An inflammatory stimulus sensitizes TRPA1 channel to increase cytokine release in human lung fibroblasts

Yap, Jennifer Maries; Ueda, Takashi; Takeda, Norihisa; Fukumitsu, Kensuke; Fukuda, Satoshi; Uemura, Takashi; Tajiri, Tomoko; Ohkubo, Hirotsugu; Maeno, Ken; Ito, Yutaka; Kanemitsu, Yoshihiro

doi:10.1016/j.cyto.2020.155027

Cited by 25 publications

(24 citation statements)

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“…Quantitative RT-PCR of target genes were analyzed using 7900HT Fast real time PCR system (Applied biosystems Thermo Fisher Scientific, USA). We established specific primers as described previously (Additional file 1: Table S1) [19]. Relative gene expressions were calculated using 2 −ΔΔCt method after normalized with β-actin.…”

Section: Quantitative Rt-pcr Of Target Genesmentioning

confidence: 99%

“…Western blot analysis was performed as described previously [19], except for loaded protein volume (10 μg). Membranes were incubated with anti-GAPDH [4967S, Cell Signaling Technology (CST), Japan] or primary antibodies (Table 1) and followed by anti-rabbit IgG HRPconjugated antibody (7074S, CST) and/or anti-mouse IgG HRP-conjugated antibody (W402B, Promega, USA).…”

Section: Western Blot Analysismentioning

confidence: 99%

“…Transient receptor potential ankyrin 1 (TRPA1) is a major TRP channel in human lung fibroblasts. We have demonstrated that tumor necrosis factor-α (TNF-α) can regulate airway inflammation by modulating TRPA1 expression on lung fibroblasts [19]. TRPA1 is activated by an antioxidant substance, allyl isothiocyanate (AITC), along with a variety of risk factors for respiratory diseases, such as cigarette smoke extract, bacterial endotoxins and painful cold.…”

Section: Introductionmentioning

confidence: 99%

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AITC inhibits fibroblast-myofibroblast transition via TRPA1-independent MAPK and NRF2/HO-1 pathways and reverses corticosteroids insensitivity in human lung fibroblasts

et al. 2021

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Background Little is known on the role of transient receptor potential ankyrin 1 (TRPA1) in fibroblast—myofibroblast transition (FMT) that can lead to airway remodeling which is a major problem for severe asthma and fibrosis. Thus, this study investigated the effect of TRPA1 modulators on transforming growth factor beta 1(TGF-β1) -treated lung fibroblasts. Methods MRC-5 cells were preincubated with TGF-β1 for 24 h. TRPA1 agonist or antagonist were added and further incubated for 24 h. The changes in TRPA1 and alpha-smooth muscle actin (α-SMA) expressions by stimuli were evaluated using qRT-PCR, western blot and immunohistochemical analyses. Statistical significance was determined by using one- or two-way ANOVA, followed by Bonferroni’s post hoc analysis for comparison of multiple groups and paired 2-tailed Student’s t-test between 2 groups. Results MRC-5 cells treated by TGF-β1 significantly upregulated α-SMA mRNA expressions (P < 0.01), but downregulated TRPA1 gene expression (P < 0.001). Post-treatment of TRPA1 activator, allyl isothiocyanate (AITC), after TGF-β1 significantly downregulated the α-SMA gene induction (P < 0.01 at 24 h), protein expression (P < 0.05) and immunoreactivity with stress fibers (P < 0.05). On the other hand, TRPA1 antagonist HC-030031 did not prevent this effect, and instead tended to facilitate the suppressive effect of AITC when co-stimulated. AITC significantly increased phosphorylated- extracellular signal-regulated kinase (ERK) 1/2 and heme oxygenase (HO)-1 protein expressions (P < 0.05) in TGF-β1-treated cells. Combined inhibition with ERK1/2 mitogen-activated protein kinase (MAPK) and nuclear factor erythroid 2-related factor (NRF2) almost completely reversed AITC-induced α-SMA suppression (P < 0.05). Dexamethasone was not able to inhibit the upregulated α-SMA induction by TGF-β1. However, AITC improved dexamethasone-insensitive myodifferentiation in the presence of the corticosteroid (P < 0.01). Conclusion We found that AITC exerts protective effect on TGF-β1-induced α-SMA induction by activating ERK1/2 MAPK and NRF2/HO-1 pathways in lung fibroblasts. It also overcomes corticosteroids insensitivity in TGF-β1-induced α-SMA induction. TRPA1 antagonist modulates the suppressive effect, but not prevent it. AITC and TRPA1 antagonist may be therapeutic agents in treating chronic respiratory diseases.

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Section: Quantitative Rt-pcr Of Target Genesmentioning

confidence: 99%

Section: Western Blot Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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AITC inhibits fibroblast-myofibroblast transition via TRPA1-independent MAPK and NRF2/HO-1 pathways and reverses corticosteroids insensitivity in human lung fibroblasts

et al. 2021

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“…However, TRPA1-mediated effects via the sensory trigeminal afferentation of the upper airways also might contribute to our results in both models [70]. Besides the neuronal expression, TRPA1 is also located on epithelial, inflammatory and bronchial smooth muscle cells [6,47,48,50].…”

Section: Discussionmentioning

confidence: 74%

“…TRPA1 is also expressed on several non-neuronal cells [35], e.g., keratinocytes, macrophages and CD4+ lymphocytes [36][37][38], epidermal melanocytes, fibroblasts [39,40], urothelial and endothelial cells, as well as primary human osteoarthritic chondrocytes [41][42][43]. In the airways, TRPA1 is expressed on fibroblasts, tracheal, bronchial and alveolar epithelial cells, bronchial smooth muscle cells (SMC), as well as lymphocytes [17,[44][45][46][47][48][49][50].…”

Section: Introductionmentioning

confidence: 99%

Complex Regulatory Role of the TRPA1 Receptor in Acute and Chronic Airway Inflammation Mouse Models

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et al. 2020

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The Transient Receptor Potential Ankyrin 1 (TRPA1) cation channel expressed on capsaicin-sensitive afferents, immune and endothelial cells is activated by inflammatory mediators and exogenous irritants, e.g., endotoxins, nicotine, crotonaldehyde and acrolein. We investigated its involvement in acute and chronic pulmonary inflammation using Trpa1 gene-deleted (Trpa1−/−) mice. Acute pneumonitis was evoked by intranasal Escherichia coli endotoxin (lipopolysaccharide: LPS) administration, chronic bronchitis by daily cigarette smoke exposure (CSE) for 4 months. Frequency, peak inspiratory/expiratory flows, minute ventilation determined by unrestrained whole-body plethysmography were significantly greater, while tidal volume, inspiratory/expiratory/relaxation times were smaller in Trpa1−/− mice. LPS-induced bronchial hyperreactivity, myeloperoxidase activity, frequency-decrease were significantly greater in Trpa1−/− mice. CSE significantly decreased tidal volume, minute ventilation, peak inspiratory/expiratory flows in wildtypes, but not in Trpa1−/− mice. CSE remarkably increased the mean linear intercept (histopathology), as an emphysema indicator after 2 months in wildtypes, but only after 4 months in Trpa1−/− mice. Semiquantitative histopathological scores were not different between strains in either models. TRPA1 has a complex role in basal airway function regulation and inflammatory mechanisms. It protects against LPS-induced acute pneumonitis and hyperresponsiveness, but is required for CSE-evoked emphysema and respiratory deterioration. Further research is needed to determine TRPA1 as a potential pharmacological target in the lung.

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The regulation of TRPA1 expression and function by Th1 and Th2-type inflammation in human A549 lung epithelial cells

et al. 2023

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Background Transient Receptor Potential Ankyrin 1 (TRPA1) is a cation channel that mediates pain, itch, cough, and neurogenic inflammation in response to pungent compounds such as acrolein in cigarette smoke. TRPA1 is also activated by endogenous factors and promotes inflammation in asthma models. We have recently shown that TRPA1 is upregulated by inflammatory cytokines in A549 human lung epithelial cells. Here, we explored the effects of Th1 and Th2-type inflammation on TRPA1. Methods and results TRPA1 expression and function was studied in A549 human lung epithelial cells. To induce inflammation, the cells were exposed to a combination of cytokines TNF-α and IL-1β; and to model Th1 or Th2-type responses, IFN-γ or IL-4/IL-13 was added, respectively. TRPA1 expression (measured by RT-PCR and Western blot) and function (assessed by Fluo-3AM intracellular calcium measurement) was enhanced under the influence of TNF-α + IL-1β. IFN-γ further enhanced TRPA1 expression and function, whereas IL-4 and IL-13 suppressed them. The effects of IFN-γ and IL-4 on TRPA1 expression were reversed by the Janus kinase (JAK) inhibitors baricitinib and tofacitinib, and those of IL-4 also by the STAT6 inhibitor AS1517499. The glucocorticoid dexamethasone downregulated TRPA1 expression, whereas the PDE4 inhibitor rolipram had no effect. Under all conditions, TRPA1 blockade was found to reduce the production of LCN2 and CXCL6. Conclusions TRPA1 expression and function in lung epithelial cells was upregulated under inflammatory conditions. IFN-γ further increased TRPA1 expression while IL-4 and IL-13 suppressed that in a JAK-STAT6 dependent manner which is novel. TRPA1 also modulated the expression of genes relevant to innate immunity and lung disease. We propose that the paradigm of Th1 and Th2 inflammation is a major determinant of TRPA1 expression and function, which should be considered when targeting TRPA1 for pharmacotherapy in inflammatory (lung) disease.

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An inflammatory stimulus sensitizes TRPA1 channel to increase cytokine release in human lung fibroblasts

Cited by 25 publications

References 50 publications

AITC inhibits fibroblast-myofibroblast transition via TRPA1-independent MAPK and NRF2/HO-1 pathways and reverses corticosteroids insensitivity in human lung fibroblasts

AITC inhibits fibroblast-myofibroblast transition via TRPA1-independent MAPK and NRF2/HO-1 pathways and reverses corticosteroids insensitivity in human lung fibroblasts

Complex Regulatory Role of the TRPA1 Receptor in Acute and Chronic Airway Inflammation Mouse Models

The regulation of TRPA1 expression and function by Th1 and Th2-type inflammation in human A549 lung epithelial cells

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