巨噬细胞活化相关的基因。结论 采用 c DNA芯片技术了解内毒素诱导的巨噬细胞活化早期和晚期的综合基因表达信息 ,有助于更好地了解感染。
Objective To study the early and late changes in mRNA expression in macrophages in response to lipopolysaccharide (LPS) with a cDNA microarray approach using the Clontech Atlas microarray . Methods mRNA was isolated from unstimulated control and LPS stimulated murine peritoneal macrophages at 2 hours and 24 hours poststimulation,converted to 33 P radiolabeled cDNA, and hybridized to mouse array membranes. Results In macrophages being stimulated for 2 hours, 69 out of 1 176 genes were found to differ by over 3fold compared with the control. Among them 44 genes were upregulated and 25 genes were downregulated. In macrophages stimulated for 24 hours, 11 genes were upregulated and 26 genes were downregulated compared with the control. Only 8 genes were identified both at 2 hours and at 24 hours poststimulation. The expressions of many genes encoding transcription factor, cytokines,cell signaling modulators and apoptosis associated proteins were found to have changed. Some genes that were not previously linked to this model, such as bricabrac(BTB) and capncollar(CNC) homology 1(BACH1) , early growth response protein 2(EGR2), E47 interaction protein 1(EIP1), NgfiA binding protein 2 (NAB2), myeloblastosis oncogenelike protein(MYBL2), neurofibromatosis 1(NF1), ciliarry neurotropic factor(CNTF) and semaphorin 4A(Sema4A). Conclusion This study has allowed us to identify genes that may potentially be regulated by LPS at early and late phase in macrophages. These may contribute to better understanding of the mechanism underlying LPS or bacteria induced inflammatory and immune response following infection and trauma.
出处
《中国危重病急救医学》
CAS
CSCD
2004年第6期338-344,共7页
Chinese Critical Care Medicine
基金
国家自然科学基金资助项目 ( 3 0 10 0 176)
