Triamcinolone acetonide induces the autophagy of Ag85B-treated WI-38 cells via SIRT1/FOXO3 pathway.

BACKGROUND: Tracheobronchial stenosis due to tuberculosis (TSTB) seriously threatens the health of tuberculosis patients. The inflammation and autophagy of fibroblasts affect the development of TSTB. Triamcinolone acetonide (TA) can regulate the autophagy of fibroblasts. Nevertheless, the impact of TA on TSTB and underlying mechanism has remained unclear. OBJECTIVE: To study the impact of TA on TSTB and underlying mechanism. MATERIAL AND METHODS: In order to simulate the TSTB-like model in vitro, WI-38 cells were exposed to Ag85B protein. In addition, the cell counting kit (CCK)-8 assay was applied to assess the function of TA in Ag85B-treated WI-38 cells. Quantitative real-time polymerase chain reaction was applied to detect the mRNA level of sirtuin 1 (SIRT1) and forkhead box O3 (FOXO3a), and autophagy-related proteins were evaluated by Western blot analysis. Vascular endothelial growth factor (VEGF) level was investigated by immunohistochemical staining. Enzyme-linked immunosorbent serologic assay was applied to detect the secretion of inflammatory cytokines. Furthermore, hematoxylin and eosin staining was applied to observe tissue injuries. RESULTS: Ag85B affected WI-38 cell viability in a limited manner, while TA notably suppressed Ag85B-treated WI-38 cell viability. TA induced the apoptosis of Ag85B-treated WI-38 cells in a dose-dependent manner. In addition, Ag85B-treated WI-38 cells demonstrated the upregulation of interleukin (IL)-6, tumor necrosis factor-α (TNF-α), interferon gamma (IFN-γ), and fibrotic proteins (transforming growth factor-beta [TGF-ß] and vascular endothelial growth factor [VEGF]), which can be significantly destroyed by the TA. Meanwhile, TA reversed Ag85-induced inhibition of cell autophagy by mediation of p62, LC3, and Beclin1. Furthermore, silencing of SIRT1/FOXO3a pathway could reverse the effect of TA on the autophagy of Ag85B-treated cells. CONCLUSION: TA significantly induced the autophagy of fibroblasts in Ag85B-treated cells by mediation of SIRT1/FOXO3 pathway. This study established a new theoretical basis for exploring strategies against TSTB.

Bronchoscopic instillation of amphotericin B is a safe and effective measure to treat pulmonary mycosis.

Background and objectives: In recent years, there has been a significant increase in the prevalence of pulmonary mycosis disease, and its mortality has increased. There are very few studies on treating pulmonary mycosiss with bronchoscopic instillation of amphotericin B. This study investigated the clinical efficacy and safety of bronchoscopic instillation of amphotericin B for treating pulmonary mycosiss. Methods: This was a multi-centre, retrospective clinical study of 80 patients with pulmonary mycosiss who were treated with bronchoscopic instillation of amphotericin B. The efficacy and safety of this treatment were evaluated. Results: Eighty patients were included {51 males; mean [standard deviation (SD)] age, 46 (15.9) years}. The most common underlying cause was haematological malignancy (73.75%). The mean number of bronchoscopic instillations of amphotericin B was 2.4 (SD 1.5). In terms of treatment success, 58 (72.5%) patients achieved complete or partial changes on imaging after treatment. A total of 62 (77.5%) patients achieved complete or partial changes on imaging and/or local limitation of the mycosis infection. Seventy-six (95%) patients achieved complete or partial changes on imaging and/or local limitation of mycosis infection and/or an immunotherapy time window. The efficacy rates for treatment of Aspergillus and Mucor infections in terms of the three treatment success criteria described above were 73.81% vs. 63.64%, 80.95% vs. 72.73%, and 92.86% vs. 90.91%, respectively. Conclusion: Bronchoscopic instillation of amphotericin B is safe and effective for treatment of pulmonary mycosiss.

[Management of benign tracheal stenosis by intubation dilatation under flexible bronchoscopic guidance].

OBJECTIVE: To evaluate the efficacy and safety of intubation dilatation under flexible bronchoscopic guidance in the management of benign tracheal stenosis. METHODS: A retrospective analysis of the clinical data was performed for 12 patients with benign tracheal stenosis from March 2010 to August 2011. There were 5 males and 7 females with a mean age of 37 ± 11 years old (range: 27 - 65). They were treated by intubation dilatation with different sizes under bronchoscopic guidance. And balloon dilatation was also performed for left or right main stem bronchial stenosis. And metal stents were implanted if necessary. Airway diameter, dyspnea index, complications and blood gas analysis were evaluated in all patients. And the forced expiratory volume in one second (FEV(1)) was tested in 9 cases before and after the treatments of intubation dilation, balloon dilation and other interventions. RESULTS: One to five attempts of intubation dilation were required to achieve satisfactory dilatation. There was immediate postoperative relief of dyspnea for all 12 cases. And PaO2 and SaO2 rose markedly, but PaCO2 declined after intervention. The effective rate of intubation dilation was 100%. The average airway diameter increased from (5.7 ± 1.2) to (12.2 ± 2.1) mm and FEV(1) improved from (0.67 ± 0.13) to (1.73 ± 0.37) L (P < 0.01). CONCLUSION: The minimally invasive management of benign tracheal stenosis with intubation dilatation is both safe and efficacious.

Knockdown of long non­coding RNA DLEU2 suppresses idiopathic pulmonary fibrosis by regulating the microRNA­369­3p/TRIM2 axis.

Idiopathic pulmonary fibrosis (IPF) is the most common form of idiopathic interstitial pneumonia with an increasing incidence. In the present study, Genome Expression Omnibus (GEO) datasets (GSE10667, GSE24206 and GSE32537) were applied to identify lncRNA DLEU2 in IPF. Through prediction using starBase, TargetScan, miRTarBase and miRDB, tripartite motif containing 2 (TRIM2) and prostaglandin F2 receptor inhibitor (PTGFRN) were found to be upregulated in IPF. DLEU2 expression, the mRNA expression of TRIM2 and PTGFRN, and miR­369­3p expression in A549 cells and lung tissues were detected by RT­qPCR. The protein expression of TRIM2 and PTGFRN in lung tissues and A549 cells was detected by western blot analysis. The proliferation and migration of A549 cells was respectively detected by CCK­8 assay and wound healing assay. The expression of collagen I, α­smooth muscle actin (SMA) and E­cadherin was detected by immunofluorescence assay in A549 cells, and collagen I expression was detected by immunohistochemistry assay in lung tissues. The expression of collagen I, α­SMA and E­cadherin was also detected by western blot analysis in A549 cells and lung tissues. Dual­luciferase reporter assay was used to confirm the association between DLEU2 and miR­369­3p, and miR­369­3p and TRIM2. As a result, DLEU2 expression was found to be upregulated in IPF and in transforming growth factor (TGF)­ß1­stimulated A549 cells. The silencing of DLEU2 inhibited the TGF­ß1­induced proliferation, migration and epithelial­mesenchymal transition (EMT) of A549 cells and bleomycin (BLM)­induced pulmonary fibrosis in mice. TRIM2 expression was increased and miR­369­3p expression was decreased in the lung tissues of mice with BLM­induced fibrosis and in TGF­ß1­stimulated A549 cells. DLEU2 directly targeted miR­369­3p. The effect of the silencing of DLEU2 on TGF­ß1­stimulated A549 cells was suppressed by the silencing of miR­369­3p. TRIM2 was the target protein of miR­369­3p. On the whole, the present study demonstrates that the silencing of DLEU2 suppressed IPF by upregulating miR­369­3p expression and downregulating TRIM2 expression.

Anti-fibrosis activity of quercetin attenuates rabbit tracheal stenosis via the TGF-ß/AKT/mTOR signaling pathway.

AIMS: This study aimed to explore the possible mechanism of trauma-induced laryngotracheal stenosis and potential protective and therapeutic efficacy of quercetin on trauma-induced laryngotracheal stenosis. MAIN METHODS: The expression and activity of fibrotic factors [interleukin (IL)-6, IL-8, autophagy related 5 (ATG5), collagen (COL)-1, tumor growth factor (TGF)-ß COL-3, microtubule-associated proteins 1A/1B light chain 3A (LC3), and vascular endothelial growth factor (VEGF)] and fibrotic signaling mediators [mammalian target of rapamycin (mTOR) and phosphorylated AKT (pAKT)] were detected by real-time quantitative PCR (qRT-PCR), ELISA, Western blot, and immunohistochemical staining, respectively, in the lipopolysaccharide (LPS)-induced WI-38 (a human embryonic lung fibroblast cell line) cellular fibrotic model and a trauma-induced rabbit tracheal stenosis model, with and without quercetin treatment. KEY FINDINGS: Pre-treatment with quercetin significantly reversed the LPS-induced upregulation of pro-fibrotic factors (IL-6, IL-8, COL-1, COL-3, LC3) and fibrotic signaling mediators (mTOR and AKT), and it induced the downregulation of ATG5 in the WI-38 cells. Furthermore, the anti-fibrotic activity of quercetin was confirmed in the trauma-induced rabbit tracheal stenosis model. Thus, the nasogastric administration of quercetin attenuated the tracheal stenosis of the rabbit tracheal stenosis model, in addition to effectively reversing an increase in pro-fibrotic factors (VEGF, IL-6, TGF-ß, COL-1, and COL-3) and fibrotic signaling mediators (mTOR and AKT), as well as downregulating ATG5 of the rabbit tracheal stenosis model. SIGNIFICANCE: Quercetin exhibits anti-fibrotic activity by inhibiting pro-fibrotic factors and AKT/mTOR signaling pathway, in addition to activating autophagy activity. This study provided experimental evidence supporting the application of quercetin in tracheal stenosis, clinically.