Effects of nitric oxide-mediated metabolic disruption on the proliferation of mouse airway progenitor cells

doi:10.11958/20222109

Abstract

Abstract:

Objective To evaluate the effect of nitric oxide (NO) on the proliferative function of airway club progenitor cells and underlying metabolic mechanisms. Methods The mouse airway progenitor club cells were sorted using flow sorting technique, then treated with 25 μmol/L DEA NONOate (diethylaminononanoate) for transcriptomic sequencing analysis. Mouse club cells were cultured using organoid culture technique. The cells were cultured in normal medium, 40 nmol/L simvastain (Hmgcr inhibitor), 0.2 μmol/L DON (Gmps inhibitor) or 1 mmol/L AOA (Gpt2 inhibitor) respectively. Microscopic photographs were taken with an inverted microscope on the day 8 of culture to observe the growth of club cell-derived organoids and to measure the organoid diameter and formation efficiency. Results DEA NONOate treatment of club cells resulted in significant changes in transcriptome expression (2 894 up-regulated genes and 3 270 down-regulated genes, fold ≥ 1.2), with significant decrease in expression levels of metabolism-related genes Hmgcr, Gmps and Gpt2 (P<0.01). Compared with the control group, the organoid diameter was significantly reduced (P<0.01), but the organoid forming efficiency was unchanged in the simvastain-treated group. Both organoid diameter and forming efficiency were significantly reduced in the DON-treated group (P<0.01) and the AOA-treated group (P<0.01). Conclusion NO inhibits the proliferative function of mouse airway club progenitor cells probably through its negative regulation of three metabolism-related pathways, Hmgcr, Gmps and Gpt2.

Key words: nitric oxide, cell proliferation, organoids, airway epithelial progenitor cells, cell metabolism

CLC Number:

R329.2+8，R562.2

Figures/Tables 6

Fig.1 Flow sorting strategy of mouse airway club cells and organoid culture model

Fig.2 The differential change genes of club cells after NO stimulation

Fig.3 The effect of NO on the expression of metabolic genes in mouse club cells

Fig.4 The effect of simvastain on the size of mouse club cell-derived organoids (×40)

Fig.5 The effect of DON on the size of mouse club cell-derived organoids (×40)

Fig.6 The effect of AOA on the size of mouse club cell-derived organoids (×40)

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