scholarly journals A Chinese Traditional Therapy for Bleomycin-Induced Pulmonary Fibrosis in Mice

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
Lifang Sun ◽  
Minjie Mao ◽  
Zhisheng Yan ◽  
Cuiyun Zuo ◽  
Xiaojie Zhang
Keyword(s):  
Nitric Oxide ◽  
Dna Damage ◽  
Pulmonary Fibrosis ◽  
Smooth Muscle Actin ◽  
Tissue Growth ◽  
Alpha Smooth Muscle Actin ◽  
Real Time Quantitative Pcr ◽  
Incidence And Mortality ◽  
Total Antioxidant ◽  
Scge Assay

Pulmonary fibrosis is a chronic and fatal disease of lung tissue with high incidence and mortality in the world. The exploration of effective treatment for pulmonary fibrosis remains an urgent challenge. In our study, Qingfei Xieding was investigated as a novel Chinese traditional patent medicine against pulmonary fibrosis. A pulmonary fibrosis mouse model was constructed by injecting with bleomycin sulfate. Following Qingfei Xieding administration, lung samples were collected to assess pulmonary phenotype changes by analyzing lung coefficient, wet/dry, and histopathologic section. Levels of nitric oxide (NO), hydroxyproline (HYP), malondialdehyde (MDA), and total antioxidant capacity were measured to evaluate the degree of oxidation. A single-cell gel electrophoresis (SCGE) assay was used to evaluate bleomycin-induced DNA damage. Western blotting and real-time quantitative PCR were performed to determine the abundance of inducible nitric oxide synthase (iNOS), connective tissue growth factor (CTGF), alpha smooth muscle actin (α-SMA), and fibronectin (FN). In the present study, Qingfei Xieding administration significantly attenuated bleomycin-induced pulmonary fibrosis in mice by reducing lung coefficient, wet/dry, NO, HYP, and MDA as well as the expression of iNOS, CTGF, α-SMA, FN, and DNA damage. The results indicated that Qingfei Xieding is effective to resist oxidative damage and histopathologic lesion, serving a protection role on bleomycin-induced pulmonary fibrosis.

scholarly journals Regulation of Cellular Senescence Is Independent from Profibrotic Fibroblast-Deposited ECM

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1628
Author(s):  
Kaj E. C. Blokland ◽  
Habibie Habibie ◽  
Theo Borghuis ◽  
Greta J. Teitsma ◽  
Michael Schuliga ◽  
...  
Keyword(s):  
Growth Factor ◽  
Cellular Senescence ◽  
Transforming Growth Factor ◽  
Cell Function ◽  
Smooth Muscle Actin ◽  
Transforming Growth Factor Β1 ◽  
Tissue Growth ◽  
Alveolar Epithelial Cells ◽  
Lung Fibroblasts ◽  
Alpha Smooth Muscle Actin

Idiopathic pulmonary fibrosis (IPF) is a devastating lung disease with poor survival. Age is a major risk factor, and both alveolar epithelial cells and lung fibroblasts in this disease exhibit features of cellular senescence, a hallmark of ageing. Accumulation of fibrotic extracellular matrix (ECM) is a core feature of IPF and is likely to affect cell function. We hypothesize that aberrant ECM deposition augments fibroblast senescence, creating a perpetuating cycle favouring disease progression. In this study, primary lung fibroblasts were cultured on control and IPF-derived ECM from fibroblasts pretreated with or without profibrotic and prosenescent stimuli, and markers of senescence, fibrosis-associated gene expression and secretion of cytokines were measured. Untreated ECM derived from control or IPF fibroblasts had no effect on the main marker of senescence p16Ink4a and p21Waf1/Cip1. However, the expression of alpha smooth muscle actin (ACTA2) and proteoglycan decorin (DCN) increased in response to IPF-derived ECM. Production of the proinflammatory cytokines C-X-C Motif Chemokine Ligand 8 (CXCL8) by lung fibroblasts was upregulated in response to senescent and profibrotic-derived ECM. Finally, the profibrotic cytokines transforming growth factor β1 (TGF-β1) and connective tissue growth factor (CTGF) were upregulated in response to both senescent- and profibrotic-derived ECM. In summary, ECM deposited by IPF fibroblasts does not induce cellular senescence, while there is upregulation of proinflammatory and profibrotic cytokines and differentiation into a myofibroblast phenotype in response to senescent- and profibrotic-derived ECM, which may contribute to progression of fibrosis in IPF.

scholarly journals Changes in histological structure and nitric oxide synthase expression in aorta of rats supplemented with bee pollen or whey protein

2019 ◽  
Vol 44 (11) ◽  
pp. 1150-1158
Author(s):  
Michał K. Zarobkiewicz ◽  
Mirosław A. Sławiński ◽  
Ewelina Wawryk-Gawda ◽  
Mateusz M. Woźniakowski ◽  
Emilia Kulak-Janczy ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Whey Protein ◽  
Smooth Muscle Actin ◽  
Histological Evaluation ◽  
Moderate Physical Activity ◽  
Histological Structure ◽  
Alpha Smooth Muscle Actin ◽  
Bee Pollen ◽  
Oxide Synthase

Various protein-based supplements are at least periodically consumed by 30%–40% of sportspeople. The current study compares cardiovascular effects of diet supplementation with 2 different protein-rich products: bee pollen and whey protein. Thirty Wistar rats were divided into 2 groups, one subjected to daily moderate physical activity and one not. Each group consisted of 3 subgroups: control, whey-protein-supplemented, and bee-pollen-supplemented. After 8 weeks, rats were decapitated, and proximal parts of thoracic aortas were collected and embedded in paraffin blocks. Histological slides were stained according to standard hematoxylin and eosin, Masson’s trichrome, and Verhoeff – Van Gieson staining. Special immunohistochemical stains against neuronal nitric oxide synthase (nNOS), endothelial nitric oxide synthase (eNOS), and alpha smooth muscle actin were also prepared. Histological evaluation revealed noticeable changes in all supplemented groups: disturbances in elastic laminae, slight increase in collagen deposition, and significantly lowered nNOS and eNOS expression. The prevalence of small atherosclerotic plaques was the highest in non-running supplemented groups, while in running supplemented groups it resembled the prevalence in control groups. Both running groups had thinner tunica media than control. Both supplements exert visible effects on aortic structure, but the difference between them is far less evident. In some aspects, however, the bee pollen seems to be even slightly more harmful, which may be related to various possible contaminants like mycotoxins or pesticides.

scholarly journals Danggui Buxue Tang Ameliorates Bleomycin-Induced Pulmonary Fibrosis by Suppressing the TLR4/NLRP3 Signaling Pathway in Rats

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Jiepeng Wang ◽  
Hao Wang ◽  
Fang Fang ◽  
Chaoyi Fang ◽  
Shaoxian Wang ◽  
...  
Keyword(s):  
Pulmonary Fibrosis ◽  
Signaling Pathway ◽  
Smooth Muscle Actin ◽  
Interleukin 1 ◽  
Sprague Dawley ◽  
Alpha Smooth Muscle Actin ◽  
Potent Effect ◽  
Necrosis Factor Alpha ◽  
Danggui Buxue Tang ◽  
Collagen Iii

Objective. To investigate the effects of Danggui Buxue Tang (DBT) on rats with pulmonary fibrosis (PF) and the underlying mechanism. Methods. Sixty specific pathogen-free (SPF) male Sprague-Dawley (SD) rats were randomly divided into 4 groups: control, PF, prednisone treatment, and DBT treatment. Intratracheal instillation of bleomycin (BLM) was performed to establish a PF rat model. DBT was administered to PF rats concurrently for 2 weeks. Lung samples were then collected for HE and Masson staining after pulmonary function testing, and semiquantitative analysis for the degree of alveolitis and fibrosis was performed using the Szapiel and Ashcroft score systems. Myeloperoxidase (MPO) activity, hydroxyproline (HYP), hyaluronic acid (HA), and inflammatory cytokine content were measured. Western blotting was performed to detect fibrotic marker and TLR4/NLRP3 signaling pathway changes. Results. Oral administration of DBT attenuated weight loss, survival rate, and pulmonary index. Lung histopathologic lesions were also reduced. DBT inhibited PF by decreasing the secretion of inflammatory cytokines and collagen deposition. Specifically, DBT reduced tumor necrosis factor-alpha (TNF-α), interleukin 1 beta (IL-1β), IL-6, HYP, alpha-smooth muscle actin (α-SMA), collagen I, and collagen III levels. Corollary experiments identified a potential mechanism involving suppression of TLR4/MyD88/NF-κB signaling pathway activation and the NLRP3/ASC/caspase-1 axis, the downstream regulatory pathway. Conclusion. DBT exhibited a potent effect on BLM-induced PF rats by inhibiting the TLR4/NLRP3 signaling pathway. Thus, DBT alleviates pulmonary inflammation to inhibit fibrotic pathology and should be considered as a candidate for the clinical treatment of PF.

Ticagrelor Ameliorates Bleomycin-Induced Pulmonary Fbrosis in Rats by Inhibition of TGF-β1/Smad3 and PI3K/AKT/mTOR Pathways

2021 ◽  
Vol 14 ◽  
Author(s):  
Hanaa Wanas ◽  
Zeinab El Shereef ◽  
Laila Rashed ◽  
Basma Emad Aboulhoda
Keyword(s):  
Pulmonary Fibrosis ◽  
Transforming Growth Factor ◽  
Epithelial Mesenchymal Transition ◽  
Therapeutic Potential ◽  
Smooth Muscle Actin ◽  
Alpha Smooth Muscle Actin ◽  
P2y12 Inhibitor ◽  
Mesenchymal Transition ◽  
Serious Disease ◽  
Tgf Β1

Background: Idiopathic pulmonary fibrosis (IPF) is a serious disease with high mortality rate. Activation of transforming growth factor (TGF)-β1 production and signalling is considered the corner stone in the epithelial-mesenchymal transition (EMT) process. EMT plays a central role in development of fibrosis in many organs including the lungs. Activated platelets is an important source of TGF-β1 and play a pivotal role in EMT and fibrosis process. The antiplatelet, ticagrelor was previously found to inhibit the EMT in different types of cancer cells, but its ability to serve as an anti-pulmonary fibrosis (PF) agent was not previously investigated. Objective: In this study, we aim to investigate the potential ability of ticagrelor to ameliorate bleomycin-induced fibrosis in rats. Methods: PF was induced in rats by intratracheal BLM at a dose of 3 mg/kg. The effect of daily daily 20 mg/kg oral ticagrelor on different histological and biochemical parameters of fibrosis was investigated. Results: Our results revealed that ticagrelor can alleviate lung fibrosis. We found that ticagrelor inhibited TGF-β1 production and suppressed Smad3 activation and signaling pathway with subsequent inhibition of Slug and Snail. In addition, ticagrelor antagonized PI3K/AKT/mTOR pathway signaling. Moreover, ticagrelor inhibited the EMT that revealed by its ability to up-regulate the epithelial markers as E-cadherin (E-cad) and to decrease the expression of the mesenchymal markers as vimentin (VIM) and alpha-smooth muscle actin (α-SMA). Conclusion: Our results suggest that the P2Y12 inhibitor, ticagrelor may have a therapeutic potential in reducing the progression of PF.

Abstract P195: Prorenin Receptor Activation Increases Profibrotic Markers and Fibroblast Like Phenotype Through MAPK-dependent ROS Formation in Mouse Renal Collecting Duct Cells

Hypertension ◽  
2016 ◽  
Vol 68 (suppl_1) ◽  
Author(s):  
Alexis A Gonzalez ◽  
Cristian Reyes-Martinez ◽  
Leonardo Zamora ◽  
Minolfa C Prieto
Keyword(s):  
Mapk Pathway ◽  
Collecting Duct ◽  
Smooth Muscle Actin ◽  
Receptor Activation ◽  
Tissue Growth ◽  
Tubular Damage ◽  
Plasminogen Activator Inhibitor 1 ◽  
Alpha Smooth Muscle Actin ◽  
Prorenin Receptor ◽  
Ros Formation

Binding of prorenin or renin to (pro)renin receptor (PRR) activates mitogen activated protein kinases (MAPK)/extracellular regulated kinases 1/2 (ERK 1/2), which have been proposed as mediators in tissue damage. Prorenin, renin, and PRR are overexpressed in the collecting duct (CD) in diabetes and hypertension. Activation of the PRR upregulates profibrotic markers through reactive oxygen species (ROS) formation; however, the exact mechanisms have not been established. We hypothesized that the activation of PRR increases the expression of profibrotic markers through MAPK-dependent ROS formation in CD cells. To address this hypothesis, a mouse renal CD cell line (M-1) was treated with recombinant prorenin plus ROS or MAPK inhibitor to evaluate their effect on the expression of profibrotic markers. Recombinant prorenin increased ROS formation and expression of alpha smooth muscle actin (α-SMA), connecting tissue growth factor (CTGF) and plasminogen activator inhibitor-1 (PAI-1). Induction of profibrotic factors was blunted by antioxidant p-coumaric acid, and partially prevented by lyophilic antioxidant trolox C or ascorbic acid. Inhibition of MAPK pathway (PD98059) prevented the prorenin-dependent ROS formation and augmentation of profibrotic factors, as well as fibroblast-like phenotype. Knockdown of PRR did not show effects on cell viability in M-1 cells and partially reduced the augmentation of fibronectin, collagen I and fibroblast-like phenotype induced by prorenin treatment. These results suggest that the activation of CD-derived PRR plays a role in the development of tubular damage.

The Strain Response of Lung Myofibroblasts Cultured on Electrospun Polycaprolactone Nanofibers

2013 ◽  
Author(s):  
Timothy J. Fee ◽  
Yong Zhou ◽  
Lauren E. Marshall ◽  
Joel L. Berry
Keyword(s):  
Smooth Muscle ◽  
Idiopathic Pulmonary Fibrosis ◽  
Pulmonary Fibrosis ◽  
Smooth Muscle Actin ◽  
Strain Response ◽  
Muscle Actin ◽  
Alpha Smooth Muscle Actin

Idiopathic Pulmonary Fibrosis is a devastating condition characterized by excessive localized production of collagen in the lungs. Over 131,000 people are living with IPF in America (1, 2). There is currently no known treatment or cure for the disease. It has recently been shown that IPF myofibroblasts are sensitive to the stiffness of their substrate. Specifically, alpha Smooth Muscle Actin (alpha-SMA), a known indicator of IPF activity, was differentially produced on soft vs. stiff substrates (3). This suggests a mechanotransduction pathway within the IPF myofibroblasts.

scholarly journals Akt1 regulates pulmonary fibrosis via modulating IL-13 expression in macrophages

2019 ◽  
Vol 25 (7) ◽  
pp. 451-461 ◽  
Author(s):  
Yunjuan Nie ◽  
Yudong Hu ◽  
Kaikai Yu ◽  
Dan Zhang ◽  
Yinze Shi ◽  
...  
Keyword(s):  
Idiopathic Pulmonary Fibrosis ◽  
Pulmonary Fibrosis ◽  
Smooth Muscle Actin ◽  
In Vivo Studies ◽  
Muscle Actin ◽  
Alpha Smooth Muscle Actin ◽  
Fibrosis Progression ◽  
Matrix Components ◽  
Deficient Mice

Idiopathic pulmonary fibrosis is a progressive interstitial pneumonia characterised by fibroblast accumulation, collagen deposition and extracellular matrix (ECM) remodelling. It was reported that Akt1 mediated idiopathic pulmonary fibrosis progression through regulating the apoptosis of alveolar macrophage, while its effect on macrophage-produced cytokines remains largely unknown. In the present study, we first examined the phosphorylation of Akt1 in lung sections from idiopathic pulmonary fibrosis patients by immunohistochemistry before applying a bleomycin-induced idiopathic pulmonary fibrosis model using Akt1−/− mice and Akt1+/+ littermates. The results showed that Akt1 was remarkably up-regulated in idiopathic pulmonary fibrosis patients, while in vivo studies revealed that Akt1-deficient mice had well-preserved alveolar structure and fewer collagens, secreted fewer matrix components, including alpha smooth-muscle actin and fibronectin and survived significantly longer than Akt1+/+ littermates. Additionally, the pro-fibrogenic cytokine IL-13 was down-regulated at least twofold in Akt1−/−mice compared to the Akt1+/+group on d 3 and 7 after bleomycin treatment. Furthermore, it was found that Akt1–/– macrophages displayed down-regulation of IL-13 compared to Akt1+/+ macrophages in which Akt1 was phosphorylated in response to IL-33 stimulation. These findings indicate that Akt1 modulates pulmonary fibrosis through inducing IL-13 production by macrophages, suggesting that targeting Akt1 may simultaneously block the fibrogenic processes of idiopathic pulmonary fibrosis.

scholarly journals Targeting MAP3K19 prevents human lung myofibroblast activation both in vitro and in a humanized SCID model of idiopathic pulmonary fibrosis

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Isabelle C. Jones ◽  
Milena S. Espindola ◽  
Rohan Narayanan ◽  
Ana L. Coelho ◽  
David M. Habiel ◽  
...  
Keyword(s):  
Idiopathic Pulmonary Fibrosis ◽  
Pulmonary Fibrosis ◽  
Smooth Muscle Actin ◽  
Mitogen Activated Protein Kinase ◽  
Lung Fibroblasts ◽  
Alpha Smooth Muscle Actin ◽  
Biochemical Assays ◽  
Mitogen Activated Protein ◽  
Si Rna

AbstractIdiopathic Pulmonary Fibrosis (IPF) is a disease with a devastating prognosis characterized by unrelenting lung scarring. Aberrant activation of lung fibroblasts is a key feature of this disease, yet the key pathways responsible for this are poorly understood. Mitogen-activated protein kinase, kinase, kinase- 19 (MAP3K19) was recently shown to be upregulated in IPF and this MAPK has a key role in target gene transcription in the TGF-β pathway. Herein, we further investigate the role of MAP3K19 in cultured normal and IPF fibroblasts and in a humanized SCID mouse model of IPF employing both short interfering (si) RNA and novel small-molecule inhibitors directed at this kinase. Targeting MAP3K19 had significant inhibitory effects on the expression of both alpha smooth muscle actin and extracellular matrix in cultured human IPF fibroblasts. Quantitative protein and biochemical assays, as well as histological analysis, showed that MAP3K19 was required for the development of lung fibrosis in SCID mice humanized with IPF lung fibroblasts. MAP3K19 was required for IPF myofibroblast differentiation, and targeting its activity attenuated the profibrotic activity of these cells both in vitro and in an adoptive transfer SCID model of pulmonary fibrosis.

scholarly journals RXFP1 expression is regulated by miR-144-3p in Fibroblasts from Patients with Idiopathic Pulmonary Fibrosis

2017 ◽  
Author(s):  
Harinath Bahudhanapati ◽  
Jiangning Tan ◽  
Justin A Dutta ◽  
Stephen B Strock ◽  
Yingze Zhang ◽  
...  
Keyword(s):  
Idiopathic Pulmonary Fibrosis ◽  
Pulmonary Fibrosis ◽  
Smooth Muscle Actin ◽  
Negative Regulator ◽  
Lung Fibroblasts ◽  
Luciferase Reporter ◽  
Alpha Smooth Muscle Actin ◽  
Protein Levels ◽  
Donor Lung ◽  
Luciferase Reporter Vector

ABSTRACTRelaxin has been considered as a potential therapy for patients with pulmonary fibrosis. We have previously shown, however, that a potential limitation of relaxin-based therapy for Idiopathic Pulmonary Fibrosis (IPF) is the loss of expression of the relaxin receptor Relaxin/Insulin Like Receptor 1 (RXFP1) expression in fibroblasts. The molecular mechanism for RXFP1 down-regulation in IPF patients remains unclear. To determine whether microRNAs play a role in RXFP1 gene expression, we employed a bioinformatics approach to identify microRNAs (miRs) that are predicted to target RXFP1. By in silico analysis, we identified a putative target site in the RXFP1 mRNA for the miR-144 family. We found that miR-144-3p was upregulated in IPF fibroblasts compared to donor lung fibroblast controls. Forced miR-144-3p mimic expression reduced RXFP1 mRNA and protein levels and increased expression of the myofibroblast marker alpha-smooth muscle actin (α-SMA) in donor lung fibroblasts. IPF lung fibroblasts transfected with a miR-144-3p inhibitor increased RXFP1 expression and reduced α-SMA expression. A lentiviral luciferase reporter vector carrying the WT 3’UTR of RXFP1 was repressed more in lung fibroblasts whereas vector carrying a mutated miR-144-3p binding site exhibited less sensitivity to endogenous miR-144-3p expression, suggesting that RXFP1 is a direct target of miR-144-3p. Thus, miR-144-3p is highly expressed in IPF fibroblasts and acts as a negative regulator of RXFP1 protein expression.

Biomarkers and Heterogeneous Fibroblast Phenotype Associated with Incisional Hernia

2021 ◽  
Author(s):  
Devendra K. Agrawal ◽  
Finosh G. Thankam ◽  
Nicholas K. Larsen ◽  
Ann Verghese ◽  
Thao-Nguyen Bui ◽  
...  
Keyword(s):  
Growth Factor ◽  
Connective Tissue ◽  
Incisional Hernia ◽  
Connective Tissue Growth Factor ◽  
Organ Procurement ◽  
Smooth Muscle Actin ◽  
Specific Protein ◽  
Tissue Growth ◽  
Hernia Surgery ◽  
Alpha Smooth Muscle Actin

Abstract Development of incisional hernia (IH) is multifactorial but inflammation and abdominal wall ECM (extracellular matrix) disorganization are key pathological events. We investigated if the differential expression of fibroblast biomarkers reflects the cellular milieu and the dysregulated ECM in IH tissues. Expression of fibroblast biomarkers, including connective tissue growth factor, alpha-smooth muscle actin (α-SMA), CD34 (cluster of differentiation 34), cadherin-11 and fibroblast specific protein 1 (FSP1), was examined by histology and immunofluorescence in the hernial-fascial ring/neck tissue (HRT) and hernia sack tissue (HST) harvested from the patients undergoing hernia surgery and compared with normal fascia (FT) and peritoneum (PT) harvested from brain-dead healthy subjects undergoing organ procurement for transplantation. The H&E staining revealed alterations in tissue architecture, fibroblast morphology, and ECM organization in the IH tissues compared to control. The biomarker for undifferentiated fibroblasts, CD34, was significantly higher in HST and decreased in HRT than the respective FT and PT controls. Also, the findings revealed an increased level of CTGF (connective tissue growth factor) with decrease in α-SMA in both HRT and HST compared to the controls. In addition, an increased level of FSP1 (fibroblast specific protein 1) and cadherin-11 in HRT with decreased level in HST were observed relative to the respective controls (FT and PT). Hence, these findings support the heterogeneity of fibroblast population at the laparotomy site that could contribute to the development of IH. Understanding the mechanisms causing the phenotype switch of these fibroblasts would open novel strategies to prevent the development of IH following laparotomy.

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