McCluskey Elizabeth S, Liu Nathan, Pandey Abhimaneu, Marchetti Nathaniel, Sajjan Umadevi
Temple University.
Res Sq. 2023 Jul 25:rs.3.rs-3185241. doi: 10.21203/rs.3.rs-3185241/v1.
Airway basal cells 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 goblet cell metaplasia and the expression of pro-inflammatory cytokines in COPD models. This study assessed whether quercetin improves epithelial regeneration from COPD airway basal cells.
COPD airway basal cells were treated with DMSO or 1 μM quercetin for three days. The cells were then cultured at air/liquid interface (ALI) for up to 4 weeks. Basal cells from healthy donors cultured at air/liquid interface 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. Microarray analysis was conducted on DMSO or quercetin-treated COPD basal cells to identify differentially regulated genes (DEG) and the enriched biological pathways. Bronchial brushings from COPD patients treated with either placebo or quercetin for 6 months were used to confirm the effects of quercetin on gene expression.
Compared to DMSO, quercetin-treated COPD basal cells showed an increase in TER and regenerated the airway epithelium with more ciliated cells, and less goblet cells and IL-8. Comparison of DMSO- and quercetin-treated COPD basal cell transcriptomic profiles indicated that quercetin upregulated genes associated with tissue and epithelial development and differentiation. COPD patients treated with quercetin, but not placebo showed significantly increased expression of two developmental genes HOXB2 and ELF3, which were also increased in quercetin-treated COPD basal cells. Bronchial brushings from active smokers showed significantly increased mRNA expression of TGF-β and IL-8, and it was reduced 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会再生异常的气道上皮，这与几种参与上皮修复的基因表达降低有关。槲皮素可减少COPD模型中的杯状细胞化生和促炎细胞因子的表达。本研究评估了槲皮素是否能改善COPD气道基底细胞的上皮再生。
将COPD气道基底细胞用二甲基亚砜（DMSO）或1μM槲皮素处理三天。然后将细胞在气液界面（ALI）培养长达4周。将在气液界面培养的健康供体的基底细胞用作对照。在ALI培养8天时测定细胞的极化情况。通过流式细胞术和酶联免疫吸附测定法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或槲皮素处理的COPD基底细胞进行微阵列分析，以鉴定差异调节基因（DEG）和富集的生物学途径。使用接受安慰剂或槲皮素治疗6个月的COPD患者的支气管刷检样本，以确认槲皮素对基因表达的影响。
与DMSO相比，用槲皮素处理的COPD基底细胞的跨上皮电阻（TER）增加，并且再生的气道上皮中有更多的纤毛细胞，杯状细胞和IL-8较少。对用DMSO和槲皮素处理的COPD基底细胞转录组图谱的比较表明，槲皮素上调了与组织和上皮发育及分化相关的基因。接受槲皮素治疗而非安慰剂治疗的COPD患者显示两个发育基因HOXB2和ELF3的表达显著增加，在用槲皮素处理的COPD基底细胞中这两个基因也增加。现吸烟者的支气管刷检样本显示转化生长因子-β（TGF-β）和IL-8的mRNA表达显著增加，而在槲皮素治疗后降低。
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。
