nasal mucosa hnepc  (PromoCell)

Journal: Clinical and Experimental Otorhinolaryngology

Article Title: Ghrelin Downregulates Lipopolysaccharide/ Leptin-Induced MUC5AC Expression in Human Nasal Epithelial Cells

doi: 10.21053/ceo.2022.00857

Figure Lengend Snippet: Effect of ghrelin on lipopolysaccharide (LPS)-induced MUC5AC expression in human nasal epithelial cells. (A, B) The results of reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR show that ghrelin significantly induced GHSR1a messenger RNA (mRNA) expression. (C) The results of RT-PCR and real-time PCR show that ghrelin significantly inhibited LPS-induced MUC5AC mRNA expression. LPS-induced MUC5AC expression started to be maximally suppressed at ta ghrelin concentration of 0.1 μM. (D) The results of real-time PCR show that the maximal inhibition of LPS-induced MUC5AC mRNA expression by ghrelin (0.1 μM) occurred after 2 hours. (E, F) The results of enzyme-linked immunosorbent assay also show that ghrelin significantly inhibited LPS-induced MUC5AC protein production. The images are representative of three separate experiments performed in triplicate. The bars indicate the mean±standard deviation of three independent experiments performed in triplicate. GHSR1a, growth hormone secretagogue receptor 1a; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. a) P <0.05 compared to the baseline value. b) P <0.05 is compared with samples treated with LPS (1 μg/mL) alone.

Article Snippet: Informed consent from patients was waived due to this study is conducted using human nasal mucosa epithelial cells commercially available from PromoCell (C-12620, donor age/sex/race 14/female/Caucasian, nasal mucosa, freezing medium Cryo-SFM).

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Concentration Assay, Inhibition, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Clinical and Experimental Otorhinolaryngology

Article Title: Ghrelin Downregulates Lipopolysaccharide/ Leptin-Induced MUC5AC Expression in Human Nasal Epithelial Cells

doi: 10.21053/ceo.2022.00857

Figure Lengend Snippet: The regulatory mechanism of ghrelin on lipopolysaccharide (LPS)-induced MUC5AC expression in human nasal epithelial cells. (A, B) The results of reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR show that ghrelin inhibited LPS-induced MUC5AC expression, and this inhibition was abolished by D-Lys-3-growth hormone-releasing peptide 6 (D-Lys-3-GHRP-6). (C, D) The results of enzyme-linked immunosorbent assay and immunofluorescence staining also show that ghrelin significantly inhibited LPS-induced MUC5AC protein production, while the inhibitory effect of LPS-induced MUC5AC protein production was abolished by D-Lys-3-GHRP-6. (E) Western blot results show that ghrelin significantly inhibited LPS-activated extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinases (MAPKs). These ghrelin-mediated changes in MAPK activation were also abolished by D-Lys-3-GHRP-6. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; DAPI, 4´,6-diamidino-2-phenylindole; p-ERK1/2, phosphorylated ERK 1/2; p-p38, phosphorylated p38. a) P <0.05 compared to the baseline value. b) P <0.05 compared with samples treated with LPS (1 μg/mL) alone. c) P <0.05 compared to samples treated with LPS (1 μg/mL) and ghrelin (0.1 μM).

Article Snippet: Informed consent from patients was waived due to this study is conducted using human nasal mucosa epithelial cells commercially available from PromoCell (C-12620, donor age/sex/race 14/female/Caucasian, nasal mucosa, freezing medium Cryo-SFM).

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Inhibition, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Western Blot, Activation Assay

Journal: Clinical and Experimental Otorhinolaryngology

Article Title: Ghrelin Downregulates Lipopolysaccharide/ Leptin-Induced MUC5AC Expression in Human Nasal Epithelial Cells

doi: 10.21053/ceo.2022.00857

Figure Lengend Snippet: Effect of ghrelin on leptin-induced MUC5AC expression in human nasal epithelial cells. (A) The results of reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR show that ghrelin significantly inhibited leptin-induced MUC5AC mRNA expression. Leptin-induced MUC5AC expression started to be maximally suppressed at a ghrelin concentration of 0.1 μM. (B) Real-time PCR results show that the maximal inhibition of leptin-induced MUC5AC mRNA expression by ghrelin (0.1 μM) occurred after 2 hours. (C, D) The results of enzyme-linked immunosorbent assay also showed that ghrelin significantly inhibited leptin-induced MUC5AC protein production. The images are representative of three separate experiments performed in triplicate. The bars indicate the mean±standard deviation of three independent experiments performed in triplicate. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. a) P <0.05 compared to the baseline value. b) P <0.05 compared with samples treated with lipopolysaccharide (LPS; 1 μg/mL) alone.

Article Snippet: Informed consent from patients was waived due to this study is conducted using human nasal mucosa epithelial cells commercially available from PromoCell (C-12620, donor age/sex/race 14/female/Caucasian, nasal mucosa, freezing medium Cryo-SFM).

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Concentration Assay, Inhibition, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Clinical and Experimental Otorhinolaryngology

Article Title: Ghrelin Downregulates Lipopolysaccharide/ Leptin-Induced MUC5AC Expression in Human Nasal Epithelial Cells

doi: 10.21053/ceo.2022.00857

Figure Lengend Snippet: The regulatory mechanism of ghrelin in leptin-induced MUC5AC in human nasal epithelial cells. (A, B) The results of reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR show that ghrelin inhibited leptin-induced MUC5AC expression, and this inhibition was abolished by D-Lys-3-growth hormone-releasing peptide 6 (D-Lys-3-GHRP-6). (C, D) The results of enzyme-linked immunosorbent assay and immunofluorescence staining also show that ghrelin significantly inhibited leptin-induced MUC5AC protein production, while the inhibitory effect of leptin-induced MUC5AC protein production was abolished by D-Lys-3-GHRP-6. (E) Western blot results show that ghrelin significantly inhibited leptin-activated extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinases (MAPKs). These ghrelin-mediated changes in MAPK activation were also abolished by D-Lys-3-GHRP-6. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; DAPI, 4´,6-diamidino-2-phenylindole; p-ERK1/2, phosphorylated ERK 1/2; p-p38, phosphorylated p38. a) P <0.05 compared to the baseline value. b) P <0.05 compared to samples treated with leptin (0.1 μg/mL) alone. c) P <0.05 compared to samples treated with leptin (0.1 μg/mL) and ghrelin (0.1 μM).

Article Snippet: Informed consent from patients was waived due to this study is conducted using human nasal mucosa epithelial cells commercially available from PromoCell (C-12620, donor age/sex/race 14/female/Caucasian, nasal mucosa, freezing medium Cryo-SFM).

Techniques: Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Expressing, Inhibition, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Western Blot, Activation Assay

Journal: Balkan Medical Journal

Article Title: Silencing SOX11 Alleviates Allergic Rhinitis by Inhibiting Epithelial-Derived Cytokines

doi: 10.4274/balkanmedj.galenos.2022.2022-9-31

Figure Lengend Snippet: STAT6 enhanced the transcription of SOX11 gene. (a) Schematic model of predicted binding sites of STAT6 on SOX11 promoter. (b) Effects of STAT6 on the transcriptional activity of truncated SOX11 promoter fragments. (c) SOX11 expression in AS1517499-treated HNEpCs measured by qRT-PCR. (d) SOX11 protein levels in AS1517499-treated HNEpCs obtained by Western blotting analysis. ** P < 0.01 vs. SOX11 promoter (−1757 to +20) + Vector-OE group; ## P < 0.01 vs. SOX11 promoter (−1757 to +20) + STAT6-OE group; §§ P < 0.01 vs. SOX11 promoter (−1334 to +20) + STAT6-OE group.

Article Snippet: The human nasal mucosa epithelial cell line HNEpC was procured from iCell Bioscience (Shanghai, China).

Techniques: Binding Assay, Activity Assay, Expressing, Quantitative RT-PCR, Western Blot, Plasmid Preparation

Journal: Balkan Medical Journal

Article Title: Silencing SOX11 Alleviates Allergic Rhinitis by Inhibiting Epithelial-Derived Cytokines

doi: 10.4274/balkanmedj.galenos.2022.2022-9-31

Figure Lengend Snippet: SOX11 and STAT6 expression were upregulated in IL-13-treated HNEpCs. (a) Schematic diagram of the culture and treatment of HNEpCs. (b) SOX11 expression measured by qRT-PCR. (c) SOX11 protein expression determined by Western blotting. (d) Immunofluorescence staining for SOX11 expression. (e) Representative images and relative protein levels of p-STAT6 and STAT6 obtained by Western blotting analysis.

Article Snippet: The human nasal mucosa epithelial cell line HNEpC was procured from iCell Bioscience (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining

Journal: Balkan Medical Journal

Article Title: Silencing SOX11 Alleviates Allergic Rhinitis by Inhibiting Epithelial-Derived Cytokines

doi: 10.4274/balkanmedj.galenos.2022.2022-9-31

Figure Lengend Snippet: Silencing SOX11 reduced the epithelial-derived cytokine expression and mucin level in IL-13-treated HNEpCs. (a) SOX11 expression in IL-13-induced HNEpCs measured by qRT-PCR. (b) SOX11 protein levels obtained by Western blotting analysis. (c-f) Expression of epithelial-derived cytokines measured by qRT-PCR. (g) MUC5AC levels in cell supernatants detected by ELISA.

Article Snippet: The human nasal mucosa epithelial cell line HNEpC was procured from iCell Bioscience (Shanghai, China).

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay