cyclin-d1 and Airway-Remodeling

Abnormal airway smooth muscle cell (ASMC) proliferation and migration contribute significantly to increased ASM mass associated with asthma. MicroRNA (miR)-638 is a primate-specific miRNA that plays important roles in development, DNA damage repair, hematopoiesis, and tumorigenesis. Although it is highly expressed in ASMCs, its function in ASM remodeling remains unknown. In the current study, we found that in response to various mitogenic stimuli, including platelet-derived growth factor-two B chains (PDGF-BB), transforming growth factor β1, and fetal bovine serum, the expression of miR-638, as determined by quantitative real-time polymerase chain reaction (qRT-PCR), was significantly downregulated in the proliferative human ASMCs. Both gain- and loss-of-function studies were performed to study the role of miR-638 in ASMC proliferation and migration. We found that adenovirus-mediated miR-638 overexpression markedly inhibits ASMC proliferation and migration, while ablation of miR-638 by anti-miR-638 markedly increases cell proliferation and migration, as determined by WST-8 proliferation and scratch wound assays. Dual-luciferase reporter assay, qRT-PCR, and immunoblot analysis were used to investigate the effects of miR-638 on the expression of the downstream target genes in ASMCs. Our results demonstrated that miR-638 overexpression significantly reduced the expression of downstream target cyclin D1 and NOR1, both of which have been shown to be essential for cell proliferation and migration. Together, our study provides the first in vitro evidence highlighting the antiproliferative and antimigratory roles of miR-638 in human ASMC remodeling and suggests that targeted overexpression of miR-638 in ASMCs may provide a novel therapeutic strategy for preventing ASM hyperplasia associated with asthma.

Topics: Airway Remodeling; Asthma; Becaplermin; Cell Movement; Cell Proliferation; Cyclin D1; Gene Expression Regulation; Humans; Membrane Transport Proteins; MicroRNAs; Myocytes, Smooth Muscle

The aims of the present study were to examine the signaling mechanisms for transforming growth factor-β1 (TGF-β1)-induced rat airway smooth muscle cells (ASMCs) proliferation and migration and to determine the effect of lipoxin A4 (LXA4) on TGF-β1-induced rat ASMCs proliferation and migration and its underlying mechanisms. TGF-β1 upregulated transcriptional coactivator Yes-associated protein (YAP) expression by activating Smad2/3 and then upregulated cyclin D1, leading to rat ASMCs proliferation and migration. This effect was reversed after treatment with the TGF-β1 receptor inhibitor SB431542. YAP is a critical mediator of TGF-β1-induced ASMCs proliferation and migration. Knockdown of YAP disrupted the pro-airway remodeling function of TGF-β1. Preincubation of rat ASMCs with LXA4 blocked TGF-β1-induced activation of Smad2/3 and changed its downstream targets, YAP and cyclin D1, resulting in the inhibition of rat ASMCs proliferation and migration. Our study suggests that LXA4 suppresses Smad/YAP signaling to inhibit rat ASMCs proliferation and migration and therefore has potential value in the prevention and treatment of asthma by negatively modulating airway remodeling.

Topics: Airway Remodeling; Animals; Cell Proliferation; Cyclin D1; Myocytes, Smooth Muscle; Rats; Transforming Growth Factor beta1

Airway remodeling in asthma involves the hyperproliferation of airway smooth muscle (ASM) cells. However, the molecular signals that regulate ASM growth are not completely understood. Gq-coupled G protein-coupled receptor and receptor tyrosine kinase signaling regulate ASM cell proliferation via activation of phospholipase C, generation of inositol triphosphate (IP

Topics: Airway Remodeling; Animals; Asthma; Cell Proliferation; Cyclin D1; Diacylglycerol Kinase; Diglycerides; Humans; Inositol; Mice; Mitogens; Myocytes, Smooth Muscle; Phosphatidic Acids; Platelet-Derived Growth Factor; Protein-Tyrosine Kinases; Receptors, G-Protein-Coupled; TOR Serine-Threonine Kinases; Type C Phospholipases

The purpose of this study was to establish a rat asthma model and extract MUC5AC to explore the mechanism of mucin 5AC (MUC5AC) signaling pathway regulating the function of asthmatic airway smooth muscle cells (ASMC) and participating in asthmatic airway remodeling. Western blot was used to detect β-catenin (β-catenin), glycogen synthase kinase-3β (GSK-3β), proto-oncogene MUC5AC and cyclin D1 (cyclin D1) in MUC5AC of asthmatic and normal groups. After inhibiting the interaction between β-catenin and transcription cofactor p300 / CBP in ASMC of the asthma group and control group, the cell viability and cycle changes of ASMC were detected by the CCK-8 method and flow cytometry. After inhibiting the activity of P38 mitogen-activated protein kinase (MAPK), the protein expression changes of c-Myc and cyclin D1 were detected by Western blot. Results showed that comprehensive HE staining results of lung tissue sections indicate that the experimental rat model of asthma airway remodeling was successfully established. Compared with the control group, 100 fxmol and L1 Efaroxan promoted insulin secretion (P <0.01), and administration of the MUC5AC antagonist KU14R significantly inhibited the effect of MUC5AC.Western blot showed that the protein expression levels of β-catenin, c-Myc and cyclin D1 in ASMC of the obese asthma group were significantly higher than those of the control group (P <0.05), while the protein expression level of GSK-3β was lower than Control group (P <0.05). After inhibiting the interaction between β-catenin and p300 / CBP, the decrease in cell viability and the degree of cell cycle change of ASMC in the asthma group were more obvious than those in the control group (P <0.05). After inhibiting the activity of P38 MAPK, the expressions of the target proteins c-Myc and cyclin D1 in the MUC5AC signaling pathway in ASMC model rats and control rats were down-regulated, and the difference was statistically significant (P <0.05). The conclusion was that the Wnt/β-catenin signaling pathway can regulate the proliferation and differentiation of ASMC by up-regulating the expression level of cMyc. Cyclin D1 interacts with the MAPK signaling pathway, thereby affecting the function of ASMC and participating in asthma airway remodeling.

Topics: Airway Remodeling; Animals; Asthma; beta Catenin; Cell Proliferation; Cyclin D1; Glycogen Synthase Kinase 3 beta; Mucin 5AC; Obesity; p38 Mitogen-Activated Protein Kinases; Rats; Wnt Signaling Pathway

Abnormal proliferation of airway smooth muscle cells (ASMCs) is a hallmark of airway remodeling. Platelet-derived growth factor (PDGF) is known to be a major stimulus inducing the proliferation of ASMCs. It has been reported that triptolide demonstrates protective effects against airway remodeling. In this study, we investigated the antiproliferative effects of triptolide on PDGF-induced ASMCs and its underlying mechanisms. Cell proliferation was determined using the Cell Counting Kit-8 (CCK-8) assay. Flow cytometry was used to study the influence of triptolide on cell cycle and apoptosis. Quantitative real-time PCR and Western blot analysis were employed to detect the expression of proliferating cell nuclear antigen (PCNA), cyclinD1 and cyclin dependent kinase 4 (CDK4). Proteins involved in the protein kinase B (AKT) and nuclear factor kappa B (NF-κB) signaling pathways were evaluated using Western blot analysis. Triptolide could significantly inhibit cell proliferation, induce cell cycle arrest in the G0/G1 phase, and reduce the expression of PCNA, cyclinD1, and CDK4 in PDGF-treated ASMCs. Levels of phosphorylated AKT, p65 and NF-κB inhibitor α (IκBα) stimulated by the presence of PDGF were markedly suppressed after triptolide treatment. Moreover, triptolide cotreatment with the phosphatidylinositol 3 kinase (PI3k) inhibitor, 2-(4-morpholinyl)-8-phenylchromone (LY294002), could further suppress the proliferation, NF-κB activation and cyclinD1 expression. Similar results were observed after triptolide cotreatment with the NF-κB inhibitor, ammonium pyrrolidinedithiocarbamate (PDTC). Our results suggest that triptolide could inhibit the PDGF-induced proliferation of ASMCs through G0/G1 cell cycle arrest and suppression of the AKT/NF-κB/cyclinD1 signaling pathway.

Topics: Airway Remodeling; Animals; Asthma; Bronchi; Cell Proliferation; Cells, Cultured; Chromones; Cyclin D1; Diterpenes; Epoxy Compounds; G1 Phase Cell Cycle Checkpoints; Humans; Morpholines; Myocytes, Smooth Muscle; NF-kappa B; Phenanthrenes; Phosphatidylinositol 3-Kinases; Phosphorylation; Platelet-Derived Growth Factor; Primary Cell Culture; Proto-Oncogene Proteins c-akt; Pyrrolidines; Rats; Signal Transduction; Thiocarbamates

YKL-40 is a chitinase-like protein which was significantly elevated in asthma patients and related closely to asthma severity and airway remodeling. Airway remodeling in asthma involves complicated physical and pathological processes, including increased airway smooth muscle mass due to proliferation, migration of airway smooth muscle cells, epithelial-mesenchymal transition (EMT) and sub-epithelial fibrosis. However, the precise effect and underlying mechanism of YKL-40 in this pathological alteration remained unelucidated. In this study, we demonstrated that YKL-40 could promote asthma airway remodeling by increasing airway smooth muscle mass, inducing EMT and sub-epithelial fibrosis. Furthermore, we identified that FAK and MAPK signaling pathways are activated in the process. Inhibiting FAK or MAPK pathway could significantly ameliorate airway remodeling induced by excessive secretion of YKL-40

Topics: Airway Remodeling; Animals; Asthma; Cell Movement; Cell Proliferation; Chitinase-3-Like Protein 1; Cyclin D1; Cyclin-Dependent Kinases; Disease Models, Animal; Epithelial Cells; Epithelial-Mesenchymal Transition; Fibrosis; Focal Adhesion Protein-Tyrosine Kinases; Humans; MAP Kinase Signaling System; Mice, Inbred C57BL; Myocytes, Smooth Muscle; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-akt; src-Family Kinases

Airway smooth muscle (ASM) cell hyperplasia contributes to airway wall remodeling (AWR) in asthma. Glucocorticoids, which are used as first-line therapy for the treatment of inflammation in asthma, have limited impact on AWR, and protracted usage of high doses of glucocorticoids is associated with an increased risk of side effects. Moreover, patients with severe asthma often show reduced sensitivity to glucocorticoids. Artesunate, a semisynthetic artemisinin derivative used to treat malaria with minimal toxicity, attenuates allergic airway inflammation in mice, but its impact on AWR is not known. We examined the effects of artesunate on ASM proliferation in vitro and in vivo. Primary human ASM cells derived from nonasthmatic donors were treated with artesunate before mitogen stimulation. Artesunate reduced mitogen-stimulated increases in cell number and cyclin D1 protein abundance but had no significant effect on ERK1/2 phosphorylation. Artesunate, but not dexamethasone, inhibited phospho-Akt and phospho-p70(S6K) protein abundance. Artesunate, but not dexamethasone, inhibited mitogen-stimulated increases in cell number, cyclin D1, and phospho-Akt protein abundance on ASM cells derived from asthmatic donors. In a murine model of allergic asthma, artesunate reduced the area of α-smooth muscle actin-positive cells and decreased cyclin D1 protein abundance. Our study provides a basis for the future development of artesunate as a novel anti-AWR agent that targets ASM hyperplasia via the PI3K/Akt/p70(S6K) pathway and suggests that artesunate may be used as combination therapy with glucocorticoids.

Topics: Airway Remodeling; Animals; Antimalarials; Artemisinins; Artesunate; Asthma; Cell Proliferation; Cells, Cultured; Cyclin D1; Dexamethasone; Female; Glucocorticoids; Humans; Mice; Muscle, Smooth; Myocytes, Smooth Muscle; Phosphatidylinositol 3-Kinases; Respiratory System

To investigate the effects of peroxisome proliferator-activated receptor-gamma (PPARγ) agonist rosiglitazone on the expression of cyclin D1 in lung tissue, and the proliferation of airway smooth muscle cells (ASMCs) in mice with bronchial asthma.. Thirty clean BALB/c mice were randomly divided into control group (n = 10), asthma group (n = 10), and rosiglitazone treatment group (n = 10). A mouse model of asthma was established by ovalbumin (OVA) sensitization and challenge. The treatment group received rosiglitazone (5 mg/kg) by gavage 1 hour before each challenge and the control group received saline instead of OVA sensitization and challenge. Leukocytes and eosinophils in bronchoalveolar lavage fluid (BALF) were counted under a microscope. Airway structural changes were observed by hematoxylin-eosin staining. Protein and mRNA expression levels of cyclin D1 were measured by immunohistochemical staining and RT-PCR. Perimeter of the basement membrane (Pbm), total bronchial wall area (WAt), airway smooth muscle area (WAm), and number of nuclei in ASMCs (N) were determined using image analysis software, and WAt/Pbm, WAm/Pbm, and N/Pbm were calculated.. Compared with the control group, the asthma group showed significant increases in the total number of leukocytes and percentage of eosinophils in BALF, as well as in the mRNA and protein expression of cyclin D1, but changes in these indices were significantly reduced in the rosiglitazone treatment group (P < 0.05). In addition, compared with the control group, the asthma group had significantly increased WAt/Pbm, WAm/Pbm, and N/Pbm, but rosiglitazone significantly decreased these ratios (P < 0.05).. Rosiglitazone may delay the process of airway remodeling by inhibiting the proliferation of ASMCs, so it can be used for preventing and treating chronic asthma.

Topics: Airway Remodeling; Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Cell Proliferation; Cyclin D1; Female; Lung; Mice; Mice, Inbred BALB C; Myocytes, Smooth Muscle; PPAR gamma; RNA, Messenger; Rosiglitazone; Thiazolidinediones

It is well established that hyperplasia and decreased apoptosis of airway smooth muscle cells (ASMCs) play an important role in the asthmatic airway remodeling. Tumor suppressor PTEN gene with phosphatase activity plays an important regulatory role in embryonic development, cell proliferation, and apoptosis, cell cycle regulation, migration (invasion) of the cytoskeleton. We hypotheses that PTEN gene could affect the growth and viability of ASMCs through the regulation of PI3K/Akt, MAPK, and cell cycle-related gene expression. We constructed a recombinant adenovirus to transfect ASMCs. Cells were divided into the overexpression of PTEN gene group (Ad-PTEN-GFP), negative control group (Ad-GFP), and blank control group (DMEM). The cell apoptosis of ASMCs were evaluated by Hoechst-33342 staining and PE-7AAD double-labeled flow cytometry. The cell cycle distribution was observed by flow cytometry with PI staining. The expression of PTEN, p-Akt, total-Akt, p-ERK1/2, total-ERK1/2, cleaved-Caspases-3, Caspases-9, p21, and Cyclin D1 were tested by the Western blotting. Our study revealed that overexpression of PTEN gene did not induce apoptosis of human ASMCs cultured in vitro. However, overexpression of PTEN inhibited proliferation of human ASMCs cultured in vitro and was associated with downregulation of Akt phosphorylation levels, while did not affect ERK1/2 phosphorylation levels. Moreover, overexpression of PTEN could induce ASMCs arrested in the G0/G1 phase through the downregulation of Cyclin D1 and upregulation of p21 expressions.

Topics: Adenoviridae; Airway Remodeling; Apoptosis; Asthma; Caspase 3; Caspase 9; Cell Proliferation; Cell Shape; Cell Survival; Cells, Cultured; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; G1 Phase Cell Cycle Checkpoints; Gene Expression; Humans; Lung; Myocytes, Smooth Muscle; Phosphorylation; Protein Processing, Post-Translational; Proto-Oncogene Proteins c-akt; PTEN Phosphohydrolase; Transfection

Chronic hypoxia causes pulmonary vascular remodeling leading to pulmonary hypertension (PH) and right ventricle (RV) hypertrophy. Aberrant expression of microRNA (miRNA) is closely associated with a number of pathophysiologic processes. However, the role of miRNAs in chronic hypoxia-induced pulmonary vascular remodeling and PH has not been well characterized. In this study, we found increased expression of miR-21 in distal small arteries in the lungs of hypoxia-exposed mice. Putative miR-21 targets, including bone morphogenetic protein receptor (BMPR2), WWP1, SATB1, and YOD1, were downregulated in the lungs of hypoxia-exposed mice and in human pulmonary artery smooth muscle cells (PASMCs) overexpressing miR-21. We found that sequestration of miR-21, either before or after hypoxia exposure, diminished chronic hypoxia-induced PH and attenuated hypoxia-induced pulmonary vascular remodeling, likely through relieving the suppressed expression of miR-21 targets in the lungs of hypoxia-exposed mice. Overexpression of miR-21 enhanced, whereas downregulation of miR-21 diminished, the proliferation of human PASMCs in vitro and the expression of cell proliferation associated proteins, such as proliferating cell nuclear antigen, cyclin D1, and Bcl-xL. Our data suggest that miR-21 plays an important role in the pathogenesis of chronic hypoxia-induced pulmonary vascular remodeling and also suggest that miR-21 is a potential target for novel therapeutics to treat chronic hypoxia associated pulmonary diseases.

Topics: Airway Remodeling; Animals; Apoptosis; bcl-X Protein; Bone Morphogenetic Protein Receptors, Type II; Cell Cycle Proteins; Cell Line; Cell Proliferation; Cyclin D1; Down-Regulation; Humans; Hypertension, Pulmonary; Hypertrophy, Right Ventricular; Hypoxia; Lung; Matrix Attachment Region Binding Proteins; Mice; MicroRNAs; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Pulmonary Artery; Ubiquitin-Protein Ligases; Up-Regulation

To investigate the effects of H(2) Relaxin (Relaxin) on airway remodeling and the expression of cyclin D(1) in a murine model of chronic asthma.. Forty BALB/c mice were randomly divided into 4 groups:a normal control group, an asthma group, a vehicle control group and a relaxin treatment group, with 10 mice in each. The mice were sensitized and challenged with ovalbumin (OVA) to establish the chronic asthmatic model. The vehicle control group and the relaxin treatment group were subcutaneously injected with saline and relaxin (0.25 mg × kg(-1)× d(-1))respectively. Alteration of the airway inflammation and collagen deposition were observed by haematoxylin-eosin (HE) and Masson staining. Hydroxyproline in the lung was measured by enzyme linked immunosorbent assay (ELISA). The expression of α-smooth muscle actin (α-SMA) in lungs was evaluated by immunohistochemistry. The protein expression and the mRNA of cyclin D(1) were detected by Western blot and RT-PCR respectively.. There were inflammatory cell infiltration, airway stenosis, bronchial smooth muscle hypertrophy and increased collagen deposition in the asthmatic group and the vehicle control group; but these changes were significantly ameliorated in the relaxin treatment group. The area of the α-SMA-stained smooth muscle layer in the asthmatic group and the vehicle control group was significantly greater than that in the control group (all P < 0.05), while administration of relaxin decreased the α-SMA immunostained area (all P < 0.05). The lung hydroxyproline content in the asthmatic and the vehicle groups [(0.68 ± 0.10) mg/g lung tissue, (0.67 ± 0.10) mg/g lung tissue] was significantly greater than that in the control group [(0.26 ± 0.05) mg/g lung tissue] (q = 16.61, 16.01 respectively, all P < 0.01). In contrast, treatment with relaxin significantly reduced the lung hydroxyproline content [(0.40 ± 0.06) mg/g lung tissue] compared with aforementioned 2 groups (q = 10.88, 10.26 respectively, all P < 0.05). The results of the Western blot analysis showed that the expression level of cyclin D(1) in the asthmatic and the vehicle groups [(1.38 ± 0.18), (1.50 ± 0.10)] was higher than that in the control group (0.38 ± 0.10) (q = 13.00, 14.65 respectively, all P < 0.05), while it was significantly decreased in the relaxin group (0.72 ± 0.13) (q = 8.51, 10.16 respectively, all P < 0.05). There were no differences in all of the parameters between the asthmatic group and the vehicle group (P > 0.05).. Relaxin alleviated airway inflammation, airway smooth muscle thickening and airway remodelling in a murine model of chronic asthma, partially by down-regulating the expression level of cyclin D(1).

Topics: Actins; Airway Remodeling; Animals; Asthma; Cyclin D1; Female; Lung; Mice; Mice, Inbred BALB C; Relaxin

A major feature of chronic obstructive pulmonary disease (COPD) is airway remodelling, which includes an increased airway smooth muscle (ASM) mass. The mechanisms underlying ASM remodelling in COPD are currently unknown. We hypothesized that cigarette smoke (CS) and/or lipopolysaccharide (LPS), a major constituent of CS, organic dust and gram-negative bacteria, that may be involved in recurrent airway infections and exacerbations in COPD patients, would induce phenotype changes of ASM.. To this aim, using cultured bovine tracheal smooth muscle (BTSM) cells and tissue, we investigated the direct effects of CS extract (CSE) and LPS on ASM proliferation and contractility.. Both CSE and LPS induced a profound and concentration-dependent increase in DNA synthesis in BTSM cells. CSE and LPS also induced a significant increase in BTSM cell number, which was associated with increased cyclin D1 expression and dependent on activation of ERK 1/2 and p38 MAP kinase. Consistent with a shift to a more proliferative phenotype, prolonged treatment of BTSM strips with CSE or LPS significantly decreased maximal methacholine- and KCl-induced contraction.. Direct exposure of ASM to CSE or LPS causes the induction of a proliferative, hypocontractile ASM phenotype, which may be involved in airway remodelling in COPD.

Topics: Airway Remodeling; Animals; Cattle; Cell Proliferation; Cells, Cultured; Cyclin D1; DNA Replication; Dose-Response Relationship, Drug; Flavonoids; Imidazoles; Isometric Contraction; Lipopolysaccharides; Methacholine Chloride; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Muscle, Smooth; Myocytes, Smooth Muscle; p38 Mitogen-Activated Protein Kinases; Phenotype; Phosphorylation; Potassium Chloride; Protein Kinase Inhibitors; Pulmonary Disease, Chronic Obstructive; Pyrimidines; Smoke; Smoking; Time Factors; Tissue Culture Techniques; Trachea

To investigate the expression variations of connective tissue growth factor (CTGF) and cyclin D1 in pulmonary vasculature in rats exposed to cigarette smoke and their roles in pulmonary vascular remodeling.. Twenty-four male Wistar rats were randomly divided into 4 groups: 1 control group (C group) and 3 smoke exposure groups (S2w, S4w, S8w group). Arterial partial pressure of oxygen was measured. Pulmonary artery remodeling was observed by Hematoxylin-Eosin staining and the percentage of muscularised small pulmonary arteries. Immunohistochemistry methods were performed to observe CTGF and cyclin D1 expressions in pulmonary artery smooth muscle. Real time quantitative RT-PCR and Western blot analysis were used for detection of mRNA and protein expressions in pulmonary artery smooth muscle.. There was no significant difference in arterial partial pressure of oxygen among all groups. The percentage of muscularised small vessels and W/T were significantly increased in S2w, S4w and S8w group compared to control group (P < 0.05). Compared to control group, significant increases of CTGF and cyclinD1 expressions in smoke exposure groups were observed (P < 0.05). The expressions of CTGF and cyclinD1 were significantly positively correlated with the severity of pulmonary vascular muscularization, and there was statistically positive correlation between the expression of CTGF and cyclinD1.. CTGF and cyclinD1 expressions significantly were upregulated in pulmonary arteries from rats exposed to cigarette smoke (2-8w) and there was a significant positive correlation between their expressions. Their expression variations may be associated with abnormal proliferation of pulmonary artery smooth muscle cells induced by cigarette smoke.

Topics: Airway Remodeling; Animals; Connective Tissue Growth Factor; Cyclin D1; Inhalation Exposure; Lung; Male; Muscle, Smooth, Vascular; Rats; Rats, Wistar; Smoke Inhalation Injury; Smoking

To evaluate the expression of cyclin D(1) in asthmatic mouse lungs, and to explore the role of cyclin D(1) in bronchial asthma and airway remodeling.. Forty BALB/c mice were randomized to group A (normal), group B (sensitized for 2 weeks), group C (sensitized for 4 weeks) and group D (sensitized for 8 weeks), 10 mice each group. The mice were sensitized with 10% ovalbumin and challenged with 1% ovalbumin to establish the asthmatic model. The number of eosinophils and the cell percentages in bronchoalveolar lavage fluid (BALF) were counted by cytology method. Pulmonary functions were measured to evaluate the resistance of expiration. Airway inflammation and eosinophil infiltration were evaluated by HE staining, and the airway wall thickness (WAt/Pi), smooth muscle thickness (WAm/Pi) and smooth nucleus counts (N/Pi) were quantified by computer-assisted image analysis system. The mRNA expression of cyclin D(1) was measured by RT-PCR and Real-time PCR. The protein expression of cyclin D(1) was assayed by Western blot. The correlation between airway resistance of expiration and the expression of cyclin D(1) was studied.. The eosinophil count and differential in BALF of group B, C, and D [(42.6 +/- 0.9) x 10(4)/L, (54.7 +/- 1.4) x 10(4)/L, (44.8 +/- 2.4) x 10(4)/L] were higher than those of group A (3.4 +/- 0.5) x 10(4)/L (q = 79.75, 91.42, 84.82, all P < 0.01). The airway resistance of expiration after challenge with 45 microg/kg acetylcholine in group B, C, and D [(5.27 +/- 0.16) cmxL(-1)xmin(-1), (6.68 +/- 0.20) cmxL(-1)xmin(-1), (7.14 +/- 0.41) cmxL(-1)xmin(-1)] was higher than that in group A [(4.11 +/- 0.15) cmxL(-1)xmin(-1), q = 5.58, 6.39, 7.11, all P < 0.05]. Eosinophil infiltration, cilium loss, formation of mucus plug and smooth muscle cell layer thickening were observed in group B, C, and D. The morphological changes of the airways became evident following airway remodeling. WAm/Pi in group B, C, and D (2.8 +/- 0.6, 4.8 +/- 0.6, 6.4 +/- 0.7) were higher than in group A (2.4 +/- 0.4, q = 6.40, 8.28, 9.27, all P < 0.05), and WAt/Pi in group B, C, and D (6.4 +/- 0.8, 8.3 +/- 1.2, 9.3 +/- 1.0) were higher than in group A (5.6 +/- 1.0, q = 2.80, 4.83, 6.37, all P < 0.05). The protein expression of Cyclin D(1) in group B, C, and D (0.587 +/- 0.015, 0.808 +/- 0.029, 0.826 +/- 0.022) were higher than in group A (0.404 +/- 0.016, q = 5.87, 8.08, 8.26, all P < 0.01). There was a positive correlation between the expression of cyclin D(1) and airway resistance (r = 0.83, P < 0.05).. The expression of cyclin D(1) in the asthmatic mouse lung was increased, and positively correlated to airway reactivity. Cyclin D(1) might be essential in the airway remodeling of asthma through ERK signaling pathway.

Topics: Airway Remodeling; Animals; Asthma; Cyclin D1; Eosinophils; Female; Lung; Mice; Mice, Inbred BALB C