Interleukin-13 In Bronchoalveolar Lavage Fluid Research Articles

Suppression of autophagy through JAK2/STAT3 contributes to the therapeutic action of rhynchophylline on asthma

BackgroundAsthma is a chronic inflammatory disease characterized by airway remodeling and inflammation. Rhynchophylline is a kind of indole alkaloid isolated from Uncaria rhynchophylla. Here we investigated the effect of rhynchophylline on autophagy in asthma.MethodsA mice model of asthma was established by ovalbumin challenge. Histopathological changes were assessed by hematoxylin-eosin staining, periodic acid-schiff staining and Masson staining. The levels of IgE in serum, interleukin-6 and interleukin-13 in bronchoalveolar lavage fluid, as well as the activities of superoxide dismutase and catalase in lung tissues were detected. The expression of autophagy-related genes and Janus kinase (JAK) 2/ signal transducer and activator of transcription (STAT) 3 signal was detected by western blot and immunofluorescence. Airway smooth muscle cells (ASMCs) were isolated, and the effect rhynchophylline on autophagy in ASMCs was explored.ResultsOur data showed that rhynchophylline treatment alleviated inflammation, airway remodeling, and oxidative stress in asthma. In addition, autophagy, which was implicated in asthma, was suppressed by rhynchophylline with decreased level of autophagy-related proteins. Furthermore, rhynchophylline suppressed the JAK2/STAT3 signaling pathway, which was activated in asthma. In vitro study showed that rhynchophylline suppressed ASMC autophagy through suppressing the activation of JAK2/STAT3 signal.ConclusionsOur study demonstrated that rhynchophylline can alleviate asthma through suppressing autophagy in asthma, and that JAK2/STAT3 signal was involved in this effect of rhynchophylline. This study indicates that rhynchophylline may become a promising drug for the treatment of asthma.

Bone marrow mesenchymal stem cells protect lungs from smoke inhalation injury by differentiating into alveolar epithelial cells via Notch signaling

To examine the protective effect of transplanting bone marrow mesenchymal stem cells (BMSCs) in treating lung injuryinduced by smoke exposure and to investigate the underlying mechanisms of this protection. SD rats were randomlydivided into four groups: normal group, normal +BMSCGFP group, smoke group, and smoke +BMSCGFP group. Todetect lung injury, we measured arterial blood gas, the wet-to-dry weight ratio, and levels of interleukin-1b, tumor necrosisfactor-a, interleukin-10, and interleukin-13 in bronchoalveolar lavage fluid and lung tissues. We also conductedhistopathology examinations. The protein markers of alveolar epithelial cells were measured to determine the BMSCdifferentiation. The protein levels of Notch1, Jagged-1, and Hes-1 also were detected. In the present study, BMSCtransplantation significantly decreased the wet-dry weight ratio of the lung, reduced the production of inflammatorymediators, and alleviated lung injury simply through differentiating into alveolar type II cells and alveolar type I cells. Western blot analysis confirmed that the protein expression of Notch-1, Jagged-1, and Hes-1 increased significantly aftersystemic BMSC transplantation. No significant difference was observed between the normal group and the nor-mal +BMSCGFP group. Our findings indicate that systemic transplantation of BMSCs alleviated lung injury induced bysmoke exposure, which may be associated with BMSCs' ability to differentiate into alveolar-type cells via the Notchsignaling pathway.

Ghrelin Ameliorates Asthma by Inhibiting Endoplasmic Reticulum Stress

BackgroundThis study aimed to confirm the ameliorative effect of ghrelin on asthma and investigate its mechanism. Materials and MethodsThe murine model of asthma was induced by ovalbumin (OVA) treatment and assessed by histological pathology and airway responsiveness to methacholine. The total and differential leukocytes were counted. Tumor necrosis factor α, interferon γ, interleukin-5 and interleukin-13 levels in bronchoalveolar lavage fluid were quantified by commercial kits. The protein levels in pulmonary tissues were measured by Western blot analysis. ResultsGhrelin ameliorated the histological pathology and airway hyperresponsiveness in the OVA-induced asthmatic mouse model. Consistently, OVA-increased total and differential leukocytes and levels of tumor necrosis factor α, interferon γ, interleukin-5 and interleukin-13 in bronchoalveolar lavage fluid were significantly attenuated by ghrelin. Ghrelin prevented the increased protein levels of the endoplasmic reticulum stress markers glucose regulated protein 78 and CCAAT/enhancer binding protein homologous protein and reversed the reduced levels of p-Akt in asthmatic mice. ConclusionsGhrelin might prevent endoplasmic reticulum stress activation by stimulating the Akt signaling pathway, which attenuated inflammation and ameliorated asthma in mice. Ghrelin might be a new target for asthma therapy.

Effect of sevoflurane on autophagy in lung tissues of asthmatic mice

Objective To investigate the effect of sevoflurane on autophagy in lung tissues of asthmatic mice. Methods Twenty-four SPF female C57BL/6 mice, aged 6-8 weeks, weighing 23-27 g, were randomly divided into 3 groups (n=8 each) using a random number table: control group, asthma group and sevoflurane group.The mice were sensitized with intraperitoneal ovalbumin 10 μg and 1 mg potassium aluminium sulfate in 0.5 ml of normal saline (NS) on 1st day of establishment of the model, and asthma was challenged with aerosolized 1% ovalbumin in 50 ml of NS for 30 min once a day for 7 consecutive days starting from 15th day in asthma group.Asthma was induced using the method mentioned above, and the sensitized animals inhaled 3% sevoflurane for 1 h after each challenge in sevoflurane group.In control group, potassium aluminium sulfate 1 mg in 0.5 ml of NS was intraperitoneally injected on 1st day of establishment of the model, and the mice were exposed to aerosolized 0.9% NS 50 ml for 30 min once a day for 7 consecutive days starting from 15th day.At 24 h after the last challenge, the lung was lavaged, and broncho-alveolar lavage fluid (BALF) was collected for determination of the inflammatory cell count and concentrations of interleukin-13 (IL-13) and IL-10 in BALF (by enzyme-linked immunosorbent assay). The lung tissues were taken for examination of the inflammatory cell infiltration and damage to the bronchial epithelia and to the structure of alveolus and for determination of Beclin-1 and microtubule-associated protein 1 light chain 3 (LC3) expression (by Western blot). Results Compared with control group, the inflammatory cell count and concentrations of IL-13 and IL-10 in BALF were significantly increased, and the expression of Beclin-1 and LC3 in lung tissues was significantly up-regulated in asthma group (P<0.05). Compared with asthma group, the inflammatory cell count and concentrations of IL-13 in BALF were significantly decreased, the IL-10 concentration in BALF was significantly increased, and the expression of Beclin-1 and LC3 in lung tissues was significantly down-regulated in sevoflurane group (P<0.05). There was significantly less inflammatory cell infiltration in lung tissues, necrosis and shedding of bronchial epithelial cells and alveolar wall disruption in sevoflurane group. Conclusion The mechanism by which sevoflurane inhibits airway inflammation is related to inhibition of autophagy in asthmatic mice. Key words: Asthma; Anesthetics, inhalation; Autophagy; Lung

Effects of chrysin on steroid-resistant asthma in a murine model

To explore the therapeutic effects of chrysin for steroid-resistant asthma in a murine model. Lipopolysaccharide (LPS) was used to induce steroid-resistant asthma in a murine model sensitized and challenged with ovalbumin (OVA). Forty-eight female BALB/c mice were randomly divided into 6 groups of control (A), OVA (B), LPS + OVA (C), LPS + OVA + dexamethasone (D), LPS + OVA + chrysin (E) and LPS + OVA + dexamethasone + chrysin (F) (n = 8 each). At Day 1 and 14, group A received an intraperitoneal injection of phosphate buffered saline (PBS); groups B, C, D, E and F had an intraperitoneal injection of a mixture of OVA (20 µg) and aluminum hydroxide for sensitization. At Day 27, groups A and B were intranasally instilled with PBS while groups C, D, E and F had an intranasal instillation of LPS (10 µg). At Days 28, 29 and 30, groups B, C, D, E and F were challenged via airway with 1% OVA in PBS for 30 min and group A with PBS. Group D was intraperitoneally injected with dexamethasone (3 mg/kg) 30 min pre-challenge and PBS one day pre-challenge; group E received an intraperitoneal injection of chrysin (50 mg/kg) at one day and 1h pre-challenge; group F received dexamethasone (3 mg/kg) 30 min pre-challenge and chrysin (50 mg/kg) at one day and 1 h pre-challenge; groups A, B and C had PBS as above. Hematoxylin-eosin (HE) staining and periodic acid-Schiff (PAS) staining of lung tissues were performed to observe the pathologic changes. The total cells in bronchoalveolar lavage fluid (BALF) were counted under microscope. And enzyme-linked immunosorbent assay (ELISA) was applied for detecting interleukin-4 (IL-4) and interleukin-13 (IL-13) in BALF and IgE in sera. The inflammatory infiltration in lung tissue of group F was obviously suppressed compared with that of group C. The numbers of PAS-positive cells per bronchus divided by the total number of epithelial cells in group D, E, F were markedly lower than that in group C ((54.5 ± 6.9)%, (53.3 ± 8.2)%, (23.8 ± 7.0)% vs (71.3 ± 12.2)%, all P < 0.01). The total cells in BALF of group E and F significantly decreased versus that of group C ((1.22 ± 0.23) × 10⁴/ml, (0.98 ± 0.25) × 10⁴/ml vs (2.56 ± 0.18) × 10⁴/ml, both P < 0.01). The levels of IL-4 in group D and F were significantly less than that of group C ((118 ± 7), (124 ± 5) vs (138 ± 6) pg/ml, both P < 0.01). The levels of IL-13 in BALF of group E and F significantly decreased compared with that of group C ((787 ± 57), (484 ± 32) vs (1 121 ± 132) pg/ml, both P < 0.01). The levels of IgE in sera of group D, E, F were significantly lower those of group C ((10 310 ± 494), (10 771 ± 650), (7 529 ± 485) vs (12 618 ± 595) ng/ml, all P < 0.01). Chrysin could improve the therapeutic efficacies of glucocorticoid for steroid-resistant asthma.

Platycodin D Attenuates Airway Inflammation in a Mouse Model of Allergic Asthma by Regulation NF-κB Pathway.

In this work, the anti-asthma potential of platycodin D (PLD) was studied by investigation of its effect to suppress airway inflammation, a murine model of asthma and the possible mechanisms. A total of 50 mice were randomly assigned to five experimental groups: control, ovalbumin (OVA), OVA+dexamethasone (2 mg/kg) and OVA+PLD (40, 80 mg/kg). Airway resistance (Raw) were measured; airway histological studies were evaluated by the hematoxylin and eosin (HE) staining; interleukin-4 (IL-4), interleukin-5(IL-5), and interleukin-13 in bronchoalveolar lavage fluid (BALF) were evaluated by enzyme-linked immunosorbent assay (ELISA); NF-κBp65, p-NF-κBp65, p-IKKα, IKKα, p-IKKβ, p-IкBα, and IкBα of airway were measured by Western blotting. Our study demonstrated that PLD inhibited OVA-induced increases in Raw and eosinophil count in airway; IL-4, IL-5, and IL-13 were recovered in BALF. Histological studies demonstrated that PLD substantially inhibited OVA-induced eosinophilia in airway tissue. Western blotting studies demonstrated that PLD substantially inhibited NF-κB pathway. These findings suggest that PLD may effectively ameliorate the progression of asthma and could be used as a therapy for patients with allergic asthma.

Effects of taraxasterol on ovalbumin-induced allergic asthma in mice

Ethnopharmacological relevanceTaraxasterol was isolated from the Chinese medicinal herb Taraxacum officinale which has been frequently used as a remedy for inflammatory diseases. In the present study, we determined the in vivo protective effect of taraxasterol on allergic asthma induced by ovalbumin (OVA) in mice. Materials and methodsMice were sensitized and challenged with OVA, and were orally treated daily with taraxasterol at 2.5, 5 and 10mg/kg from day 23 to 27 after sensitization. The number of inflammatory cells in bronchoalveolar lavage fluid (BALF) was determined. Th2 cytokine interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-13 (IL-13) production in BALF and OVA-specific immunoglobulin E (IgE) production in sera were measured using ELISA. Histological changes in lung tissues were examined using hematoxylin and eosin (H&E) and periodic acid-Schiff staining (PAS). Airway hyperresponsiveness (AHR) to inhaled methacholine was assessed. ResultsTaraxasterol dramatically decreased the total inflammatory cell and main inflammatory cell counts, reduced the production of Th2 cytokine IL-4, IL-5, IL-13 in BALF and OVA-specific IgE in sera, and suppressed AHR in a dose-dependent manner. Histological studies demonstrated that taraxasterol substantially suppressed OVA-induced inflammatory cells infiltration into lung tissues and goblet cell hyperplasia in airways. ConclusionsThis finding suggests that taraxasterol protects against OVA-induced allergic asthma in mice.

Murine Cytomegalovirus Influences Foxj1 Expression, Ciliogenesis, and Mucus Plugging in Mice with Allergic Airway Disease

We have followed throughout time the development of allergic airway disease (AAD) in both uninfected mice and mice infected intranasally with murine cytomegalovirus (MCMV). Histological evaluation of lung tissue from uninfected mice with AAD demonstrated mucus plugging after 14 and 21 days of ovalbumin-aerosol challenge, with resolution of mucus plugging occurring by 42 days. In MCMV/AAD mice, mucus plugging was observed after 7 days of ovalbumin-aerosol challenge and remained present at 42 days. The level of interleukin-13 in bronchoalveolar lavage fluid from MCMV/AAD mice was decreased compared with AAD mice and was accompanied by increased levels of interferon-gamma. Levels of Muc5A/C, Muc5B, or Muc2 mucin mRNA in the lungs of MCMV/AAD mice were not elevated compared with AAD mice. MCMV was able to infect the airway epithelium, resulting in decreased expression of Foxj1, a transcription factor critical for ciliogenesis, and a loss of ciliated epithelial cells. In addition, an increase in the number of epithelial cells staining positive for periodic acid-Schiff was observed in MCMV/AAD airways. Together, these findings suggest that MCMV infection of the airway epithelium enhances goblet cell metaplasia and diminishes efficient mucociliary clearance in mice with AAD, resulting in increased mucus plugging.