Cordes Friederike, Brückner Markus, Lenz Philipp, Veltman Katharina, Glauben Rainer, Siegmund Britta, Hengst Karin, Schmidt Markus Alexander, Cichon Christoph, Bettenworth Dominik
*Department of Medicine B, University Hospital Münster, Münster, Germany; †Institute of Palliative Care, University Hospital Münster, Münster, Germany; ‡Center for Molecular Biology of Inflammation, Institute of Infectiology, University of Münster, Münster, Germany; and §Department of Medicine (Gastroenterology, Infectious Diseases, Rheumatology), Campus Benjamin Franklin, Charité-Universitätsmedizin Berlin, Berlin, Germany.
Inflamm Bowel Dis. 2016 Oct;22(10):2341-55. doi: 10.1097/MIB.0000000000000917.
Inflammatory bowel disease is a chronic-remittent disorder with the risk of disabling complications due to uncontrolled inflammation. Accurate biomarkers are needed to noninvasively monitor the disease course to tailor therapy. We evaluated the potential of the specific microRNA (miR)-320a to monitor disease activity in experimental colitis or patients with Crohn's disease and investigated its functional role in intestinal epithelial barrier formation.
The impact of miR-320a on intestinal barrier function was tested in vitro in T84 epithelial cells by transepithelial resistance measurement and quantitative real-time polymerase chain reaction analysis on inflammatory and microbial stimulation. Experimental colitis was studied in dextran sodium sulfate colitis, T-cell transfer colitis, and IL-10 mice. Disease course was monitored by body weight measurement, colonoscopy, and histological examination. MiR-320a expression during inflammation was assessed in T84 cells, murine blood, and colonic tissue and in peripheral blood from patients with Crohn's disease with active or quiescent disease.
MiR-320a transfection of T84 cells reinforced barrier integrity reflected by increased transepithelial resistance (P < 0.01) and inhibited barrier-destructive enteropathogenic Escherichia coli effects resulting in increased tight junction protein JAM-A expression (P = 0.02) and decrease of barrier integrity-destabilizing miR-320a target PPP2R5B (P < 0.001). Tumor necrosis factor-α and interleukin-1β stimulation increased a miR-320a epxression in T84 cells. MiR-320a level was increased in blood samples from colitic mice and patients with Crohn's disease showing a strong correlation with disease activity (r = 0.67).
MiR-320a strengthens intestinal barrier function in vitro and has the potential to monitor disease activity of colitic mice. Future studies are needed to further evaluate the potential of miR-320a in patients with inflammatory bowel disease.
炎症性肠病是一种慢性缓解性疾病，存在因炎症失控而导致致残性并发症的风险。需要准确的生物标志物来无创监测疾病进程以调整治疗方案。我们评估了特定微小RNA（miR）-320a在实验性结肠炎或克罗恩病患者中监测疾病活动的潜力，并研究了其在肠道上皮屏障形成中的功能作用。
通过跨上皮电阻测量以及对炎症和微生物刺激进行定量实时聚合酶链反应分析，在体外T84上皮细胞中测试miR-320a对肠道屏障功能的影响。在葡聚糖硫酸钠结肠炎、T细胞转移结肠炎和IL-10小鼠中研究实验性结肠炎。通过体重测量、结肠镜检查和组织学检查监测疾病进程。评估T84细胞、小鼠血液和结肠组织以及活动期或缓解期克罗恩病患者外周血中炎症期间的miR-320a表达。
T84细胞转染miR-320a可增强屏障完整性，表现为跨上皮电阻增加（P < 0.01），并抑制破坏屏障的肠致病性大肠杆菌的作用，导致紧密连接蛋白JAM-A表达增加（P = 0.02），以及降低破坏屏障完整性的miR-320a靶标PPP2R5B（P < 0.001）。肿瘤坏死因子-α和白细胞介素-1β刺激可增加T84细胞中miR-320a的表达。结肠炎小鼠和克罗恩病患者血液样本中的miR-320a水平升高，与疾病活动度呈强相关（r = 0.67）。
miR-320a在体外可增强肠道屏障功能，并有潜力监测结肠炎小鼠的疾病活动度。未来需要进一步研究评估miR-320a在炎症性肠病患者中的潜力。
