Lee Jong-Uk, Cheong Hyun Sub, Shim Eun-Young, Bae Da-Jeong, Chang Hun Soo, Uh Soo-Taek, Kim Young Hoon, Park Jong-Sook, Lee Bora, Shin Hyoung Doo, Park Choon-Sik
Department of Interdisciplinary Program in Biomedical Science Major, Soonchunhyang Graduate School, Bucheon, Korea.
Department of Genetic Epidemiology, SNP Genetics, Inc., Sogang University, Seoul, Korea.
Respir Res. 2017 Jan 5;18(1):3. doi: 10.1186/s12931-016-0493-6.
Idiopathic pulmonary fibrosis (IPF) is characterized by the complex interaction of cells involved in chronic inflammation and fibrosis. Global gene expression of a homogenous cell population will identify novel candidate genes.
Gene expression of fibroblasts derived from lung tissues (8 IPF and 4 controls) was profiled, and ontology and functional pathway were analyzed in the genes exhibiting >2 absolute fold changes with p-values < 0.05. CCL8 mRNA and protein levels were quantified using real-time PCR and ELISA. CCL8 localization was evaluated by immunofluorescence staining.
One hundred seventy eight genes differentially expressed and 15 genes exhibited >10-fold change. Among them, 13 were novel in relation with IPF. CCL8 expression was 22.8-fold higher in IPF fibroblasts. The levels of CCL8 mRNA and protein were 3 and 9-fold higher in 14 IPF fibroblasts than those in 10 control fibroblasts by real-time PCR and ELISA (p = 0.022 and p = 0.026, respectively). The CCL8 concentrations in BAL fluid was significantly higher in 86 patients with IPF than those in 41 controls, and other interstitial lung diseases including non-specific interstitial pneumonia (n = 22), hypersensitivity pneumonitis (n = 20) and sarcoidosis (n = 19) (p < 0.005, respectively). Cut-off values of 2.29 pg/mL and 0.43 pg/mL possessed 80.2 and 70.7% accuracy for the discrimination of IPF from NC and the other lung diseases, respectively. IPF subjects with CCL8 levels >28.61 pg/mL showed shorter survival compared to those with lower levels (p = 0.012). CCL8 was expressed by α-SMA-positive cells in the interstitium of IPF.
Transcriptome analysis identified several novel IPF-related genes. Among them, CCL8 is a candidate molecule for the differential diagnosis and prediction of survival.
特发性肺纤维化（IPF）的特征是参与慢性炎症和纤维化的细胞之间存在复杂的相互作用。对同质细胞群体进行全基因组表达分析将有助于识别新的候选基因。
对来源于肺组织的成纤维细胞（8例IPF患者和4例对照）的基因表达进行分析，并对表达变化绝对值>2且P值<0.05的基因进行本体论和功能通路分析。使用实时PCR和ELISA对CCL8 mRNA和蛋白水平进行定量。通过免疫荧光染色评估CCL8的定位。
178个基因差异表达，15个基因表达变化>10倍。其中，13个基因是与IPF相关的新基因。IPF成纤维细胞中CCL8的表达高22.8倍。通过实时PCR和ELISA检测，14例IPF成纤维细胞中CCL8 mRNA和蛋白水平分别比10例对照成纤维细胞高3倍和9倍（P值分别为0.022和0.026）。86例IPF患者支气管肺泡灌洗液中CCL8浓度显著高于41例对照以及包括非特异性间质性肺炎（n = 22）、过敏性肺炎（n = 20）和结节病（n = 19）在内的其他间质性肺疾病患者（P值均<0.005）。CCL8浓度截断值为2.29 pg/mL和0.43 pg/mL时，区分IPF与正常对照及其他肺部疾病的准确率分别为80.2%和70.7%。CCL8水平>28.61 pg/mL的IPF患者生存期短于水平较低者（P = 0.012）。在IPF间质中，CCL8由α-SMA阳性细胞表达。
转录组分析鉴定出多个与IPF相关的新基因。其中，CCL8是用于鉴别诊断和生存预测的候选分子。
