Influence of cytokines and growth factors in ANG II-mediated collagen upregulation by fibroblasts in rats: role of myocytes

Abnormal stiffness and altered cardiac function arising from abnormal collagen deposition occur in hypertrophy and heart failure. ANG II has been shown to play a role in this process. To evaluate the mechanism, we developed an in vitro model by subjecting fibroblasts to ANG II treatment in the presence or absence of myocytes in coculture ( 25 ). Employing this model, we demonstrated that ANG II-induced collagen gene transcription in cardiac fibroblasts was potentiated by myocyte-derived factors. In attempting to identify mechanisms of collagen upregulation and to define the role of myocytes, we found that interleukin (IL)-6, tumor necrosis factor (TNF)-α, and the transforming growth factor (TGF)-β superfamily were also involved in collagen upregulation. Collagen transcripts were increased after fibroblasts were treated with IL-6 (20–50 ng/ml) and TNF-α (0.1–0.5 ng/ml). In this study, we show that cardiomyocytes induce secretion of active TGF-β in the presence of ANG II and that a paracrine action of TGF-β subsequently induces different cytokines (IL-6) in fibroblasts, thereby promoting collagen synthesis. The cross-talk between myocytes and fibroblasts and involvement of these cytokines in the upregulation of collagen transcript levels are novel findings that may explain their possible roles in the upregulation of collagen.

Download Full-text

GSK-3 mediates differentiation and activation of proinflammatory dendritic cells

Blood ◽

10.1182/blood-2006-06-028951 ◽

2006 ◽

Vol 109 (4) ◽

pp. 1584-1592 ◽

Cited By ~ 82

Author(s):

Elena Rodionova ◽

Michael Conzelmann ◽

Eugene Maraskovsky ◽

Michael Hess ◽

Michael Kirsch ◽

...

Keyword(s):

Dendritic Cells ◽

In Vitro Model ◽

Glycogen Synthase ◽

Human Monocyte ◽

Glycogen Synthase Kinase 3 ◽

Inhibitory Effects ◽

Tnf Α ◽

Vitro Model

Abstract The key components of the intracellular molecular network required for the expression of a specific function of dendritic cells (DCs) are as yet undefined. Using an in vitro model of human monocyte-derived DC differentiation, this study investigates the role of glycogen synthase kinase 3 (GSK-3), a multifunctional enzyme critical for cellular differentiation, apoptosis, self-renewal, and motility, in this context. We demonstrate that GSK-3 (1) inhibits macrophage development during differentiation of DCs, (2) is constitutively active in immature DCs and suppresses spontaneous maturation, and (3) acquires a proinflammatory functional status mediating high levels of IL-12, IL-6, and TNF-α secretion, and partially inhibits IL-10 in the context of DC activation. In particular, GSK-3 enhances IL-12p35 mRNA expression and thus the production of the proinflammatory cytokine IL-12p70 by integrating the activities of other kinases priming GSK-3 targets and the inhibitory effects of Akt-1. GSK-3 may therefore act as a key integrator of activating and inhibitory pathways involved in proinflammatory DC differentiation and activation.

Download Full-text

Modulation of Adhesion Process, E-Selectin and VEGF Production by Anthocyanins and Their Metabolites in an In Vitro Model of Atherosclerosis

Nutrients ◽

10.3390/nu12030655 ◽

2020 ◽

Vol 12 (3) ◽

pp. 655 ◽

Cited By ~ 1

Author(s):

Mirko Marino ◽

Cristian Del Bo’ ◽

Massimiliano Tucci ◽

Dorothy Klimis-Zacas ◽

Patrizia Riso ◽

...

Keyword(s):

Endothelial Cells ◽

In Vitro Model ◽

Vascular Endothelial ◽

Tnf Α ◽

Vitro Model ◽

Factor Α ◽

Endothelial Growth Factor ◽

Adhesion Process ◽

Necrosis Factor

The present study aims to evaluate the ability of peonidin and petunidin-3-glucoside (Peo-3-glc and Pet-3-glc) and their metabolites (vanillic acid; VA and methyl-gallic acid; MetGA), to prevent monocyte (THP-1) adhesion to endothelial cells (HUVECs), and to reduce the production of vascular cell adhesion molecule (VCAM)-1, E-selectin and vascular endothelial growth factor (VEGF) in a stimulated pro-inflammatory environment, a pivotal step of atherogenesis. Tumor necrosis factor-α (TNF-α; 100 ng mL−1) was used to stimulate the adhesion of labelled monocytes (THP-1) to endothelial cells (HUVECs). Successively, different concentrations of Peo-3-glc and Pet-3-glc (0.02 µM, 0.2 µM, 2 µM and 20 µM), VA and MetGA (0.05 µM, 0.5 µM, 5 µM and 50 µM) were tested. After 24 h, VCAM-1, E-selectin and VEGF were quantified by ELISA, while the adhesion process was measured spectrophotometrically. Peo-3-glc and Pet-3-glc (from 0.02 µM to 20 µM) significantly (p < 0.0001) decreased THP-1 adhesion to HUVECs at all concentrations (−37%, −24%, −30% and −47% for Peo-3-glc; −37%, −33%, −33% and −45% for Pet-3-glc). VA, but not MetGA, reduced the adhesion process at 50 µM (−21%; p < 0.001). At the same concentrations, a significant (p < 0.0001) reduction of E-selectin, but not VCAM-1, was documented. In addition, anthocyanins and their metabolites significantly decreased (p < 0.001) VEGF production. The present findings suggest that while Peo-3-glc and Pet-3-glc (but not their metabolites) reduced monocyte adhesion to endothelial cells through suppression of E-selectin production, VEGF production was reduced by both anthocyanins and their metabolites, suggesting a role in the regulation of angiogenesis.

Download Full-text

An in vitro model of human bronchial epithelial cells for the investigation of the role of the airway epithelium in asthma exacerbations

Pneumologie ◽

10.1055/s-0037-1619393 ◽

2018 ◽

Vol 72 (S 01) ◽

pp. S101-S102

Author(s):

JC Ehlers ◽

S Bartel ◽

C Vock ◽

H Fehrenbach

Keyword(s):

Epithelial Cells ◽

Airway Epithelium ◽

In Vitro Model ◽

Bronchial Epithelial Cells ◽

Human Bronchial Epithelial Cells ◽

Bronchial Epithelial ◽

Asthma Exacerbations ◽

Vitro Model

Download Full-text

Identification of a microRNA signature in renal fibrosis: role of miR-21

AJP Renal Physiology ◽

10.1152/ajprenal.00273.2011 ◽

2011 ◽

Vol 301 (4) ◽

pp. F793-F801 ◽

Cited By ~ 169

Author(s):

Abolfazl Zarjou ◽

Shanzhong Yang ◽

Edward Abraham ◽

Anupam Agarwal ◽

Gang Liu

Keyword(s):

Epithelial Cells ◽

Renal Fibrosis ◽

Transforming Growth Factor ◽

Response To Treatment ◽

Tubular Epithelial Cells ◽

Altered Expression ◽

Tnf Α ◽

Tgf Β1

Renal fibrosis is a final stage of many forms of kidney disease and leads to impairment of kidney function. The molecular pathogenesis of renal fibrosis is currently not well-understood. microRNAs (miRNAs) are important players in initiation and progression of many pathologic processes including diabetes, cancer, and cardiovascular disease. However, the role of miRNAs in kidney injury and repair is not well-characterized. In the present study, we found a unique miRNA signature associated with unilateral ureteral obstruction (UUO)-induced renal fibrosis. We found altered expression in UUO kidneys of miRNAs that have been shown to be responsive to stimulation by transforming growth factor (TGF)-β1 or TNF-α. Among these miRNAs, miR-21 demonstrated the greatest increase in UUO kidneys. The enhanced expression of miR-21 was located mainly in distal tubular epithelial cells. miR-21 expression was upregulated in response to treatment with TGF-β1 or TNF-α in human renal tubular epithelial cells in vitro. Furthermore, we found that blocking miR-21 in vivo attenuated UUO-induced renal fibrosis, presumably through diminishing the expression of profibrotic proteins and reducing infiltration of inflammatory macrophages in UUO kidneys. Our data suggest that targeting specific miRNAs could be a novel therapeutic approach to treat renal fibrosis.

Download Full-text

Homeopathic potencies of Arnica montana L. change gene expression in a Tamm-Horsfall protein-1 cell line in vitro model: the role of ethanol as a possible confounder and statistical bias

Journal of Integrative Medicine ◽

10.1016/s2095-4964(17)60346-7 ◽

2017 ◽

Vol 15 (4) ◽

pp. 255-264 ◽

Cited By ~ 4

Author(s):

Salvatore Chirumbolo ◽

Geir Bjørklund

Keyword(s):

Gene Expression ◽

Cell Line ◽

In Vitro Model ◽

Statistical Bias ◽

Arnica Montana ◽

Vitro Model

Download Full-text

Abstract 445: A Pro-Fibrotic Role of Interleukin-4 in Cardiac Remodeling and Dysfunction

Hypertension ◽

10.1161/hyp.64.suppl_1.445 ◽

2014 ◽

Vol 64 (suppl_1) ◽

Author(s):

Hongmei Peng ◽

Oscar Carretero ◽

Xiao-Ping Yang ◽

Pablo Nakagawa ◽

Jiang Xu ◽

...

Keyword(s):

Cardiac Fibrosis ◽

Cardiac Fibroblasts ◽

Interleukin 4 ◽

Ang Ii ◽

Collagen Production ◽

And Function ◽

First Time ◽

Mini Pump

Elevated interleukin-4 (IL-4) levels are positively related to cardiac fibrosis in heart failure and hypertension. Using Balb/c exhibiting high circulating IL-4, Balb/c- Il4 tm2Nnt (IL-4 knockout with Balb/c background, IL-4 -/- ) and C57BL/6 mice, as well as cultured cardiac fibroblasts (CFs), we hypothesized that 1) high levels of IL-4 result in cardiac fibrosis, making the heart susceptible to angiotensin II (Ang II)-induced damage, and 2) IL-4 potently stimulates collagen production by CFs. Each strain (9- to 12-week old male) received vehicle or Ang II (1.4 mg/kg/day, s.c. via osmotic mini-pump) for 8 weeks. Cardiac fibrosis and function were determined by histology and echocardiography, respectively. Compared to C57BL/6, Balb/c mice had doubled interstitial collagen in the heart, enlarged left ventricle and decreased cardiac function along with elevated cardiac IL-4 protein (1.00±0.08 in C57BL/6 vs 2.61±0.46 in Balb/c, p <0.05); all those changes were significantly attenuated in IL-4 -/- (Table 1). Ang II further deteriorated cardiac fibrosis and dysfunction in Balb/c; these detrimental effects were attenuated in IL-4 -/- , although the three strains had a similar level of hypertension. In vitro study revealed that IL-4Rα was constitutively expressed in CFs (Western blot), and IL-4 potently stimulated collagen production by CFs (hydroxproline assay, from 18.89±0.85 to 38.81±3.61 μg/mg at 10 ng/ml, p <0.01). Our study demonstrates for the first time that IL-4, as a potent pro-fibrotic cytokine in the heart, contributes to cardiac fibrotic remodeling and dysfunction. Thus IL-4 may be a potential therapeutic target for cardiac fibrosis and dysfunction.

Download Full-text

HIF-1α contributes to Ang II-induced inflammatory cytokine production in podocytes

BMC Pharmacology and Toxicology ◽

10.1186/s40360-019-0340-8 ◽

2019 ◽

Vol 20 (1) ◽

Cited By ~ 4

Author(s):

Hao Huang ◽

Yanqin Fan ◽

Zhao Gao ◽

Wei Wang ◽

Ning Shao ◽

...

Keyword(s):

Hypoxia Inducible Factor ◽

Renin Angiotensin System ◽

Inflammatory Factors ◽

Ang Ii ◽

Angiotensin System ◽

Inflammatory Injury ◽

Tnf Α

Abstract Background Studies have indicated that changed expression of hypoxia-inducible factor-1α (HIF-1α) in epithelial cells from the kidney could affect the renal function in chronic kidney disease (CKD). As Angiotensin II (Ang II) is a critical active effector in the renin-angiotensin system (RAS) and was proved to be closely related to the inflammatory injury. Meanwhile, researchers found that Ang II could alter the expression of HIF-1α in the kidney. However, whether HIF-1α is involved in mediating Ang II-induced inflammatory injury in podocytes is not clear. Methods Ang II perfusion animal model were established to assess the potential role of HIF-1α in renal injury in vivo. Ang II stimulated podocytes to observe the corresponding between HIF-1α and inflammatory factors in vitro. Results The expression of inflammatory cytokines such as MCP-1 and TNF-α was increased in the glomeruli from rats treated with Ang II infusion compared with control rats. Increased HIF-1α expression in the glomeruli was also observed in Ang II-infused rats. In vitro, Ang II upregulated the expression of HIF-1α in podocytes. Furthermore, knockdown of HIF-1α by siRNA decreased the expression of MCP-1 and TNF-α. Moreover, HIF-1α siRNA significantly diminished the Ang II-induced overexpression of HIF-1α. Conclusion Collectively, our results suggest that HIF-1α participates in the inflammatory response process caused by Ang II and that downregulation of HIF-1α may be able to partially protect or reverse inflammatory injury in podocytes.

Download Full-text

Norepinephrine and ANG II stimulate secretion of TGF-beta by neonatal rat cardiac fibroblasts in vitro

AJP Cell Physiology ◽

10.1152/ajpcell.1995.268.4.c910 ◽

1995 ◽

Vol 268 (4) ◽

pp. C910-C917 ◽

Cited By ~ 60

Author(s):

S. A. Fisher ◽

M. Absher

Keyword(s):

Transforming Growth Factor Beta ◽

Transforming Growth Factor ◽

Cardiac Fibroblasts ◽

Neonatal Rat ◽

Heart Cells ◽

Ang Ii ◽

Tgf Beta ◽

Hypertrophic Growth ◽

Beta 2

Transforming growth factor-beta (TGF-beta) is a ubiquitous growth-regulating protein that is capable of influencing the growth and function of heart cells in vitro. To better understand the role TGF-beta might play as a paracrine mediator of cardiac hypertrophy, the expression, secretion, and growth effects of TGF-beta were examined. Neonatal cardiac fibroblasts in vitro secreted latent TGF-beta 1 and TGF-beta 2 as high as 15 ng/10(6) cells. Angiotensin II (ANG II) and norepinephrine (NE) each augmented up to threefold the expression and secretion of latent TGF-beta 1 and TGF-beta 2 and also induced a shift in isoform predominance from beta 1 to beta 2. Each agent individually produced hypertrophic growth of neonatal cardiocytes and hyperplastic growth of cardiac fibroblasts. Paradoxically, the combination of NE and ANG II at intermediate and high concentrations resulted in less TGF-beta secretion (compared with either agent alone) and in hypertrophic growth of fibroblasts. These results suggest that the growth-promoting effects of ANG II and NE may in part be mediated via a paracrine stimulation of TGF-beta secretion.

Download Full-text