摘要
目的:研究哮喘发作时外周血嗜酸细胞(EOS)与细胞粘附及免疫调节相关基因的差异表达。方法:利用Percoll密度梯度离心分离哮喘患者发作时与治疗缓解外周血嗜酸性粒细胞,提取总RNA,并利用SuperSMARTcDNASynthesis技术合成cDNA第1链及优化扩增第2链。按照SSH常规方法构建cDNA消减文库,筛选阳性克隆并加以鉴定以得到带有差异表达的基因克隆。结果:成功构建具有高消减效率的哮喘发作患者外周血EOScDNA文库,筛选鉴定后确认15个差异表达基因,测序并同源性比对分别为夏科-莱登结晶蛋白(CLCprotein,galectin-10)、假定mRNA前剪接调节子female-lethal(2D)[putativepre-mRNAsplicingregulatorfemale-lethal(2D)]、水通道蛋白9(AQP-9)、IL-8、slingshot磷酸酶(SSH-2L)、蛋白磷酸酶1催化亚基β亚型(PP1CB)、RNA解旋酶HELZ、β2-微球蛋白(β2-MG)及一个线粒体相关基因。结论:这些基因的差异表达证明了EOS的趋化、迁移、粘附及免疫调节功能参与了哮喘病理生理调节,对这些途径的干预可能为未来哮喘靶向性治疗提供依据。
AIM： To determine differentially expressed genes associated with cell adhesion and immune regulation in peripheral blood eosinophils from asthmatic patients. METHODS： Peripheral blood eosinophils were isolated from the asthmatic patients at the time of exacerbation and after improvement. Total RNA was＇extracted. Super SMART PCR cDNA was synthesized, suppression subtractive hybridization （SSH） and PCR - select differential screening technology were used to detect expressed genes. The differentially expressed genes were sequenced. RESULTS： High efficiency subtractive cDNA library was constructed successfully. Differential screening identified 15 differentially expressed genes, which were Charcot- Leyden crystal protein （CLC protein; galectin - 10）, putative pre - mRNA splicing regulator female - lethal （2D）, aquaporin 9 （AQP9）, IL-8, slingshot 2L （SSH-2L）, PP1 catalytic subunit, beta isoform, helicase with zinc finger domain （ HELZ）, β2 - microglobin （ β2 - MG） and a gene associated with mitochondrion. CONCLUSION： increased expression of these genes might be associated with eosinophil migration, adhesion and immune regulation. Intervention of these pathways may provide a theoretical.base for future new targeting treatment for asthma.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2006年第7期1382-1387,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金项目资助(No.30270593)
广东省自然科学基金项目资助(No.04105757)
关键词
哮喘
嗜酸细胞
免疫调节
抑制性消减杂交
Asthma
Eosinophils
Immune regulation
Suppression subtractive hybridization
