Increased expression of the transforming growth factor β–inducible gene HIC-5 in systemic sclerosis skin and fibroblasts: a novel antifibrotic therapeutic target

Rheumatology ◽  
2020 ◽  
Vol 59 (10) ◽  
pp. 3092-3098 ◽  
Author(s):  
Sonsoles Piera-Velazquez ◽  
Jolanta Fertala ◽  
Gonzalo Huaman-Vargas ◽  
Natalia Louneva ◽  
Sergio A Jiménez
Keyword(s):  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Normal Skin ◽  
Transforming Growth Factor Β ◽  
Dermal Fibroblasts ◽  
Skin Fibroblasts ◽  
Tissue Fibrosis ◽  
Protein Levels ◽  
Antifibrotic Therapy ◽  
Tgf Β1

Abstract Objective SSc is a systemic fibrotic disease affecting skin, numerous internal organs and the microvasculature. The molecular pathogenesis of SSc tissue fibrosis has not been fully elucidated, although TGF-β1 plays a crucial role. The Hic-5 protein encoded by the TGF-β1-inducible HIC-5 gene participates in numerous TGF-β-mediated pathways, however, the role of Hic-5 in SSc fibrosis has not been investigated. The aim of this study was to examine HIC-5 involvement in SSc tissue fibrosis. Methods Affected skin from three patients with diffuse SSc and dermal fibroblasts cultured from affected and non-affected SSc skin were examined for HIC-5 and COL1A1 gene expression. Real-time PCR, IF microscopy, western blotting and small interfering RNA–mediated HIC-5 were performed. Results HIC-5 and COL1A1 transcripts and Hic-5, type 1 collagen (COL1) and α-smooth muscle actin (α-SMA) protein levels were increased in clinically affected SSc skin compared with normal skin and in cultured dermal fibroblasts from affected SSc skin compared with non-affected skin fibroblasts from the same patients. HIC-5 knockdown caused a marked reduction of COL1 production in SSc dermal fibroblasts. Conclusion HIC-5 expression is increased in affected SSc skin compared with skin from normal individuals. Affected SSc skin fibroblasts display increased HIC-5 and COL1A1 expression compared with non-affected skin fibroblasts from the same patients. Hic-5 protein was significantly increased in cultured SSc dermal fibroblasts. HIC-5 mRNA knockdown in SSc fibroblasts caused >50% reduction of COL1 production. Although these are preliminary results owing to the small number of skin samples studied, they indicate that Hic-5 plays a role in the profibrotic activation of SSc dermal fibroblasts and may represent a novel molecular target for antifibrotic therapy in SSc.

TAT-mediated PRDX6 protein transduction protects against eye lens epithelial cell death and delays lens opacity

2008 ◽  
Vol 294 (3) ◽  
pp. C842-C855 ◽  
Author(s):  
Eri Kubo ◽  
Nigar Fatma ◽  
Yoshio Akagi ◽  
David R. Beier ◽  
Sanjay P. Singh ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Lens Epithelial Cell ◽  
Biologically Active ◽  
Lens Epithelial Cells ◽  
Protein Levels ◽  
Endogenous Antioxidant ◽  
Tgf Β1

A diminished level of endogenous antioxidant in cells/tissues is associated with reduced resistance to oxidative stress. Peroxiredoxin 6 (PRDX6), a protective molecule, regulates gene expression/function by controlling reactive oxygen species (ROS) levels. Using PRDX6 protein linked to TAT, the transduction domain from human immunodeficiency virus type 1 TAT protein, we demonstrated that PRDX6 was transduced into lens epithelial cells derived from rat or mouse lenses. The protein was biologically active, negatively regulating apoptosis and delaying progression of cataractogenesis by attenuating deleterious signaling. Lens epithelial cells from cataractous lenses bore elevated levels of ROS and were susceptible to oxidative stress. These cells harbored increased levels of active transforming growth factor (TGF)-β1 and of α-smooth muscle actin and βig-h3, markers for cataractogenesis. Importantly, cataractous lenses showed a 10-fold reduction in PRDX6 expression, whereas TGF-β1 mRNA and protein levels were elevated. The changes were reversed, and cataractogenesis was delayed when PRDX6 was supplied. Results suggest that delivery of PRDX6 can postpone cataractogenesis, and this should be an effective approach to delaying cataracts and other degenerative diseases that are associated with increased ROS.

scholarly journals TGF-β1/SMOC2/AKT and ERK axis regulates proliferation, migration, and fibroblast to myofibroblast transformation in lung fibroblast, contributing with the asthma progression

Hereditas ◽  
2021 ◽  
Vol 158 (1) ◽  
Author(s):  
Yuebin Wang ◽  
Huike Yang ◽  
Xian Su ◽  
Anqiang Cao ◽  
Feng Chen ◽  
...  
Keyword(s):  
Calcium Binding ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Transforming Growth Factor Β ◽  
Chronic Respiratory Disease ◽  
Lung Fibroblasts ◽  
Promoting Effect ◽  
And Migration ◽  
Tgf Β1 ◽  
Proliferation And Migration

Abstract Background Asthma is a common chronic respiratory disease that influences 300 million people all over the world. However, the pathogenesis of asthma has not been fully elucidated. It has been reported that transforming growth factor-β (TGF-β) can activate myofibroblasts. Moreover, the fibroblast to myofibroblast transformation (FMT) can be triggered by TGF-β, which is a major mediator of subepithelial fibrosis. Secreted modular calcium-binding protein 2 (SMOC2) is a member of cysteine (SPARC) family and is involved in the progression of multiple diseases. However, its role in asthma remains poorly understood. RT-qPCR evaluated the expression of SMOC2. Bromodeoxyuridine assay and wound-healing assay detected the proliferation and migration of lung fibroblasts, respectively. IF staining was performed to assess the expression of α-smooth muscle actin (α-SMA). Western blot analysis detected the levels of proteins. Flow cytometry was utilized for determination of the number of myofibroblasts. Results We found the expression of SMOC2 was upregulated by the treatment of TGF-β1 in lung fibroblasts. In addition, SMOC2 promoted the proliferation and migration of lung fibroblasts. More importantly, SMOC2 accelerated FMT of lung fibroblasts. Furthermore, SMOC2 was verified to control the activation of AKT and ERK. Rescue assays showed that the inhibition of AKT and ERK pathway reversed the promoting effect of SMOC2 overexpression on proliferation, migration and FMT in lung fibroblasts. Conclusions This work demonstrated that SMOC2 modulated TGF-β1-induced proliferation, migration and FMT in lung fibroblasts and may promote asthma, which potentially provided a novel therapeutic target for the management of asthma.

scholarly journals Protective effects of Pai-Du-Yang-Shen formula on chronic renal failure in rats

2019 ◽  
Vol 17 ◽  
pp. 205873921987141
Author(s):  
Ronghua Pan ◽  
Guohua Rui ◽  
Xiaohui Bai ◽  
Zhiyin Teng ◽  
Gang Chen ◽  
...  
Keyword(s):  
Renal Failure ◽  
Chronic Renal Failure ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Transforming Growth Factor Β ◽  
Therapeutic Effects ◽  
Protective Effects ◽  
Protein Levels ◽  
Yang Shen ◽  
Necrosis Factor

This study aimed to elucidate the therapeutic effects of a traditional Chinese medicine, Pai-Du-Yang-Shen (PDYS) Formula, in 5/6 nephrectomy-induced chronic renal failure (CRF). The CRF model was established via 5/6 nephrectomy (Nx) in rats. Then, PDYS (0.9 and 1.8 g/kg/day), or an equal amount of normal saline, was administrated to the rats in respective groups. Our results showed that PDYS treatment improved degeneration, fibrosis, and histopathological abnormalities of renal tissues in 5/6 Nx rats. Furthermore, the increase in serum creatinine (Scr), blood urea nitrogen (BUN), and total urinary protein (TUP) in 5/6 Nx rats was reduced by PDYS treatment. In addition, the levels of tumor necrosis factor (TNF), fibronectin (FN), and laminin (LN) that were induced in 5/6 Nx rats were also decreased by PDYS treatment. Finally, relative mRNA expression of α-smooth muscle actin (α-SMA), FN, transforming growth factor-β (TGF-β), and TNF as well as protein levels of TGF-β, NF-κB p65, and phosphorylated NF-κB p65 (p-p65) in renal tissues correlated with the concentration of TNF. In conclusion, PDYS shows therapeutic effects against CRF as determined by a reduction in markers for inflammation and fibrosis.

Tissue-type plasminogen activator deficiency attenuates peritoneal fibrosis in mice

2009 ◽  
Vol 297 (6) ◽  
pp. F1510-F1517 ◽  
Author(s):  
Kei Kurata ◽  
Shoichi Maruyama ◽  
Sawako Kato ◽  
Waichi Sato ◽  
Jun-ichiro Yamamoto ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Basement Membranes ◽  
Tissue Type ◽  
Vascular Density ◽  
Peritoneal Fibrosis ◽  
Plasmin Activity ◽  
Protein Levels ◽  
Tgf Β1

Peritoneal fibrosis (PF) is an important complication of peritoneal dialysis therapy. The present study was performed to examine the mechanisms of PF in view of the plasminogen activator (PA)/plasmin/matrix metalloproteinase (MMP) cascade. PF was induced in tissue-type PA (tPA) deficient mice and wild-type mice by intraperitoneal injection of chlorhexidine gluconate. Mice were killed on day 21, and tissue samples were taken. Histopathological studies were performed. Plasmin activity, gelatinases activity, and the levels of tPA, transforming growth factor-β1 (TGF-β1), and MMP-2 mRNA were determined. Protein levels of MMP-3, tissue inhibitor of metalloproteinases (TIMP)-1, -2, and -3, phospho-Smad3, membrane-type 1 (MT1)-MMP, and MT3-MMP were also studied. On day 21, tPA +/+ mice showed severe PF, whereas tPA −/− mice showed milder change. Submesothelial basement membranes were dissolved in tPA +/+ mice while they were relatively preserved in tPA −/− mice. The levels of macrophage infiltration, staining for α-smooth muscle actin (α-SMA) and collagen type III, and vascular density were all significantly lower in tPA −/− mice than in tPA +/+ mice. The levels of plasmin activity, pro- and active MMP-2, mRNA expression of tPA and TGF-β1, and phospho-Smad3 protein were also lower in tPA −/− mice. No difference was observed between the two groups concerning the protein levels of MMP-3, TIMP-1, TIMP-2, TIMP-3, MT1-MMP, or MT3-MMP. These results indicate that the presence of tPA enhances inflammation, angiogenesis, and fibrogenesis in the peritoneum of the PF model mice. Activation of the PA/plasmin/MMP cascade may play a pivotal role in the pathogenesis of PF.

scholarly journals Bone Morphogenetic Protein Antagonist Gremlin-1 Increases Myofibroblast Transition in Dermal Fibroblasts: Implications for Systemic Sclerosis

2021 ◽  
Vol 9 ◽  
Author(s):  
Laura Duffy ◽  
John Henderson ◽  
Max Brown ◽  
Stefan Pryzborski ◽  
Nicola Fullard ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Smooth Muscle Actin ◽  
Mapk Signaling ◽  
Dermal Fibroblasts ◽  
Expression Vectors ◽  
Developmentally Regulated ◽  
Protein Levels ◽  
Morphogenetic Protein ◽  
Tgf Β1 ◽  
Gremlin 1

ObjectiveSystemic Sclerosis is an autoimmune connective tissue disease which results in fibrosis of the skin and lungs. The disease is characterized by activation of myofibroblasts but what governs this is unknown. Gremlin-1 is a BMP antagonist that is developmentally regulated and we sought to investigate its role in Systemic Sclerosis.MethodsDermal fibroblasts were transfected with Grem1pcDNA3.1 expression vectors or empty vectors. Various markers of myofibroblasts were measured at the mRNA and protein levels. Scratch wound assays were also performed. Media Transfer experiments were performed to evaluate cytokine like effects. Various inhibitors of TGF-β signaling and MAPK signaling were used post-transfection. siRNA to Gremlin-1 in SSc dermal fibroblasts were performed to evaluate the role of Gremlin-1. Different cytokines were incubated with fibroblasts and Gremlin-1 measured. Bleomycin was used as model of fibrosis and immunohistochemistry performed.ResultsOverexpression of Gremlin-1 was achieved in primary dermal fibroblasts and lead to activation of quiescent cells to myofibroblasts indicated by collagen and α-Smooth muscle actin. Overexpression also led to functional effects. This was associated with increased TGF-β1 levels and SBE luciferase activity but not increased Thrombospondin-1 expression. Inhibition of Gremlin-1 overexpression cells with antibodies to TGF-β1 but not isotype controls led to reduced collagen and various TGF-β pathway chemical inhibitors also led to reduced collagen levels. In SSc cells siRNA mediated reduction of Gremlin-1 reduced collagen expression and CTGF gene and protein levels in these cells. IL-13 did not lead to elevated Gremlin-1 expression nor did IL-11. Gremlin-1 was elevated in an animal model of fibrosis compared to NaCl-treated mice.ConclusionGremlin-1 is a key regulator of myofibroblast transition leading to enhanced ECM deposition. Strategies that block Gremlin-1 maybe a possible therapeutic target in fibrotic diseases such as SSc.

scholarly journals Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway

2021 ◽  
Vol 12 ◽  
Author(s):  
Bo Zhang ◽  
Lihua Gao ◽  
Chunsheng Shao ◽  
Mingsi Deng ◽  
Liangjian Chen
Keyword(s):  
Growth Factor ◽  
Signaling Pathway ◽  
Transforming Growth Factor ◽  
Clinical Investigation ◽  
Smooth Muscle Actin ◽  
Oral Submucous Fibrosis ◽  
Transforming Growth Factor Β ◽  
Type I ◽  
Camp Analog ◽  
Tgf Β1

Chewing areca nut (betel quid) is strongly associated with oral submucous fibrosis (OSF), a pre-cancerous lesion. Among the areca alkaloids, arecoline is the main agent responsible for fibroblast proliferation; however, the specific molecular mechanism of arecoline affecting the OSF remains unclear. The present study revealed that arecoline treatment significantly enhanced Transforming growth factor-β (TGF-β)-induced buccal mucosal fibroblast (BMF) activation and fibrotic changes. Arecoline interacts with phosphodiesterase 4A (PDE4A) to exert its effects through modulating PDE4A activity but not PDE4A expression. PDE4A silence reversed the effects of arecoline on TGF-β-induced BMFs activation and fibrotic changes. Moreover, the exchange protein directly activated by cAMP 1 (Epac1)-selective Cyclic adenosine 3′,5′-monophosphate (cAMP) analog (8-Me-cAMP) but not the protein kinase A (PKA)-selective cAMP analog (N6-cAMP) remarkably suppressed α-smooth muscle actin(α-SMA) and Collagen Type I Alpha 1 Chain (Col1A1) protein levels in response to TGF-β1 and arecoline co-treatment, indicating that cAMP-Epac1 but not cAMP-PKA signaling is involved in arecoline functions on TGF-β1-induced BMFs activation. In conclusion, arecoline promotes TGF-β1-induced BMFs activation through enhancing PDE4A activity and the cAMP-Epac1 signaling pathway during OSF. This novel mechanism might provide more powerful strategies for OSF treatment, requiring further in vivo and clinical investigation.

scholarly journals Relationship between Axial Length and Levels of TGF-β in the Aqueous Humor and Plasma of Myopic Patients

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Yuwen Hsiao ◽  
Yiting Cao ◽  
Yu Yue ◽  
Jibo Zhou
Keyword(s):  
Aqueous Humor ◽  
Axial Length ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Cross Sectional Study ◽  
Cross Sectional ◽  
Protein Levels ◽  
Group A ◽  
Group B ◽  
Tgf Β1

Purpose. To investigate the levels of transforming growth factor-β (TGF-β) in human aqueous humor (AH) and plasma (PL) of patients with myopia, and verify whether there is an association between these levels and their association with axial length (AL). Methods. Thirty-eight myopic patients who received intraocular collamer lens (ICL) implantation were enrolled in this cross-sectional study. Patients were divided into three groups based on AL with cut-off points of 26 and 28 mm. AH and PL samples were obtained during ICL implantation surgery. The levels of TGF-β1, TGF-β2, and TGF-β3 in the AH and PL samples were measured using Luminex xMAP Technology kits (Milliplex xMAP kits). The protein levels of TGF-βs in both AH and PL samples and their relationships with AL were analyzed. Results. In all, 38 patients (59 eyes) were enrolled and divided into the three groups: group A contained 7 people (10 eyes), group B contained 22 people (37 eyes), and group C contained 9 people (12 eyes). In the AH group, we detected TGF-β1 ( P 50 : 19.97 pg/mL), TGF-β2 (2446.00 pg/mL), and TGF-β3 (26.33 pg/mL); in PL, these concentrations were 8984.00, 523.44, and 210.47 pg/mL, respectively. The levels of TGF-β1 and TGF-β3 in AH were positively associated with AL. None of the three isoforms in PL were related to those in AH or to AL. Conclusions. The levels of TGF-β1 and TGF-β3 in AH were more strongly associated with the severity of myopia than the types of TGF-β in PL.

scholarly journals miR-218 regulates focal adhesion kinase–dependent TGFβ signaling in fibroblasts

2014 ◽  
Vol 25 (7) ◽  
pp. 1151-1158 ◽  
Author(s):  
Fen Guo ◽  
David E. Carter ◽  
Andrew Leask
Keyword(s):  
Smooth Muscle ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Transforming Growth Factor Β ◽  
Dermal Fibroblasts ◽  
Muscle Actin ◽  
Gingival Fibroblasts ◽  
Remodeling Of Extracellular Matrix

Scarring, which occurs in essentially all adult tissue, is characterized by the excessive production and remodeling of extracellular matrix by α-smooth muscle actin (SMA)–expressing myofibroblasts located within connective tissue. Excessive scarring can cause organ failure and death. Oral gingivae do not scar. Compared to dermal fibroblasts, gingival fibroblasts are less responsive to transforming growth factor β (TGFβ) due to the reduced expression, due to the reduced expression and activity of focal adhesion kinase (FAK) by this cell type. Here we show that, compared with dermal fibroblasts, gingival fibroblasts show reduced expression of miR-218. Introduction of pre–miR-218 into gingival fibroblasts elevates FAK expression and, via a FAK/src-dependent mechanism, results in the ability of TGFβ to induce α-SMA. The deubiquitinase cezanne is a direct target of miR-218 and has increased expression in gingival fibroblasts compared with dermal fibroblasts. Knockdown of cezanne in gingival fibroblasts increases FAK expression and causes TGFβ to induce α-smooth muscle actin (α-SMA). These results suggest that miR-218 regulates the ability of TGFβ to induce myofibroblast differentiation in fibroblasts via cezanne/FAK.

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