Increased TMEM16A Involved in Alveolar Fluid Clearance After Lipopolysaccharide Stimulation

Abstract

We detected the expression of TMEM16A in LA795 cells and mouse lung tissue by RT-PCR, Western blot, and RNA interference techniques. TMEM16A expression was significantly increased by LPS stimulation in LA795 cells and in mouse lung tissue. Moreover, the LPS-induced TMEM16A expression enhancement in lung tissue was much more prominent in the alveolar epithelial region than in bigger airway epithelial cells. The typical TMEM16A current was recorded, and LPS treatment significantly enhances the current amplitude in LA795 cells. TMEM16A shRNA or TMEM16A inhibitor (T16Ainh-A01) did not affect alveolar fluid clearance (AFC), while co-application of T16Ainh-A01 induced a stronger AFC inhibition than LPS alone. LPS notably and synchronously enhanced Akt phosphorylation (p-Akt) and TMEM16A expression in a time-dependent manner in LA795 cells. Taken together, our results suggest that TMEM16A maybe plays an important role in pathological conditions of LPS-induced ALI as a protective protein.

Inhibition of transmembrane member 16A calcium‐activated chloride channels by natural flavonoids contributes to flavonoid anticancer effects

“…Confocal images were taken with Olympus Fluoview Fvl000 Microsystems. TMEM16A fluorescence intensity was analysed by the ImageJ system (Li et al, ).…”

Calcium-Activated Chloride Channel ANO1/TMEM16A: Regulation of Expression and Signaling

“…ANO1 expression was increased by angiotensin II in human umbilical vein endothelial cells (HUVECs) ( Ma et al, 2017 ); however, angiotensin II inhibited ANO1 expression in vascular smooth muscle cells ( Zhang et al, 2015 ), demonstrating cell-specific effects. Among the pathological stimuli, ANO1 expression was transiently induced by the bacterial toxin lipopolysaccharide (LPS) in human lung adenocarcinoma A549 cells ( Zhang et al, 2014 ), in cultured mouse lung cancer cell line LA795, and in mouse alveolar epithelial cells in vivo after intraperitoneal injection of LPS ( Li H. et al, 2016 ). Bacterial pyocyanin and supernatants from Pseudomonas aeruginosa culture stimulated ANO1 expression in human bronchial epithelial cells ( Caci et al, 2015 ).…”

“…Likewise, brain-derived neurotrophic factor (BDNF), acting through tropomyosin-related kinase (Trk) receptors ( Binder and Scharfman, 2004 ), induced robust Cl – currents, and these effects were blocked by pharmacological inhibition of ANO1 or by ANO1 knockout ( Hong et al, 2019 ). Interestingly, LPS increased currents in whole-cell patch recordings in cultured mouse lung cancer cell line LA795, and this was reduced by ANO1 knockdown; however, the role of calcium in these currents was not demonstrated ( Li H. et al, 2016 ).…”

Calcium-activated chloride channels: structure, properties, role in physiological and pathological processes

“…Основными физиологическими эндогенными факторами, вызывающими увеличение экспрессии TMEM16А, являются интерлейкины, эпидермальный фактор роста (epidermal growth factor (EGF) и ангиотензин II [115]. Бактериальный токсин липополисахарид вызывает увеличение экспрессии TMEM16А при развитии рака лёгкого у человека и в альвеолярных эпителиальных клетках мышей [116,117].…”