Overexpression of miR-155-5p Inhibits the Proliferation and Migration of IL-13-Induced Human Bronchial Smooth Muscle Cells by Suppressing TGF-β-Activated Kinase 1/MAP3K7-Binding Protein 2

PURPOSE: Molecular mechanisms leading to asthma is still ill-defined. Though the function of microRNAs (miRNAs) in asthma was previously reported, the involvement of miR-155 in important features of this disease remains unknown. The present study was designed to uncover the probable involvement of miR-155-5p in the proliferation and migration of IL-13-induced human bronchial smooth muscle cells (BSMCs) and the intrinsic regulatory mechanism. METHODS: The effects of different concentrations of IL-13 on the proliferation and migration of BSMCs as well as the expression of miR-155-5p and its predicted target transforming growth factor (TGF)-β-activated kinase 1/MAP3K7-binding protein 2 (TAB2) were investigated. The effects of miR-155-5p on the proliferation and migration of interleukin (IL)-13-induced BSMCs was determined in vitro using BSMCs transfected with miR-155 mimic/inhibitor and induced by a high concentration of IL-13. The quantitative real-time polymerase chain reaction (qRTPCR) was employed for determining the expression of miR-155-5p and TAB2. Western blotting was applied to analyze the expression of TAB2 at the protein level. Cell proliferation and migration were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Transwell assays, respectively. RESULTS: The proliferation and migration of BSMCs were dose-dependently increased with IL-13 treatment. Contrariwise, IL-13 dose-dependently inhibited the expression of miR-155-5p in BSMCs. Mechanistic studies showed that inhibition of miR-155-5p further promoted the stimulatory effects of IL-13, whereas overexpression of miR-155 significantly inhibited these effects. In silico studies and luciferase reporter assays indicated that TAB2 was a negatively regulated miR-155-5p target. CONCLUSIONS: These results suggested that miR-155-5p-inhibit the IL-13-induced proliferation and migration of BSMCs by targeting TAB2 and that the IL-13/miR-155/TAB2 pathway could serve as a therapeutic target for pulmonary diseases, especially asthma.

YKL-40-induced IL-8 expression from bronchial epithelium leads to bronchial smooth muscle proliferation and migration / 中国免疫学杂志

Objective:To investigate YKL-40-mediated inflammation in human bronchial epithelial cells and analyzed the soluble factors secreted by bronchial epithelial cells exposed to YKL-40 that were responsible for increasing proliferation and migration of primary normal human bronchial smooth muscle cells (BSMCs).Methods:YKL-40-induced inflammation was assayed in two human bronchial epithelial cells (BEAS-2B cell line and primary human bronchial epithelial cells ,namely HBECs).In addition,we treated BEAS-2B cells and HBECs with YKL-40,and added the conditioned culture media ( YKL-40-BEAS-2B-CM) and ( YKL-40-HBECs-CM) to BSMCs.The proliferation and migration of BSMCs were determined by premixed WST-1 cell proliferation reagent and QCM chemotaxis migration assay ,respectively.Results: Bronchial epithelial cells treated with YKL-40 resulted in a significant increase of IL-8 production,but have no effect about RANTES ,Eotaxin and TNF-α.YKL-40-BEAS-2B-CM and YKL-40-HBECs-CM induced IL-8 was found to further stimulate proliferation and migration of BSMCs ,and the effects were inhibited after neutralizing IL-8.Conclusion:Through investigating the interaction of airway epithelium and smooth muscle ,our findings implicate that YKL-40 may be involved in the inflammation of asthma by induction of IL-8 from epithelium,subsequently contributing to BSMCs proliferation and migration.Moreover, inhibition of IL-8 signaling is a potential therapeutic target for YKL-40-induced inflammation and remodeling of asthma.

IL-13 R110Q, a Naturally Occurring IL-13 Polymorphism, Confers Enhanced Functional Activity in Cultured Human Bronchial Smooth Muscle Cells

PURPOSE: Interleukin (IL)-13, a Th2-type cytokine, plays a pivotal role in the pathogenesis of asthma through its direct effects on airway smooth muscles. A naturally occurring IL-13 polymorphism, R110Q, is strongly associated with increased total serum IgE levels and asthma. In the present study, we aimed to determine whether the IL-13 R110Q variant would display different biochemical properties or altered functions in comparison with wild-type (WT) IL-13 in cultured human bronchial smooth muscle cells (hBSMCs). METHODS: Culture supernatants and cell proteins were collected from cultured hBSMCs that were treated with 50 ng/mL IL-13 or IL-13 R110Q for 24 hours. Eotaxin released into hBSMC culture medium was determined by ELISA. The expression levels of the high-affinity IgE receptor (FcepsilonRI) alpha-chain, smooth muscle-specific actin alpha chain (alpha-SMA), smooth muscle myosin heavy chain (SmMHC), and calreticulin in the cells were measured on Western blots. RESULTS: Compared with WT IL-13, treatment with the IL-13 R110Q variant resulted in a significant increase in eotaxin release as well as significant, although modest, increases in the expression levels of alpha-SMA, SmMHC, calreticulin, and FcepsilonRI alpha-chain. CONCLUSIONS: The results of the present study suggenst that the IL-13 R110Q variant may enhance enhanced functional activities in hBSMCs.

IL-13 R110Q, a Naturally Occurring IL-13 Polymorphism, Confers Enhanced Functional Activity in Cultured Human Bronchial Smooth Muscle Cells

PURPOSE: Interleukin (IL)-13, a Th2-type cytokine, plays a pivotal role in the pathogenesis of asthma through its direct effects on airway smooth muscles. A naturally occurring IL-13 polymorphism, R110Q, is strongly associated with increased total serum IgE levels and asthma. In the present study, we aimed to determine whether the IL-13 R110Q variant would display different biochemical properties or altered functions in comparison with wild-type (WT) IL-13 in cultured human bronchial smooth muscle cells (hBSMCs). METHODS: Culture supernatants and cell proteins were collected from cultured hBSMCs that were treated with 50 ng/mL IL-13 or IL-13 R110Q for 24 hours. Eotaxin released into hBSMC culture medium was determined by ELISA. The expression levels of the high-affinity IgE receptor (FcepsilonRI) alpha-chain, smooth muscle-specific actin alpha chain (alpha-SMA), smooth muscle myosin heavy chain (SmMHC), and calreticulin in the cells were measured on Western blots. RESULTS: Compared with WT IL-13, treatment with the IL-13 R110Q variant resulted in a significant increase in eotaxin release as well as significant, although modest, increases in the expression levels of alpha-SMA, SmMHC, calreticulin, and FcepsilonRI alpha-chain. CONCLUSIONS: The results of the present study suggenst that the IL-13 R110Q variant may enhance enhanced functional activities in hBSMCs.

Dexamethasone Attenuates PDGF- and TGF-beta-enhanced Vascular Endothelial Growth Factor Production in Cultured Human Bronchial Smooth Muscle Cells / 소아알레르기및호흡기학회지

PURPOSE: Human bronchial smooth muscle cell(HBSMC) plays an important role in the remodeling of the airways in asthma. Vascular endothelial growth factor(VEGF) is a multifunctional cytokine, which induces edema, angiogenesis, vascular remodeling, mucus metaplasia, subepithelial fibrosis, and antigen-induced Th2 inflammation. Transforming growth factor-beta(TGF-beta) is a growth modulator of HBSMC and an important cytokine in airway remodeling. We investigated the effect of dexamethasone on the release of VEGF from HBSMC stimulated with platelet-derived growth factor(PDGF) and TGF-beta. METHODS: HBSMC cultured in 10 percent FCS-DMEM media was growth-arrested in serum-deprived medium for 48 hours. Dexamethasone and TGF-beta were added and incubated for 16 hours before stimulation with PDGF. After 24 hours of stimulation, culture medium was harvested and stored at -80 degrees C until ELISA for VEGF was performed. RESULTS: The release of VEGF was significantly increased after stimulation with PDGF (P<0.01). The production of VEGF pretreated with TGF-beta before stimulation with PDGF was higher than those without TGF-beta pretreatment(P<0.01). Dexamethasone suppressed the release of VEGF in HBSMC stimulated with PDGF(P<0.01), TGF-beta and PDGF(P<0.01). CONCLUSION: PDGF and TGF-beta may be one of the key mediators in inducing airway remodeling and glucocorticoid, and can be used as useful therapies to prevent airway vascular remodeling by modulating the VEGF on airway smooth muscle cells.

Effect of Shenmai injection on tone of human isolated bronchial smooth muscle / 中华中医药杂志

0.05),but the contraction of the bronchial rings caused by histamine,potassium chloride,intracellular Ca2+ release and extracellular Ca2+ influx was signifi cantly inhibited by Shenmai injection(compare with those of control,P

Effect of morphine on the Kv channel in bronchial smooth muscle in guinea-pigs / 中国药理学通报

Aim To explore the effect of the morphine(MP) on the Kv channel in bronchial smooth muscle in guinea-pigs (GPBSM) and expressions of the Kv 1.5 mRNA and protein . Methods The effect of the MP on the Kv channel was studied by patch clamp recording in GPBSM and expressions of the Kv 1.5 mRNA and protein were measured by Western-blot and RT-PCR techniques. Results Morphine remarkably restrained the Kv channel current and the expression of the Kv 1.5 mRNA and protein in cultured GPBSM (n=6, each group, P