https://orcid.org/0000-0002-3609-784X
Figures & data
Figure 1. NEAT1 expression is elevated in nasal mucosal tissues from AR patients and positively correlated to IL-13 stimulation. (a) RT-qPCR showed NEAT1 expression levels in mucosal tissues from 30 patients with perennial AR and 30 patients with nonallergic rhinitis (NAR) were measured. (b) RT-qPCR showed NEAT1 expression in the AR-EXO and control-EXO. (c) Correlation of lncRNA NEAT1 expression with TNSS score. (d) RT-qPCR showed NEAT1 expression in human HNECs treated with 50 ng/mL IL-13 for 12, 24, or 48 h. *P < 0.05, **P < 0.01, ***P < 0.001
Figure 2. NEAT1 knockdown regulates IL-13-triggered inflammatory cytokine, mucus production, and apoptosis in HNECs. (a). RT-qPCR showed the level of NEAT1 in human HNECstransfected with si-NEAT1 or si-NC. (b–g) RT-qPCR and ELISA assay showed the expression levels of GM-CSF, eotaxin-1, and MUC5AC in IL-13-stimulated HNECs transfected with si-NEAT1 or si-NC. (h and i) CCK-8 and TUNEL assays indicated the cell viability and apoptosis in IL-13-treated HNECs transfected with si-NEAT1 or si-NC. *P < 0.05, **P < 0.01
Figure 3. Extracellular NEAT1 was transferred via incorporation in exosomes in HNECs. (a) The expression of NEAT1 was detected by RT-qPCR after cells were treated with RNase A or RNase A + 0.1% Triton X100 for 30 min. (b) The exosomes images secreted by IL-13-treated HNECs were showed by TEM scanning. (c) Size distribution of exosomes ranged from 30 to 120 nm. (d) The levels of exosomal marker proteins CD9 and CD63 were measured by Western blot in HNECs. (e) RT-qPCR analysis showed the expression of NEAT1 in exosomes extracted from IL-13-treated HNECs cells. **P < 0.01
Figure 4. NEAT1 silence attenuates the exosome-induced inflammatory response and apoptosis of HNECs. (a) RT-qPCR showed NEAT1 level in HNECs treated with PBS, exosome, exosome+si-NC, and exosome+si-NEAT1. (b–g) RT-qPCR and ELISA assay showed the mRNA expression levels of GM-CSF, eotaxin-1, and MUC5AC in HNECs treated with exosome, exosome+si-NC, and exosome+si-NEAT1. (h and i) CCK-8 and TUNEL assays indicated cell viability and apoptosis in different groups. *P < 0.05, **P < 0.01, ***P < 0.001
Figure 5. NEAT1 acts as a molecule sponge for miR-511. (a) The putative binding sites between NEAT1 and miR-511 were predicted by starBase website. (b) The luciferase activity of NEAT1-WT and NEAT1-Mut were measured in HNECs transfected with NC mimics, miR-511 mimics, NC inhibitor, or miR-511 inhibitor. (c) Correlation analysis between miR-511 and NEAT1 in nasal mucosal tissues from AR patients. (d) The expression of miR-511 was detected by RT-qPCR. (e) RT-qPCR showed miR-511 expression in HNECs transfected with si-NEAT1 or si-NC. **P < 0.01
Figure 6. miR-511 is a downstream regulator of NEAT1 in AR. (a) RT-qPCR showed miR-511 expression in HNECs transfected with si-NC, si-NEAT1, and si-NEAT1+miR-511 inhibitor. (b–g) RT-qPCR and ELISA assay showed the expression levels of GM-CSF, eotaxin-1, and MUC5AC in IL-13-treated HNECs transfected with si-NC, si-NEAT1, and si-NEAT1+ miR-511 inhibitor. (h and i) CCK-8 and TUNEL assays indicated the cell viability and apoptosis in IL-13-treated HNECs transfected with si-NC, si-NEAT1, and si-NEAT1+ miR-511 inhibitor. *P < 0.05, **P < 0.01
Figure 7. NR4A2 is a target of miR-511. (a) The putative binding sites between NR4A2 and miR-511 were predicted by TargetScan. (b) The luciferase activity of NR4A2-WT and NR4A2-Mut were measured in HNECs cells transfected with NC mimics or miR-511 mimics. (c) RT-qPCR showed NR4A2 expression in HNECs transfected with NC mimics, miR-511 mimics, NC inhibitor, or miR-511 inhibitor. (d) Correlation analysis between miR-511 and NR4A2 in nasal mucosal tissues from AR patients. **P < 0.01
Figure 8. NEAT1 regulates IL-13-induced dysfunction of HNECs via miR-511/NR4A2 axis. (a) NR4A2 expression was detected by RT-qPCR in HNECs transfected with NC inhibitor, miR-511 inhibitor, miR-511 inhibitor+si-NR4A2. (b–g) The levels of GM-CSF, eotaxin-1, and MUC5AC were determined by RT-qPCR and ELISA in IL-13-treated HNECs transfected with NC inhibitor, miR-511 inhibitor, miR-511 inhibitor + si-NR4A2. (h and i) TUNEL and CCK-8 assays indicated the cell apoptosis and viability in IL-13-treated HNECs transfected with NC inhibitor, miR-511 inhibitor, miR-511 inhibitor + si-NR4A2. (j) NR4A2 expression was measured by RT-qPCR in HNECs transfected with pcDNA3.1, pcDNA3.1-NEAT1, pcDNA3.1-NEAT1+ miR-511 mimics. (k) Correlation analysis between NEAT1 and NR4A2 in nasal mucosal tissues from AR patients. *P < 0.05, **P < 0.01
