Effect of TRPV1 channel on proliferation and apoptosis of airway smooth muscle cells of rats

Abstract

Airway remodeling is an important pathological feature of asthma and the basis of severe asthma. Proliferation of airway smooth muscle cells (ASMCs) is a major contributor to airway remodeling. As an important Ca²⁺ channel, transient receptor potential vanilloid 1 (TRPV1) plays the key role in the cell pathological and physiological processes. This study investigated the expression and activity of TRPV1 channel, and further clarified the effect of TRPV1 channel on the ASMCs proliferation and apoptosis in order to provide the scientific basis to treat asthmatic airway remodeling in clinical practice. Immunofluorescence staining and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of TRPV1 in rat ASMCs. Intracellular Ca²⁺ was detected using the single cell confocal fluorescence microscopy measurement loaded with Fluo-4/AM. The cell cycles were observed by flow cytometry. MTT assay and Hoechst 33258 staining were used to detect the proliferation and apoptosis of ASMCs in rats respectively. The data showed that: (1) TRPV1 channel was present in rat ASMCs. (2) TRPV1 channel agonist, capsaicin, increased the Ca²⁺ influx in a concentration-dependent manner (EC₅₀=284.3±58 nmol/L). TRPV1 channel antagonist, capsazepine, inhibited Ca²⁺ influx in rat ASMCs. (3) Capsaicin significantly increased the percentage of S+G₂M ASMCs and the absorbance of MTT assay. Capsazepine had the opposite effect. (4) Capsaicin significantly inhibited the apoptosis, whereas capsazepine had the opposite effect. These results suggest that TRPV1 is present and mediates Ca²⁺ influx in rat ASMCs. TRPV1 activity stimulates proliferation of ASMCs in rats.

Keywords

transient receptor potential vanilloid 1 / airway smooth muscle cells / intracellular calcium / proliferation / apoptosis

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