Store-Operated Ca²⁺ Entry is involved in Transforming Growth Factor-β1 Facilitated Proliferation of Rat Airway Smooth Muscle Cells

Abstract

Objective. To investigate the role and underlying mechanisms of store-operated Ca²⁺ entry (SOCE) in mediating the promoting effect of transforming growth factor (TGF)-β1 on the proliferation of airway smooth muscle cells (ASMCs). Methods. Rat bronchial smooth muscle cells were cultured as we described previously. The intracellular Ca²⁺ concentration ([Ca²⁺]_i) of ASMCs was measured by laser confocal microscope Ca²⁺ fluorescence imaging with Fluo-3/AM. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and p27 expression assay were used to determine the proliferation rate of ASMCs. Results. We demonstrated that TGF-β1 (10 ng/ml) increased basal (Ca²⁺]_i) level, [Ca²⁺]_i rise induced by thapsigargin-induced Ca²⁺ release and SOCE in rat ASMCs. This effect of TGF-β1 on SOCE was not inhibited by glucocorticoid dexamethasone (DXM, 100 nM), antioxidant α-tocopherol (100 μM), and intermediate-conductance Ca²⁺-activated K⁺ channels (IK_Ca) inhibitor charybdotoxin (100 nM), suggesting that reactive oxygen species and IK_Ca channels might not mediate the effect of TGF-β1. TGF-β1 slightly increased the expression of Orai1 and STIM1, two important molecules involved in the molecule component and regulation of SOC channels, in the presence of 10% fetal bovine serum (FBS). The proliferation of ASMC stimulated with 2.5% FBS was promoted by TGF-β1, and partly inhibited by non-specific Ca²⁺ channel blocker SKF-96365 (10 μM) and Ni²⁺ (100 μM). DXM, α-tocopherol, and charybdotoxin had no effect on the proliferation promoted by TGF-β1. Conclusion. TGF-β1 promotes ASMC proliferation partly through increasing the expression and activity of SOC channels.

• airway smooth muscle cells
• proliferation
• store-operated Ca²⁺ entry
• transforming growth factor-β1

Acknowledgments

We thank B.Sc Bo-quan Pan (Biomedical Research Center, Wuhan University Medical College) for the technical support.