Acute Hyperthermia Inhibits TGF-β1-induced Cardiac Fibroblast Activation via Suppression of Akt Signaling

BACKGROUND: Following myocardial infarction (MI), activated cardiac myofibroblasts facilitate extracellular matrix (ECM) remodeling to prevent mechanical complications. However, prolonged myofibroblast activity leads to dysregulation of the ECM, maladaptive remodeling, fibrosis and heart failure (HF). Chronic inflammation is believed to drive persistent myofibroblast activity, however, the mechanisms are unclear. In this study, we explored the effects of peripheral blood monocytes on human cardiac fibroblast activation in a 3D ECM microenvironment. METHODS/RESULTS: Human cardiac fibroblasts isolated from surgical human heart biopsies were seeded into 3D collagen matrices. Peripheral blood monocytes isolated from healthy human donors were co-cultured with fibroblasts. Monocytes increased fibroblast activation measured by collagen ECM contraction (17.9±11.1% increase; p<0.01) and resulted in local ECM remodeling observed by confocal microscopy. Under co-culture conditions that prevent cell-cell contact but allow interaction via paracrine factors, monocytes had minimal effects on fibroblast activation (6.4±7.0 vs.17.9±11.1% increase, respectively; p<0.01). Multiplex analysis of the co-culture media revealed an increase in the paracrine factors Transforming Growth Factor-beta 1 (TGF-β1) and Matrix Metalloproteinase 9 when monocytes and fibroblasts were cultured under cell-cell contact conditions (162.2±11.7pg/mL and 17.5±0.5ng/mL, respectively, vs. 21.8±5.7pg/mL and 4.9 ±0.4ng/mL; p<0.001). TGF-β1 blockade abolished monocyte induced cardiac fibroblast activation, as did β1-integrin. These data suggest direct cell-cell interaction between monocytes and cardiac fibroblasts through β1-integrin results in TGF-β1 release facilitating fibroblast activation and matrix remodeling. CONCLUSION: For the first time, we demonstrate that peripheral blood monocytes stimulate human cardiac fibroblast activation through a mechanism involving TGF-β1 release as a consequence of direct cell-cell interaction through β1-integrin. These data implicate inflammation as a driver of cardiac fibrosis post-MI, highlighting potential novel therapeutic targets for the treatment of ischemic HF.

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TGF-β1-containing exosomes from cardiac microvascular endothelial cells mediate cardiac fibroblast activation under high glucose conditions

Biochemistry and Cell Biology ◽

10.1139/bcb-2020-0624 ◽

2021 ◽

pp. 1-7

Author(s):

Yan Zhang ◽

Zhengru Zhu ◽

Tingting Wang ◽

Yuan Dong ◽

Yanhong Fan ◽

...

Keyword(s):

Endothelial Cells ◽

High Glucose ◽

Interstitial Fibrosis ◽

Cardiac Fibroblast ◽

Microvascular Endothelial Cells ◽

Ecm Remodeling ◽

Fibroblast Activation ◽

Microvascular Endothelial ◽

Cardiac Microvascular Endothelial Cells ◽

Tgf Β1

Cardiac fibroblast (CF)-mediated extracellular matrix (ECM) remodeling is the key pathological basis for the occurrence and development of diabetic cardiomyopathy (DCM); its specific regulatory mechanisms have been widely studied but remain unclear. Exosomes are a type of stable signal transmission medium, and exosome-mediated cell-cell interactions play an important role in DCM. Endothelial cells form an important barrier between circulation and cardiomyocytes, in addition to being an important endocrine organ of the heart and an initial target for hyperglycemia, a key aspect in the development of DCM. We previously showed that exosomes derived from cardiac microvascular endothelial cells (CMECs) under high glucose conditions can be taken up by cardiomyocytes and regulate autophagy, apoptosis, and glucose metabolism. Consequently, in the present study, we focused on how exosomes mediate the interaction between CMECs and CFs. Surprisingly, exosomes derived from CMECs under high glucose were rich in TGF-β1 mRNA, which significantly promoted the activation of CFs. Additionally, exosomes derived from CMECs under high glucose conditions aggravated perivascular and interstitial fibrosis in mice treated with streptozotocin. Herein, we demonstrated for the first time the capacity of exosomes, released by CMECs under high glucose, to mediate fibroblast activation through TGF-β1 mRNA, which may be potentially beneficial in the development of exosome-targeted therapies to control DCM.

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LncRNA LPAL2/miR-1287-5p/EGFR axis modulates TED derived orbital fibroblast activation through cell adhesion factors

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/clinem/dgab256 ◽

2021 ◽

Author(s):

Nuo Wang ◽

Shi-ying Hou ◽

Xin Qi ◽

Mi Deng ◽

Jia-min Cao ◽

...

Keyword(s):

Cell Adhesion ◽

Akt Signaling ◽

Thyroid Eye Disease ◽

Differentially Expressed ◽

Eye Disease ◽

Fibroblast Activation ◽

Orbital Fibroblast ◽

Orbital Fibroblasts ◽

Adhesion Factor ◽

Tgf Β1

Abstract Background and aims The activation of orbital fibroblasts, the prime targets in thyroid eye disease, is central to its underlying pathogenesis. We aimed to investigate the mechanism of thyroid eye disease orbital fibroblast activation from the perspective of non-coding RNA regulation. Methods Immunofluorescence (IF) staining was applied to evaluate the fibrotic changes in target cells. Cell proliferation were evaluated by EDU and colony formation assays. Collagen I concentration was determined by ELISA assay. Human microarray analysis was performed on three thyroid eye disease and 3 healthy control orbital tissue samples. Results Bioinformatics analysis showed that cell adhesion signaling factors were differentially expressed in thyroid eye disease tissues, including I-CAM-1, I-CAM-4, V-CAM, and CD44, which were all upregulated in diseased orbital tissues. LncRNA LPAL2 level was also upregulated in orbital tissues and positively correlated with I-CAM-1 and I-CAM-4 expression. Stimulation of the thyroid eye disease orbital fibroblasts by TGF-β1 significantly increased the expression of I-CAM-1, I-CAM-4, and LPAL2. Knockdown of LPAL2 in orbital fibroblasts inhibited TGF-β1-induced increases in cell adhesion factor levels and orbital fibroblast activation. Microarray profiling was performed on thyroid eye disease and normal orbital tissues to identify differentially expressed miRNAs and miR-1287-5p was remarkably reduced within diseased orbital samples. miR-1287-5p was directly bound to EGFR 3’UTR and LPAL2 and LPAL2 modulated EGFR/AKT signaling through targeting miR-1287-5p. Conclusions The LPAL2/miR-1287-5p axis modulated TGF-β1-induced increases in cell adhesion factor levels and thyroid eye disease orbital fibroblast activation through EGFR/AKT signaling.

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Effects of emodin, a plant‐derived anthraquinone, on TGF‐β1‐induced cardiac fibroblast activation and function

Journal of Cellular Physiology ◽

10.1002/jcp.30416 ◽

2021 ◽

Author(s):

Wayne Carver ◽

Ethan Fix ◽

Charity Fix ◽

Daping Fan ◽

Mrinmay Chakrabarti ◽

...

Keyword(s):

Cardiac Fibroblast ◽

Fibroblast Activation ◽

And Function ◽

Tgf Β1

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Effects of the isothiocyanate sulforaphane on TGF‐β1‐induced rat cardiac fibroblast activation and extracellular matrix interactions

Journal of Cellular Physiology ◽

10.1002/jcp.28075 ◽

2019 ◽

Vol 234 (8) ◽

pp. 13931-13941 ◽

Cited By ~ 8

Author(s):

Charity Fix ◽

Amanda Carver‐Molina ◽

Mrinmay Chakrabarti ◽

Mohamad Azhar ◽

Wayne Carver

Keyword(s):

Extracellular Matrix ◽

Cardiac Fibroblast ◽

Fibroblast Activation ◽

Matrix Interactions ◽

Extracellular Matrix Interactions ◽

Tgf Β1

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Interstitial fluid flow and cyclic strain differentially regulate cardiac fibroblast activation via AT1R and TGF-β1

Experimental Cell Research ◽

10.1016/j.yexcr.2011.10.008 ◽

2012 ◽

Vol 318 (1) ◽

pp. 75-84 ◽

Cited By ~ 27

Author(s):

P.A. Galie ◽

M.W. Russell ◽

M.V. Westfall ◽

J.P. Stegemann

Keyword(s):

Fluid Flow ◽

Interstitial Fluid ◽

Cyclic Strain ◽

Cardiac Fibroblast ◽

Interstitial Fluid Flow ◽

Fibroblast Activation ◽

Tgf Β1

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Pellino1-mediated TGF-β1 synthesis contributes to mechanical stress induced cardiac fibroblast activation

Journal of Molecular and Cellular Cardiology ◽

10.1016/j.yjmcc.2014.11.006 ◽

2015 ◽

Vol 79 ◽

pp. 145-156 ◽

Cited By ~ 31

Author(s):

Juan Song ◽

Yun Zhu ◽

Jiantao Li ◽

Jiahao Liu ◽

Yun Gao ◽

...

Keyword(s):

Mechanical Stress ◽

Cardiac Fibroblast ◽

Fibroblast Activation ◽

Tgf Β1

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Apigenin Alleviates Renal Fibroblast Activation through AMPK and ERK Signaling Pathways In Vitro

Current Pharmaceutical Biotechnology ◽

10.2174/1389201021666200320140908 ◽

2020 ◽

Vol 21 (11) ◽

pp. 1107-1118

Author(s):

Ningning Li ◽

Zhan Wang ◽

Tao Sun ◽

Yanfei Lei ◽

Xianghua Liu ◽

...

Keyword(s):

Renal Fibrosis ◽

Transforming Growth Factor ◽

Therapeutic Potential ◽

Protective Role ◽

Fibroblast Proliferation ◽

Fibroblast Activation ◽

And Function ◽

Activated Fibroblasts ◽

Tgf Β1

Objective: Renal fibrosis is a common pathway leading to the progression of chronic kidney disease. Activated fibroblasts contribute remarkably to the development of renal fibrosis. Although apigenin has been demonstrated to play a protective role from fibrotic diseases, its pharmacological effect on renal fibroblast activation remains largely unknown. Materials and Methods: Here, we examined the functional role of apigenin in the activation of renal fibroblasts response to transforming growth factor (TGF)-β1 and its potential mechanisms. Cultured renal fibroblasts (NRK-49F) were exposed to apigenin (1, 5, 10 and 20 μM), followed by the stimulation of TGF-β1 (2 ng/mL) for 24 h. The markers of fibroblast activation were determined. In order to confirm the anti-fibrosis effect of apigenin, the expression of fibrosis-associated genes in renal fibroblasts was assessed. As a consequence, apigenin alleviated fibroblast proliferation and fibroblastmyofibroblast differentiation induced by TGF-β1. Result: Notably, apigenin significantly inhibited the fibrosis-associated genes expression in renal fibroblasts. Moreover, apigenin treatment significantly increased the phosphorylation of AMP-activated protein kinase (AMPK). Apigenin treatment also obviously reduced TGF-β1 induced phosphorylation of ERK1/2 but not Smad2/3, p38 and JNK MAPK in renal fibroblasts. Conclusion: In a summary, these results indicate that apigenin inhibits renal fibroblast proliferation, differentiation and function by AMPK activation and reduced ERK1/2 phosphorylation, suggesting it could be an attractive therapeutic potential for the treatment of renal fibrosis.

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