scholarly journals Mechanism underlying the role of Miaoyao Fanggan sachet in up-regulating respiratory tract immunity through TLR-MyD88-NFκB signaling pathway

2020 ◽  
Author(s):  
Mingjie Zhu ◽  
Guoquan Jia ◽  
Quan Zhang ◽  
Guojun Li ◽  
Hong Li ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Intervention Group ◽  
Immunofluorescent Staining ◽  
Lung Injury Score ◽  
Normal Lung ◽  
Mouse Lung ◽  
Epithelial Cell Line ◽  
Lung Epithelial ◽  
Liver And Kidney

Abstract Background To elucidate the mechanism underlying the role of Miaoyao Fanggan sachet (MFS) in improving respiratory immunity through the TLR-MyD88-NFκB signaling pathway in MyD88 knockout mouse models and human normal lung epithelial cell line (BEAS-2B).Methods Sixteen male C57BL/6J mice (SPF grade) were randomly divided into the control (n = 8) and MFS groups (n = 8) and 16 male MyD88 knockout mice were randomly assigned into the MyD88 knockout (MyD88-KO group, n = 8) and MyD88 KO + MFS groups (n = 8). Mice in the MFS intervention group were continuously treated with MFS inhalation from 8:00 am to 8:00 pm for 30 d and the sachets were replaced every 6 d. Hematoxylin-eosin (H.E.) staining was used to observe the pathological changes of lung tissues. The expression levels of TAK1, NFκB p65, IκB and IL6 in mouse lung tissues were detected by immunohistochemical staining, immunofluorescent staining, Western blot and qRT-PCR, respectively.Results Mikawa’s lung injury score did not significantly differ between the control and MFS groups (both P > 0.05). The parameters related to liver and kidney function did not significantly differ among four groups (all P > 0.05). The TAK1, NFκB p65, IκB and IL6 mRNA and protein in the MFS group were significantly up-regulated than those in the other groups (all P < 0.05). Compared with the MyD88-KO group, they were significantly up-regulated in the MyD88-KO + MFS group (P < 0.05).Conclusion HFS can improve the respiratory immunity probably by up-regulating the expression level of MyD88, activating the TLR-MyD88-NFκB signaling pathway and stimulating the release of cytokines in the downstream pathway.

scholarly journals The Role of LncRNA H19 in MAPK Signaling Pathway Implicated in the Progression of Bronchopulmonary Dysplasia

2020 ◽  
Vol 29 ◽  
pp. 096368972091829 ◽  
Author(s):  
Wenhui Mo ◽  
Yi Li ◽  
Weijie Chang ◽  
Yaoming Luo ◽  
Bingbin Mai ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Bronchopulmonary Dysplasia ◽  
Respiratory Illness ◽  
Mapk Signaling ◽  
Mapk Signaling Pathway ◽  
Mitogen Activated Protein Kinase ◽  
Inflammatory Factors ◽  
Normal Lung ◽  
Mouse Lung

Bronchopulmonary dysplasia (BPD), also known as neonatal chronic lung disease, is an important cause of respiratory illness in preterm newborns that results in significant morbidity and mortality. Long noncoding RNAs (lncRNAs) have been discovered with many biological functions. However, the role of lncRNAs in the pathogenesis of BPD remains poorly understood. Here, we established a mouse lung injury model that mimicked human BPD. Subsequently, we found the lncRNA H19 expression level was significantly increased in BPD compared with normal lung tissues using quantitative real-time polymerase chain reaction. Next, we observed that overexpression of lncRNA H19 enhanced mitogen-activated protein kinase (MAPK) signaling pathway. In addition, we also found that dysfunction of lncRNA H19 altered the expression of inflammatory factors. Thus, our study validates that lncRNA H19 contributes to the progression of BPD by regulating MAPK signaling pathway, which could be used as a potential target for treating BPD.

Mechanism of Tripchlorolide Inhibiting Hedgehog Signaling Pathway to Delay Lung Fibrosis

2020 ◽  
Vol 10 (7) ◽  
pp. 992-998
Author(s):  
Shirui Tan ◽  
Qingrong Li ◽  
Feifei Duan ◽  
Qingqing Yuan ◽  
Gang Deng
Keyword(s):  
Signaling Pathway ◽  
Lung Tissue ◽  
Hedgehog Signaling ◽  
Tissue Injury ◽  
Collagen I ◽  
Lung Fibroblasts ◽  
Lung Injury Score ◽  
Mouse Lung ◽  
Hedgehog Signaling Pathway ◽  
Gli 1

Background: The paper aimed to determine whether Tripchlorolide can exert anti-fibrotic effects by up-regulating Sufu to influence the Hedgehog signaling pathway, and to promote the application of Tripchlorolide in diseases associated with the Hedgehog signaling pathway. Material and Methods: In vivo experiment, bronchoalveolar lavage was performed on the 7th day after intratracheal instillation of bleomycin. The number of inflammatory cells and IL-6 levels were analyzed to observe the anti-inflammatory effect of Tripchlorolide. The lung tissues were obtained on the 14th day. The lung tissue injury was observed, and the expressions of a-SMA, collagen I, TGF-β i, Gli 1 and Sufu were determined. In vitro experiments, mouse lung was transfected with lentivirus to make fibroblasts. The Sufii was overexpressed, the changes in Gli 1 and Gli 2 levels were observed. The effects of the overexpressed Sufti of on the phenotype transformed a-SMA and collagen I levels in TGF-β 1 induced mouse lung fibroblasts were observed. Results: After administration with bleomycin, IL-6 level, total number of cells, neutrophils and macrophages in BALF of the Tripchlorolide group mice were reduced. The lung injury score, the expressions of lung tissue TGF-βl, a-SMA and Collagen I were low. The expression of Sufu was high, while the expression of Gli 1 was low. Compared with the groups in which cells were not transfected with lentivirus, the expression of Sufu in the transfected cells increased, and the expressions of Gli 1 and Gli 2 decreased. After stimulation with TGF-β Sufu decreased. Conclusion: Tripchlorolide can inhibit Hedgehog signaling pathway by up-regulating Sufu to exert an anti-fibrotic effect.

scholarly journals Suppression of TLR4-MyD88 signaling pathway attenuated chronic mechanical pain in a rat model of endometriosis

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Wenliang Su ◽  
Huan Cui ◽  
Danning Wu ◽  
Jiawen Yu ◽  
Lulu Ma ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
High Mobility ◽  
Adaptor Protein ◽  
Immunofluorescent Staining ◽  
Inhibitory Peptide ◽  
Tlr4 Signaling ◽  
Innate Immunity Pathway ◽  
Myd88 Signaling ◽  
Myeloid Differentiation Factor

Abstract Background As a classic innate immunity pathway, Toll-like receptor 4 (TLR4) signaling has been intensively investigated for its function of pathogen recognition. The receptor is located not only on immune cells but also on sensory neurons and spinal glia. Recent studies revealed the involvement of neuronal TLR4 in different types of pain. However, the specific role of TLR4 signaling in the pain symptom of endometriosis (EM) remains obscure. Methods The rat endometriosis model was established by transplanting uterine horn tissue into gastrocnemius. Western blotting and/or immunofluorescent staining were applied to detect high mobility group box 1 (HMGB1), TLR4, myeloid differentiation factor-88 adaptor protein (MyD88), and nuclear factor kappa-B-p65 (NF-κB-p65) expression, as well as the activation of astrocyte and microglia. The antagonist of TLR4 (LPS-RS-Ultra, LRU) and MyD88 homodimerization inhibitory peptide (MIP) were intrathecally administrated to assess the behavioral effects of blocking TLR4 signaling on endometriosis-related pain. Results Mechanical hyperalgesia was observed at the graft site, while HMGB1 was upregulated in the implanted uterine tissue, dorsal root ganglion (DRG), and spinal dorsal horn (SDH). Compared with sham group, upregulated TLR4, MyD88, and phosphorylated NF-κB-p65 were detected in the DRG and SDH in EM rats. The activation of astrocytes and microglia in the SDH was also confirmed in EM rats. Intrathecal application of LRU and MIP alleviated mechanical pain on the graft site of EM rats, with decreased phosphorylation of NF-κB-p65 in the DRG and reduced activation of glia in the SDH. Conclusions HMGB1-TLR4-MyD88 signaling pathway in the DRG and SDH may involve in endometriosis-related hyperpathia. Blockade of TLR4 and MyD88 might serve as a potential treatment for pain in endometriosis.

scholarly journals Dynamic Observation of Autophagy and Transcriptome Profiles in a Mouse Model of Bleomycin-Induced Pulmonary Fibrosis

2021 ◽  
Vol 8 ◽  
Author(s):  
Yani Wang ◽  
Siqi Hu ◽  
Lisha Shen ◽  
Song Liu ◽  
Linyan Wan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Pulmonary Fibrosis ◽  
Mouse Model ◽  
Muscle Contraction ◽  
Signaling Pathway ◽  
Lung Tissue ◽  
Critical Role ◽  
Mouse Lung ◽  
Mouse Lung Tissue

Pulmonary fibrosis is a group of progressive, fibrotic, and fatal lung diseases, and the role of autophagy in pulmonary fibrosis is controversial. In the current research, we dynamically observed a bleomycin-induced pulmonary fibrosis mouse model after 3, 7, 14, 21, and 28 days and investigated the expression of autophagy markers. We found that autophagy markers were not significantly changed on the indicated days in the mouse lung tissue. Then, RNA-Seq was used to analyze the gene expression and associated functions and pathways in fibrotic lung tissue on different days post-bleomycin. In addition, short time series expression miner (STEM) analysis was performed to explore the temporal post-bleomycin gene expression. Through STEM, continually up- or downregulated profiles did not demonstrate the critical role of autophagy in the development of fibrosis. Furthermore, gene ontology (GO) annotations showed that continually upregulated profiles were mainly related to fibrosis synthesis, extracellular space, and inflammation, while enriched pathways were mainly related to the PI3K-Akt signaling pathway, ECM–receptor interactions, and focal adhesion signaling pathway. For continually downregulated profiles, GO annotations mainly involved sarcomere organization, muscle contraction, and muscle fiber development. The enriched KEGG signaling pathways were the cAMP signaling pathway, cGMP-PKG signaling pathway, calcium signaling pathway, and cardiac muscle contraction. Moreover, we analyzed autophagy-related genes’ expression in specific cells from a publicly available database of three human and one animal study of pulmonary fibrosis using single-cell sequencing technology. All results consistently demonstrated no critical role of autophagy in the pathogenesis of pulmonary fibrosis. In summary, autophagy may not critically and consistently change during the development of pulmonary fibrosis at different stages post-bleomycin in a mouse model. These continually up- or downregulated profiles, including gene profiles, and the corresponding functions and pathways may provide mechanistic insights into IPF therapy.

scholarly journals Francisella tularensis Invasion of Lung Epithelial Cells

2008 ◽  
Vol 76 (7) ◽  
pp. 2833-2842 ◽  
Author(s):  
Robin R. Craven ◽  
Joshua D. Hall ◽  
James R. Fuller ◽  
Sharon Taft-Benz ◽  
Thomas H. Kawula
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Francisella Tularensis ◽  
Bacterial Pathogen ◽  
Alveolar Epithelial Cells ◽  
Lung Epithelial Cells ◽  
Mouse Lung ◽  
Epithelial Cell Line ◽  
Bacterial Surface ◽  
Lung Epithelial

ABSTRACT Francisella tularensis, a gram-negative facultative intracellular bacterial pathogen, causes disseminating infections in humans and other mammalian hosts. Macrophages and other monocytes have long been considered the primary site of F. tularensis replication in infected animals. However, recently it was reported that F. tularensis also invades and replicates within alveolar epithelial cells following inhalation in a mouse model of tularemia. TC-1 cells, a mouse lung epithelial cell line, were used to study the process of F. tularensis invasion and intracellular trafficking within nonphagocytic cells. Live and paraformaldehyde-fixed F. tularensis live vaccine strain organisms associated with, and were internalized by, TC-1 cells at similar frequencies and with indistinguishable differences in kinetics. Inhibitors of microfilament and microtubule activity resulted in significantly decreased F. tularensis invasion, as did inhibitors of phosphatidylinositol 3-kinase and tyrosine kinase activity. Collectively, these results suggest that F. tularensis epithelial cell invasion is mediated by a preformed ligand on the bacterial surface and driven entirely by host cell processes. Once internalized, F. tularensis-containing endosomes associated with early endosome antigen 1 (EEA1) followed by lysosome-associated membrane protein 1 (LAMP-1), with peak coassociation frequencies occurring at 30 and 120 min postinoculation, respectively. By 2 h postinoculation, 70.0% (± 5.5%) of intracellular bacteria were accessible to antibody delivered to the cytoplasm, indicating vacuolar breakdown and escape into the cytoplasm.

scholarly journals Sp3 regulates Fas expression in lung epithelial cells

1998 ◽  
Vol 333 (1) ◽  
pp. 209-213 ◽  
Author(s):  
Huiling PANG ◽  
Kathleen MIRANDA ◽  
Alan FINE
Keyword(s):  
Epithelial Cells ◽  
Promoter Activity ◽  
Functional Results ◽  
Lung Epithelial Cells ◽  
Site Directed Mutagenesis ◽  
Mouse Lung ◽  
Epithelial Cell Line ◽  
Upstream Sequence ◽  
Mobility Shift ◽  
Lung Epithelial

By transducing an apoptotic signal in immune effector cells, Fas has been directly implicated in the control of immunological activity. Expression and functional results, however, have also suggested a role for Fas in regulating cell turnover in specific epithelial populations. To characterize factors responsible for Fas expression in epithelial cells, approximately 3 kb of the 5´ flanking region of the mouse Fas gene was isolated. By rapid amplification of cDNA ends and primer extension, transcriptional start sites were identified within 50 bp upstream of the translation start site. Transient transfection of promoter–luciferase constructs in a mouse lung epithelial cell line, MLE-15, localized promoter activity to the first 77 bp of upstream sequence. By using a 60 bp DNA probe (-18 to -77) in electrophoretic mobility-shift assays, three shifted complexes were found. Incubation with excess cold Sp1 oligonucleotide or an anti-Sp3 antibody inhibited complex formation. Site-directed mutagenesis of the Sp1 site resulted in 60–70% loss of promoter activity. In Drosophila SL-2 cells, promoter activity was markedly increased by co-transfection of an Sp3 expression construct. These results show that the Sp3 protein is involved in regulating Fas gene expression in lung epithelial cells.

scholarly journals Retinoic acid and dexamethasone affect RAR-β and surfactant protein C mRNA in the MLE lung cell line

1998 ◽  
Vol 274 (1) ◽  
pp. L1-L7 ◽  
Author(s):  
Mary A. Grummer ◽  
Richard D. Zachman
Keyword(s):  
Retinoic Acid ◽  
Cell Line ◽  
Lung Development ◽  
Actinomycin D ◽  
Surfactant Protein ◽  
Mouse Lung ◽  
Epithelial Cell Line ◽  
Surfactant Protein C ◽  
Lung Epithelial ◽  
Lung Cell Line

Lung development and surfactant biosynthesis are affected by retinoic acid (RA) and dexamethasone (Dex). Using a mouse lung epithelial cell line, we are exploring RA-Dex interactions through the study of RA and Dex effects on RA receptor (RAR) and surfactant protein (SP) C mRNA expression. RA increased expression of RAR-β (5.5 times) and SP-C (2 times) mRNA, with maximal effects at 24 h and at 10−6 M. The RA induction was not inhibited by cycloheximide, suggesting RA affects transcription. With added actinomycin D, RA did not affect the disappearance rate of RAR-β mRNA, but SP-C mRNA degradation was slowed, indicating an effect on SP-C mRNA stability. Dex decreased RAR-β and SP-C expression to 75 and 70% of control values, respectively, with greatest effects at 48 h and at 10−7 M. There was no effect of Dex on either RAR-β or SP-C mRNA disappearance with actinomycin D. However, cycloheximide prevented the effect of Dex. Despite Dex, RA increased both RAR-β and SP-C mRNA. This work suggests that RA and Dex affect RAR-β and SP-C genes by different mechanisms.

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