CAJ | 학술논문

目的研究人类白细胞抗原(HLA)-B27相关性前葡萄膜炎患者外周血单核细胞炎症通路差异基因的表达特征.方法实验研究.抽取3例HLA-B27阳性前葡萄膜炎患者及2例健康对照者外周血,分离后获得的单核细胞经含霍乱弧菌的脂多糖刺激后提取RNA,使用基因表达谱芯片进行检测,实时荧光定量PCR验证芯片结果,并通过基因功能分析、KEGG数据库等生物信息分析技术对筛选出的差异表达的基因进行分析.结果 HLA-B27阳性前葡萄膜炎患者的外周血单核细胞经脂多糖刺激后出现了1105个表达水平上调超过2倍的基因；而健康对照组只有25个.Geneontology聚类分析及信号转导通路分析结果显示,上调的基因主要参与蛋白转运、蛋白折叠,在炎症反应中起重要作用.脂多糖与Toll样受体4结合后激活了Toll样受体转导通路及霍乱弧菌感染相关通路,是位于整个炎症反应网络中较上游的转导通路.其中PIK3 CA、PIK3CB、AKT3及MAPK1基因可能是炎症反应中较关键的基因.结论 HLA-B27阳性前葡萄膜炎患者及健康对照者的外周血单核细胞经脂多糖刺激后差异表达的基因有显著的差异.Toll样受体转导通路在致病中起重要作用.
목적 연구인류백세포항원(HLA)-B27상관성전포도막염환자외주혈단핵세포염증통로차이기인적표체특정.방법 실험연구.추취3례HLA-B27양성전포도막염환자급2례건강대조자외주혈,분리후획득적단핵세포경함곽란호균적지다당자격후제취RNA,사용기인표체보심편진행검측,실시형광정량PCR험증심편결과,병통과기인공능분석、KEGG수거고등생물신식분석기술대사선출적차이표체적기인진행분석.결과 HLA-B27양성전포도막염환자적외주혈단핵세포경지다당자격후출현료1105개표체수평상조초과2배적기인；이건강대조조지유25개.Geneontology취류분석급신호전도통로분석결과현시,상조적기인주요삼여단백전운、단백절첩,재염증반응중기중요작용.지다당여Toll양수체4결합후격활료Toll양수체전도통로급곽란호균감염상관통로,시위우정개염증반응망락중교상유적전도통로.기중PIK3 CA、PIK3CB、AKT3급MAPK1기인가능시염증반응중교관건적기인.결론 HLA-B27양성전포도막염환자급건강대조자적외주혈단핵세포경지다당자격후차이표체적기인유현저적차이.Toll양수체전도통로재치병중기중요작용.
Objective To investigate the genes and signalling pathways located upstream of the inflammatory processes in human leukocyte antigen (HLA)-B27-associated acute anterior uveitis by gene expression microarray.Methods Experimental study.HLA-B27-positive and-negative monocytes isolated from human peripheral blood were stimulated with Vibrio cholera lipopolysaccharide (LPS).Gene expression microarrays were used to identify the differentially expressed genes.Differentially expressed (DE) genes were testified by real-time PCR and analyzed by a series of bioinformatics-based techniques such as Gene Ontology,Kyoto Encyclopedia of Genes and Genomes.Results Gene expression microarray analysis revealed marked differences between HLA-B27-positive acute anterior uveitis(AAU) and HLA-B27-negative healthy control peripheral monocytes in the genes that were upregulated in response to LPS stimulation with 1105 genes and 25 genes respectively.Gene Ontology enrichment and pathway analysis indicated that genes participating in protein transport and folding were essential to the inflammatory process.The LPS receptorToll-like receptor(TLR)4 induced TLR signalling pathway and pathway related to Vibrio cholerae infection were located upstream of the network and contribute to the overall response.Among the DE genes,PIK3 CA,PIK3CB,AKT3,and MAPK1 might play critical roles in inflammation.Conclusions Equivalent LPS stimulation induces a different response in HLA-B27-positive peripheral monocytes compared to normal control,suggesting that the TLR pathway is involved in the pathogenesis of HLA-B27-associated AAU.