scholarly journals Nintedanib suppresses expression of extracellular matrix proteins in TGF-β1-stimulated primary human fibroblasts of the Tenon

2021 ◽  
Vol 7 (2) ◽  
pp. 367-370
Author(s):  
Andreas Brietzke ◽  
Rudolf F. Guthoff ◽  
Niels Grabow ◽  
Thomas Stahnke
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle Actin ◽  
Human Fibroblasts ◽  
Inhibitory Effect ◽  
Marginal Effect ◽  
Alpha Smooth Muscle Actin ◽  
Primary Fibroblasts ◽  
Implant Coating ◽  
Tgf Β1

Abstract The development of strategies for fibrosis prevention is a perpetual cutting the edge challenge, especially in ophthalmologic fistulating surgery. Stenosis of liquid draining implants but also fibrotic activation of implant-associated tissues are major clinical examples for fibrosis induced implant failure. Implant coating or incorporation of antifibrotic agents offer a promising approach to minimise failure rates. Nintendanib is a drug that has been successfully used for the treatment of ideopathic pulmonary fibrosis since 2015. In this study, we evaluated the suitability of Nintedanib for ophthalmic therapeutic treatment. Therefore, the antifibrotic potential of the active substance was tested with a fibrotic cell culture model on primary fibroblasts of the Tenon (hTF) in vitro. The concentration of 10μM Nintedanib demonstrated a marginal effect on cell viability but coincidently diminished cell proliferation remarkably. Both, immunocytochemical and Western blot analyses revealed a significant inhibitory effect of Nintedanib on the TGF-β1 induced expression of the extracellular matrix (ECM) components fibronectin and collagen. Moreover it supressed the expression and formation of stress fibres of the fibrotic marker protein alpha smooth muscle actin (α-SMA).

scholarly journals RELMα Is Induced in Airway Epithelial Cells by Oncostatin M without Requirement of STAT6 or IL-6 in Mouse Lungs In Vivo

Cells ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 1338 ◽  
Author(s):  
Lilian Ho ◽  
Ashley Yip ◽  
Francis Lao ◽  
Fernando Botelho ◽  
Carl D. Richards
Keyword(s):  
Extracellular Matrix ◽  
Epithelial Cells ◽  
Smooth Muscle Actin ◽  
Airway Epithelial Cells ◽  
Oncostatin M ◽  
Mouse Lung ◽  
Airway Epithelial ◽  
Alpha Smooth Muscle Actin ◽  
Matrix Gene

Resistin-like molecule alpha (RELMα) and YM-1 are secreted proteins implicated in murine models of alternatively activated macrophage (AA/M2) accumulation and Th2-skewed inflammation. Since the gp130 cytokine Oncostatin M (OSM) induces a Th2-like cytokine and AA/M2 skewed inflammation in mouse lung, we here investigated regulation of RELMα and YM-1. Transient pulmonary overexpression of OSM by Adenovirus vector (AdOSM) markedly induced RELMα and YM-1 protein expression in total lung. In situ hybridization showed that RELMα mRNA was highly induced in airway epithelial cells (AEC) and was co-expressed with CD68 mRNA in some but not all CD68+ cells in parenchyma. IL-6 overexpression (a comparator gp130 cytokine) induced RELMα, but at significantly lower levels. IL-6 (assessing IL-6−/− mice) was not required, nor was STAT6 (IL-4/13 canonical signalling) for AdOSM-induction of RELMα in AEC. AEC responded directly to OSM in vitro as assessed by pSTAT3 activation. RELMα-deficient mice showed similar inflammatory cell infiltration and cytokine responses to wt in response to AdOSM, but showed less accumulation of CD206+ AA/M2 macrophages, reduced induction of extracellular matrix gene mRNAs for COL1A1, COL3A1, MMP13, and TIMP1, and reduced parenchymal alpha smooth muscle actin. Thus, RELMα is regulated by OSM in AEC and contributes to extracellular matrix remodelling in mouse lung.

scholarly journals Chronic Cerebral Hypoxia Promotes Arteriogenic Remodeling Events that can be Identified by Reduced Endoglin (CD105) Expression and a Switch in β1 Integrins

2012 ◽  
Vol 32 (9) ◽  
pp. 1820-1830 ◽  
Author(s):  
Amin Boroujerdi ◽  
Jennifer V Welser-Alves ◽  
Ulrich Tigges ◽  
Richard Milner
Keyword(s):  
Endothelial Cells ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Mechanistic Model ◽  
Cerebral Hypoxia ◽  
Alpha Smooth Muscle Actin ◽  
Angiogenic Switch ◽  
Β1 Integrins ◽  
Tgf Β1

Chronic cerebral hypoxia leads to a strong vascular remodeling response, though little is known about which part of the vascular tree is modified, or whether this response includes formation of new arterial vessels. In this study, we examined this process in detail, analyzing how hypoxia (8% O2 for 14 days) alters the size distribution of vessels, number of arteries/arterioles, and expression pattern of endoglin (CD105), a marker of angiogenic endothelial cells in tumors. We found that cerebral hypoxia promoted the biggest increase in the number of medium to large size vessels, and this correlated with increased numbers of alpha smooth muscle actin ( α-SMA)-positive arterial vessels. Surprisingly, hypoxia induced a marked reduction in CD105 expression on brain endothelial cells (BECs) within remodeling arterial vessels, and these BECs also displayed an angiogenic switch in β1 integrins (from α6 to α5), previously described for developmental angiogenesis. In vitro, transforming growth factor (TGF)- β1 also promoted this switch of BEC β1 integrins. Together, these results show that cerebral hypoxia promotes arteriogenesis, and identify reduced CD105 expression as a novel marker of arteriogenesis. Furthermore, our data suggest a mechanistic model whereby BECs in remodeling arterial vessels downregulate CD105 expression, which alters TGF- β1 signaling, to promote a switch in β1 integrins and arteriogenic remodeling.

Abstract 329: Simvastatin Inhibits TGF-β1-induced Proliferation and Activation of Cardiac Fibroblasts Through Inhibition of SMAD Pathway

2014 ◽  
Vol 115 (suppl_1) ◽  
Author(s):  
Farhan Rizvi ◽  
Ramail Siddiqui ◽  
Alesandra DeFranco ◽  
Akhil Jayaprakash ◽  
Mahek Mirza ◽  
...  
Keyword(s):  
Nuclear Translocation ◽  
Cardiac Fibroblasts ◽  
Smooth Muscle Actin ◽  
Fluorescent Microscopy ◽  
Inhibitory Effect ◽  
Myofibroblast Differentiation ◽  
Fibroblast Proliferation ◽  
Alpha Smooth Muscle Actin ◽  
Reductase Inhibitors ◽  
Tgf Β1

Background: HMG-CoA reductase inhibitors (statins) have been shown to reduce the incidence of atrial fibrillation (AF) and its progression but the underlying mechanisms are not fully elucidated. Since atrial fibrosis plays a major role in the development of the substrate for AF progression, we hypothesized that statins antagonize the effect of profibrotic cytokines, reducing their stimulatory effect on fibroblast proliferation, differentiation and activation. Methods: The effect of TGF-β1, a major profibrotic cytokine, on fibroblast proliferation and activation of myofibroblasts was assessed by expression of alpha smooth muscle actin (α-SMA) message (qPCR) and proteins (western and immunofluorocytochemistry) in the presence and absence of simvastatin (1-10μM). The inhibitory effect of simvastatin on SMAD 2/3 phosphorylation (western) and its nuclear translocation by TGF-β1 (5ng) was determined by immunofluorescence antibodies using fluorescent microscopy. Results: TGF-β1 treatment increased fibroblast proliferation (cell count) by 63% compared to control (p<0.001) at 96 hours, which was inhibited by simvastatin by 61% (p<0.001) (Fig a). Simvastatin also reduced TGF-β1-mediated myofibroblast differentiation (α-SMA positive) by 75% (p=0.02); (Fig b and c). TGF-β1 increased SMAD2/3 phosphorylation with increase in nuclear localization was inhibited by simvastatin. Conclusion: Simvastatin inhibits TGFβ-1-mediated cardiac fibroblast proliferation and myofibroblast differentiation by antagonizing SMAD phosphorylation and its translocation into the nucleus.

Systembiologie in der Ophthalmologie – innovative Wirkstoffidentifikation zur spezifischen Vermeidung postoperativer Fibrose

2019 ◽  
Vol 236 (12) ◽  
pp. 1428-1434
Author(s):  
Thomas Stahnke ◽  
Beata Gajda-Derylo ◽  
Oliver Stachs ◽  
Rudolf F. Guthoff ◽  
Anselm Jünemann ◽  
...  
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Transforming Growth Factor Β1 ◽  
Connectivity Map ◽  
Alpha Smooth Muscle Actin ◽  
Next Generation Sequencing Ngs ◽  
Cluster Of Differentiation ◽  
Tgf Β1

Zusammenfassung Hintergrund Der Langzeiterfolg fistulierender Therapiekonzepte zur Behandlung des Glaukoms wird im Wesentlichen durch überschießende Vernarbungsreaktionen (Fibrose) limitiert. Zytostatika wie Mitomycin C können die Fibrose zwar verhindern, sind jedoch häufig mit Nebenwirkungen assoziiert. Spezifisch wirkende Antifibrotika sind derzeit nicht im klinischen Einsatz. Daher beschreibt diese Studie einen systembiologischen Ansatz, mit dem durch eine dedizierte Bioinformatik-Technologieplattform Wirkstoffe identifiziert und als Antifibrotikum repositioniert werden können. Material und Methoden Als Basis für den Wirkstoffidentifikationsprozess wurden differenzielle Genexpressionsdaten humaner Tenon-Fibroblasten (hTF) genutzt, die von unbehandelten hTF und von mit Transforming Growth Factor β1 (TGF-β1) stimulierten hTF („fibrotische Fibroblasten“) mittels Next-Generation Sequencing (NGS) erhoben wurden. Diese Daten wurden mit dem bioinformatischen Werkzeug „FocusHeuristics“ gefiltert. Im Vergleich mit der Connectivity-Map-Datenbank wurden der Fibrose entgegenwirkende Wirkstoffe identifiziert. Die Evaluierung eines potenziell erfolgversprechenden Wirkstoffs als Antifibrotikum wurde an hTF mittels indirekter Immunfluoreszenz in vitro durchgeführt. Ergebnisse Die Analyse der Genexpressionsdaten führte zur Identifikation mehrerer in fibrotische Prozesse involvierter Interaktionsnetzwerke von Genen bzw. Proteinen. Eines dieser Netzwerke beinhaltet das Zytokin Bone morphogenic Protein 6 (BMP6) sowie Interleukin 6 (IL6) und Fibroblast Growth Factor 1 (FGF1). Ein weiteres relevantes Netzwerk konnte rund um das CD34-Gen (CD34: Cluster of Differentiation 34) identifiziert werden. Der Vergleich dieser Daten mit denen der Connectivity Map ermöglichte die Identifikation eines entsprechend invers wirkenden Wirkstoffs. Dessen Evaluierung im fibrotischen Zellkulturmodell in vitro mittels indirekter Immunfluoreszenz führte zu einer deutlichen Expressionsreduktion der fibrotischen Markerproteine Fibronektin und Alpha-smooth Muscle Actin (α-SMA), womit die vorhergesagte antifibrotische Wirkung bestätigt werden konnte. Schlussfolgerung Systembiologische Ansätze können für die Identifikation von antifibrotischen Wirkstoffkandidaten zur Vermeidung postoperativer Fibrose genutzt werden und sollten sich über die Erfassung differenzieller Genexpressionsdaten weiterer okularer Zellen oder Gewebe auch auf andere ophthalmologische Anwendungsfelder transferieren lassen.

Cold Plasma Treatment Promotes Full-thickness Healing of Skin Wounds in Murine Models

2021 ◽  
pp. 153473462110021
Author(s):  
Joon M. Jung ◽  
Hae K. Yoon ◽  
Chang J. Jung ◽  
Soo Y. Jo ◽  
Sang G. Hwang ◽  
...  
Keyword(s):  
Wound Healing ◽  
Plasma Treatment ◽  
Cold Plasma ◽  
Smooth Muscle Actin ◽  
Treated Group ◽  
Control Group ◽  
Alpha Smooth Muscle Actin ◽  
Skin Wound Healing

Cold plasma can be beneficial for promoting skin wound healing and has a high potential of being effectively used in treating various wounds. Our aim was to verify the effect of cold plasma in accelerating wound healing and investigate its underlying mechanism in vitro and in vivo. For the in vivo experiments, 2 full-thickness dermal wounds were created in each mouse (n = 30). While one wound was exposed to 2 daily plasma treatments for 3 min, the other wound served as a control. The wounds were evaluated by imaging and histological analyses at 4, 7, and 11 days post the wound infliction process. Immunohistochemical studies were also performed at the same time points. In vitro proliferation and scratch assay using HaCaT keratinocytes and fibroblasts were performed. The expression levels of wound healing–related genes were analyzed by real-time polymerase chain reaction and western blot analysis. On day 7, the wound healing rates were 53.94% and 63.58% for the control group and the plasma-treated group, respectively. On day 11, these rates were 76.05% and 93.44% for the control and plasma-treated groups, respectively, and the difference between them was significant ( P = .039). Histological analysis demonstrated that plasma treatment promotes the formation of epidermal keratin and granular layers. Immunohistochemical studies also revealed that collagen 1, collagen 3, and alpha-smooth muscle actin appeared more abundantly in the plasma-treated group than in the control group. In vitro, the proliferation of keratinocytes was promoted by plasma exposure. Scratch assay showed that fibroblast exposure to plasma increased their migration. The expression levels of collagen 1, collagen 3, and alpha-smooth muscle actin were elevated upon plasma treatment. In conclusion, cold plasma can accelerate skin wound healing and is well tolerated.

scholarly journals mTOR inhibitors potentially reduce TGF-β2-induced fibrogenic changes in trabecular meshwork cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Nozomi Igarashi ◽  
Megumi Honjo ◽  
Makoto Aihara
Keyword(s):  
Trabecular Meshwork ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Mtor Inhibitors ◽  
In Vivo Model ◽  
Type I ◽  
Fibrotic Response ◽  
Alpha Smooth Muscle Actin

AbstractWe examined the effects of mTOR inhibitors on the fibrotic response induced by transforming growth factor-beta2 (TGF-β2) in cultured human trabecular meshwork (hTM) cells. TGF-β2-induced expression of fibronectin, collagen type I, alpha 1 chain (COL1A1), and alpha-smooth muscle actin (αSMA) in hTM cells was examined in the presence or absence of mTOR inhibitors using quantitative real-time polymerase chain reaction, Western blotting, and immunohistochemistry. The migration rates of hTM cells were examined in the presence of TGF-β2 with or without mTOR inhibitors. An in vitro study showed that the expression of fibronectin, COL1A1, and αSMA was upregulated by TGF-β2 treatment of hTM cells; such upregulation was significantly suppressed by mTOR inhibitors. The inhibitors significantly reduced the migration rate of TGF-β2-stimulated hTM cells. mTOR inhibitors may usefully reduce the fibrotic response of hTM cells and we may have to explore if it is also effective in in vivo model.

scholarly journals A Mouse Model for Studying Stem Cell Effects on Regeneration of Hair Follicle Outer Root Sheaths

2020 ◽  
Vol 15 (1) ◽  
pp. 41-50
Author(s):  
Jingxu Guo ◽  
Shuwei Li ◽  
Hongyang Wang ◽  
Tinghui Wu ◽  
Zhenhui Wu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Mouse Model ◽  
Hair Follicle ◽  
Smooth Muscle Actin ◽  
Hair Follicles ◽  
Alpha Smooth Muscle Actin ◽  
Papillary Structure ◽  
Glass Membrane

AbstractObjectiveStem cells hold promise for treating hair loss. Here an in vitro mouse model was developed using outer root sheaths (ORSs) isolated from hair follicles for studying stem cell-mediated dermal papillary regeneration.MethodsUnder sterile conditions, structurally intact ORSs were isolated from hair follicles of 3-day-old Kunming mice and incubated in growth medium. Samples were collected daily for 5 days. Stem cell distribution, proliferation, differentiation, and migration were monitored during regeneration.ResultsCell proliferation began at the glass membrane periphery then spread gradually toward the membrane center, with the presence of CD34 and CD200 positive stem cells involved in repair initiation. Next, CD34 positive stem cells migrated down the glass membrane, where some participated in ORS formation, while other CD34 cells and CD200 positive cells migrated to hair follicle centers. Within the hair follicle matrix, stem cells divided, grew, differentiated and caused outward expansion of the glass membrane to form a dermal papillary structure containing alpha-smooth muscle actin. Neutrophils attracted to the wound site phagocytosed bacterial and cell debris to protect regenerating tissue from infection.ConclusionIsolated hair follicle ORSs can regenerate new dermal papillary structures in vitro. Stem cells and neutrophils play important roles in the regeneration process.

scholarly journals Endothelial-to-Mesenchymal Transition in Human Adipose Tissue Vasculature Alters the Particulate Secretome and Induces Endothelial Dysfunction

2019 ◽  
Vol 39 (10) ◽  
pp. 2168-2191 ◽  
Author(s):  
Bronson A. Haynes ◽  
Li Fang Yang ◽  
Ryan W. Huyck ◽  
Eric J. Lehrer ◽  
Joshua M. Turner ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Endothelial Dysfunction ◽  
Smooth Muscle Actin ◽  
Phenotypic Change ◽  
Alpha Smooth Muscle Actin ◽  
Mesenchymal Transition ◽  
Molecular Features ◽  
Endothelial To Mesenchymal Transition

Objective: Endothelial cells (EC) in obese adipose tissue (AT) are exposed to a chronic proinflammatory environment that may induce a mesenchymal-like phenotype and altered function. The objective of this study was to establish whether endothelial-to-mesenchymal transition (EndoMT) is present in human AT in obesity and to investigate the effect of such transition on endothelial function and the endothelial particulate secretome represented by extracellular vesicles (EV). Approach and Results: We identified EndoMT in obese human AT depots by immunohistochemical co-localization of CD31 or vWF and α-SMA (alpha-smooth muscle actin). We showed that AT EC exposed in vitro to TGF-β (tumor growth factor-β), TNF-α (tumor necrosis factor-α), and IFN-γ (interferon-γ) undergo EndoMT with progressive loss of endothelial markers. The phenotypic change results in failure to maintain a tight barrier in culture, increased migration, and reduced angiogenesis. EndoMT also reduced mitochondrial oxidative phosphorylation and glycolytic capacity of EC. EVs produced by EC that underwent EndoMT dramatically reduced angiogenic capacity of the recipient naïve ECs without affecting their migration or proliferation. Proteomic analysis of EV produced by EC in the proinflammatory conditions showed presence of several pro-inflammatory and immune proteins along with an enrichment in angiogenic receptors. Conclusions: We demonstrated the presence of EndoMT in human AT in obesity. EndoMT in vitro resulted in production of EV that transferred some of the functional and metabolic features to recipient naïve EC. This result suggests that functional and molecular features of EC that underwent EndoMT in vivo can be disseminated in a paracrine or endocrine fashion and may induce endothelial dysfunction in distant vascular beds.

Expression of alpha-smooth muscle actin, TGF-β1 and TGF-β type II receptor during connective tissue contraction

1997 ◽  
Vol 33 (8) ◽  
pp. 622-627 ◽  
Author(s):  
M. Reza Ghassemifar ◽  
Roy W. Tarnuzzer ◽  
Nasser Chegini ◽  
Erkki Tarpila ◽  
Gregory S. Schultz ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Connective Tissue ◽  
Smooth Muscle Actin ◽  
Type Ii ◽  
Muscle Actin ◽  
Alpha Smooth Muscle Actin ◽  
Tissue Contraction ◽  
Tgf Β1

scholarly journals Activation of transmembrane receptor tyrosine kinase DDR1-STAT3 cascade by extracellular matrix remodeling promotes liver metastatic colonization in uveal melanoma

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Wei Dai ◽  
Shenglan Liu ◽  
Shubo Wang ◽  
Li Zhao ◽  
Xiao Yang ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cancer Cells ◽  
Uveal Melanoma ◽  
Specific Inhibitor ◽  
Type I ◽  
Matrix Remodeling ◽  
Metastatic Colonization ◽  
Tgf Β1

AbstractColonization is believed a rate-limiting step of metastasis cascade. However, its underlying mechanism is not well understood. Uveal melanoma (UM), which is featured with single organ liver metastasis, may provide a simplified model for realizing the complicated colonization process. Because DDR1 was identified to be overexpressed in UM cell lines and specimens, and abundant pathological deposition of extracellular matrix collagen, a type of DDR1 ligand, was noted in the microenvironment of liver in metastatic patients with UM, we postulated the hypothesis that DDR1 and its ligand might ignite the interaction between UM cells and their surrounding niche of liver thereby conferring strengthened survival, proliferation, stemness and eventually promoting metastatic colonization in liver. We tested this hypothesis and found that DDR1 promoted these malignant cellular phenotypes and facilitated metastatic colonization of UM in liver. Mechanistically, UM cells secreted TGF-β1 which induced quiescent hepatic stellate cells (qHSCs) into activated HSCs (aHSCs) which secreted collagen type I. Such a remodeling of extracellular matrix, in turn, activated DDR1, strengthening survival through upregulating STAT3-dependent Mcl-1 expression, enhancing stemness via upregulating STAT3-dependent SOX2, and promoting clonogenicity in cancer cells. Targeting DDR1 by using 7rh, a specific inhibitor, repressed proliferation and survival in vitro and in vivo outgrowth. More importantly, targeting cancer cells by pharmacological inactivation of DDR1 or targeting microenvironmental TGF-β1-collagen I loop exhibited a prominent anti-metastasis effect in mice. In conclusion, targeting DDR1 signaling and TGF-β signaling may be a novel approach to diminish hepatic metastasis in UM.

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