Ikari Jun, Nelson Amy J, Obaid Jannah, Giron-Martinez Alvaro, Ikari Kumiko, Makino Fumihiko, Iwasawa Shunichiro, Gunji Yoko, Farid Maha, Wang Xingqi, Basma Hesham, Demeo Dawn, Feghali-Bostwick Carol, Holz Olaf, Rabe Klaus, Liu Xiangde, Rennard Stephen I
Pulmonary, Critical Care, Sleep and Allergy Medicine, University of Nebraska Medical Center, Omaha, Nebraska, United States of America.
Respirology, Graduate School of Medicine, Chiba University, Chiba, Japan.
PLoS One. 2017 Sep 7;12(9):e0184039. doi: 10.1371/journal.pone.0184039. eCollection 2017.
Alterations in microRNA (miRNA) expression may contribute to COPD pathogenesis. In COPD, lung fibroblast repair functions are altered in multiple ways, including extracellular mediator release. Our prior study revealed miR-503 expression is decreased in COPD lung fibroblasts, although the exact role played by miR-503 is undetermined. The current study examined a role of miR-503 in cytokine, growth factor and fibronectin production by lung fibroblasts from patients with and without COPD. Primary adult lung fibroblasts were isolated from patients with or without COPD. MiR-503 expression and interleukin (IL)-6, -8, PGE2, HGF, KGF, VEGF and fibronectin release were examined with or without inflammatory cytokines, IL-1β and tumor necrosis factor (TNF)-α. MiR-503 expression was decreased in COPD lung fibroblasts. The expression of miR-503 was positively correlated with %FVC, %FEV1, and %DLco as well as IL-6, -8, PGE2, HGF, KGF, and VEGF in the absence or presence of IL-1ß/TNF-α. In addition, IL-8 and VEGF release from COPD lung fibroblasts were increased compared to those from control. Exogenous miR-503 inhibited VEGF release from primary adult and fetal lung fibroblasts but not IL-8 release. As expected, COPD fibroblasts proliferated more slowly than control fibroblasts. MiR-503 did not affect proliferation of either control or COPD lung fibroblasts. MiR-503 inhibition of VEGF protein production and mRNA was mediated by direct binding to the 3' untranslated region of VEGF mRNA. Endogenous miR-503 was differently regulated by exogenous stimulants associated with COPD pathogenesis, including IL-1ß/TNF-α, TGF-ß1 and PGE2. Endogenous miR-503 inhibition augmented VEGF release by IL-1ß/TNF-α and TGF-ß1 but not by PGE2, demonstrating selectivity of miR-503 regulation of VEGF. In conclusions, reduced miR-503 augments VEGF release from lung fibroblasts from patients with COPD. Since VEGF contributes to disturbed vasculature in COPD, altered miR-503 production might play a role in modulating fibroblast-mediated vascular homeostasis in COPD.
微小RNA(miRNA)表达的改变可能促成慢性阻塞性肺疾病(COPD)的发病机制。在COPD中,肺成纤维细胞的修复功能以多种方式改变,包括细胞外介质的释放。我们之前的研究表明,COPD肺成纤维细胞中miR-503的表达降低,尽管miR-503的确切作用尚不确定。当前的研究探讨了miR-503在有或无COPD患者的肺成纤维细胞产生细胞因子、生长因子和纤连蛋白中的作用。从有或无COPD的患者中分离出原代成人肺成纤维细胞。在有或无炎性细胞因子白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α的情况下,检测miR-503的表达以及IL-6、-8、前列腺素E2(PGE2)、肝细胞生长因子(HGF)、角质形成细胞生长因子(KGF)、血管内皮生长因子(VEGF)和纤连蛋白的释放。COPD肺成纤维细胞中miR-503的表达降低。在有或无IL-1β/TNF-α的情况下,miR-503的表达与用力肺活量(%FVC)、1秒用力呼气容积(%FEV1)、一氧化碳弥散量(%DLco)以及IL-6、-8、PGE2、HGF、KGF和VEGF呈正相关。此外,与对照组相比,COPD肺成纤维细胞释放的IL-8和VEGF增加。外源性miR-503抑制原代成人及胎儿肺成纤维细胞释放VEGF,但不抑制IL-8的释放。正如预期的那样,COPD成纤维细胞的增殖比对照成纤维细胞更慢。miR-503对对照或COPD肺成纤维细胞的增殖均无影响。miR-503对VEGF蛋白产生和mRNA的抑制作用是通过直接结合VEGF mRNA的3'非翻译区介导的。内源性miR-503受到与COPD发病机制相关的外源性刺激物的不同调节,包括IL-1β/TNF-α、转化生长因子(TGF)-β1和PGE2。内源性miR-503的抑制增强了IL-1β/TNF-α和TGF-β1诱导的VEGF释放,但不增强PGE2诱导的VEGF释放,这表明miR-503对VEGF的调节具有选择性。总之,miR-503减少会增加COPD患者肺成纤维细胞释放VEGF。由于VEGF导致COPD中血管紊乱改变,miR-503产生的改变可能在调节COPD中成纤维细胞介导的血管稳态中起作用。