McCluskey Elizabeth S, Liu Nathan, Pandey Abhimaneu, Marchetti Nathaniel, Kelsen Steven G, Sajjan Umadevi S
Center for Inflammation and Lung Research, Lewis-Katz Medical School, Temple University, Philadelphia, PA, 19140, USA.
Department of Thoracic Medicine and Surgery, Temple University Health System, Philadelphia, PA, 19140, USA.
Respir Res. 2024 Mar 11;25(1):120. doi: 10.1186/s12931-024-02742-0.
Airway basal cells (BC) from patients with chronic obstructive pulmonary disease (COPD) regenerate abnormal airway epithelium and this was associated with reduced expression of several genes involved in epithelial repair. Quercetin reduces airway epithelial remodeling and inflammation in COPD models, therefore we examined whether quercetin promotes normal epithelial regeneration from COPD BC by altering gene expression.
COPD BC treated with DMSO or 1 µM quercetin for three days were cultured at air/liquid interface (ALI) for up to 4 weeks. BC from healthy donors cultured at ALI were used as controls. Polarization of cells was determined at 8 days of ALI. The cell types and IL-8 expression in differentiated cell cultures were quantified by flow cytometry and ELISA respectively. Microarray analysis was conducted on DMSO or 1 µM quercetin-treated COPD BC for 3 days to identify differentially regulated genes (DEG). Bronchial brushings obtained from COPD patients with similar age and disease status treated with either placebo (4 subjects) or 2000 mg/day quercetin (7 subjects) for 6 months were used to confirm the effects of quercetin on gene expression.
Compared to placebo-, quercetin-treated COPD BC showed significantly increased transepithelial resistance, more ciliated cells, fewer goblet cells, and lower IL-8. Quercetin upregulated genes associated with tissue and epithelial development and differentiation in COPD BC. COPD patients treated with quercetin, but not placebo showed increased expression of two developmental genes HOXB2 and ELF3, which were also increased in quercetin-treated COPD BC with FDR < 0.001. Active smokers showed increased mRNA expression of TGF-β (0.067) and IL-8 (22.0), which was reduced by 3.6 and 4.14 fold respectively after quercetin treatment.
These results indicate that quercetin may improve airway epithelial regeneration by increasing the expression of genes involved in epithelial development/differentiation in COPD.
This study was registered at ClinicalTrials.gov on 6-18-2019. The study number is NCT03989271.
慢性阻塞性肺疾病（COPD）患者的气道基底细胞（BC）可使异常气道上皮再生，这与几种参与上皮修复的基因表达降低有关。槲皮素可减轻COPD模型中的气道上皮重塑和炎症，因此我们研究了槲皮素是否通过改变基因表达促进COPD BC的正常上皮再生。
用二甲基亚砜（DMSO）或1 μM槲皮素处理COPD BC三天，然后在气液界面（ALI）培养长达4周。将在ALI培养的健康供体的BC用作对照。在ALI培养8天时测定细胞极化情况。分别通过流式细胞术和酶联免疫吸附测定法（ELISA）对分化细胞培养物中的细胞类型和白细胞介素-8（IL-8）表达进行定量分析。对用DMSO或1 μM槲皮素处理三天的COPD BC进行微阵列分析，以鉴定差异调节基因（DEG）。从年龄和疾病状态相似的COPD患者中获取支气管刷片，这些患者分别接受安慰剂（4例受试者）或2000 mg/天槲皮素（7例受试者）治疗6个月，以确认槲皮素对基因表达的影响。
与安慰剂相比，槲皮素处理的COPD BC显示跨上皮电阻显著增加，纤毛细胞增多，杯状细胞减少，IL-8降低。槲皮素上调了COPD BC中与组织和上皮发育及分化相关的基因。接受槲皮素治疗的COPD患者而非接受安慰剂治疗的患者，显示出两个发育基因HOXB2和ELF3的表达增加，在经槲皮素处理且错误发现率（FDR）<0.001的COPD BC中这两个基因也增加。现吸烟者显示转化生长因子-β（TGF-β）（0.067）和IL-8（22.0）的mRNA表达增加，槲皮素处理后分别降低了3.6倍和4.14倍。
这些结果表明，槲皮素可能通过增加COPD中参与上皮发育/分化的基因表达来改善气道上皮再生。
本研究于2019年6月18日在ClinicalTrials.gov注册。研究编号为NCT03989271。
