Sodium tanshinone IIA sulfonate attenuates angiotensin II-induced collagen type I expression in cardiac fibroblasts in vitro

Fibronectin is an extracellular matrix glycoprotein with a regulatory role in fundamental cellular processes. Recent reports on the cardioprotective effect of fibronectin inhibition in a setting of myocardial injury suggest a role for fibronectin in cardiac fibroblast function, which remains largely unexplored. This study probed the molecular basis and functional implications of fibronectin gene expression in cardiac fibroblasts exposed to Angiotensin II, a potent pro-fibrotic factor in the myocardium. Using gene knockdown and over-expression approaches, western blotting and promoter pull-down assay, we show that collagen type I-activated Discoidin Domain Receptor 2 (DDR2) mediates Angiotensin II-stimulated transcriptional up-regulation of fibronectin expression by Yes-activated Protein in cardiac fibroblasts. Further, siRNA-mediated fibronectin knockdown attenuated Angiotensin II-dependent expression of anti-apoptotic cIAP2 and promoted cell death under oxidative stress. Fibronectin was also found to mediate Angiotensin II-stimulated collagen type I expression. Importantly, an obligate role for fibronectin was observed in Angiotensin II-stimulated expression of its receptor, AT1R, which would link ECM signalling and Angiotensin II signalling in cardiac fibroblasts. Moreover, the regulatory role of DDR2-dependent fibronectin expression in Ang II-stimulated cIAP2, collagen type I and AT1R expression was mediated by Integrin-β1-integrin-linked kinase signalling. The pro-survival role of fibronectin in cardiac fibroblasts and its regulatory role in collagen and AT1R expression, downstream of DDR2, could be critical determinants of cardiac fibroblast-mediated wound healing following myocardial injury. Our findings point to a complex mechanism of regulation of cardiac fibroblast function involving two major extracellular matrix proteins, collagen type I and fibronectin, and their receptors, DDR2 and Integrin-β1.

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Beneficial Effects of Mineralocorticoid Receptor Antagonism on Myocardial Fibrosis in an Experimental Model of the Myxomatous Degeneration of the Mitral Valve

International Journal of Molecular Sciences ◽

10.3390/ijms21155372 ◽

2020 ◽

Vol 21 (15) ◽

pp. 5372

Author(s):

Jaime Ibarrola ◽

Mattie Garaikoetxea ◽

Amaia Garcia-Peña ◽

Lara Matilla ◽

Eva Jover ◽

...

Keyword(s):

Mitral Valve ◽

Myocardial Fibrosis ◽

Mineralocorticoid Receptor ◽

Collagen Type ◽

Cardiac Fibroblasts ◽

Collagen Type I ◽

Type I ◽

Myxomatous Degeneration ◽

Human Cardiac Fibroblasts

Mitral valve prolapse (MVP) patients develop myocardial fibrosis that is not solely explained by volume overload, but the pathophysiology has not been defined. Mineralocorticoid receptor antagonists (MRAs) improve cardiac function by decreasing cardiac fibrosis in other heart diseases. We examined the role of MRA in myocardial fibrosis associated with myxomatous degeneration of the mitral valve. Myocardial fibrosis has been analyzed in a mouse model of mitral valve myxomatous degeneration generated by pharmacological treatment with Nordexfenfluramine (NDF) in the presence of the MRA spironolactone. In vitro, adult human cardiac fibroblasts were treated with NDF and spironolactone. In an experimental mouse, MRA treatment reduced interstitial/perivascular fibrosis and collagen type I deposition. MRA administration blunted NDF-induced cardiac expression of vimentin and the profibrotic molecules galectin-3/cardiotrophin-1. In parallel, MRA blocked the increase in cardiac non-fibrillar proteins such as fibronectin, aggrecan, decorin, lumican and syndecan-4. The following effects are blocked by MRA: in vitro, in adult human cardiac fibroblasts, NDF-treatment-induced myofibroblast activation, collagen type I and proteoglycans secretion. Our findings demonstrate, for the first time, the contribution of the mineralocorticoid receptor (MR) to the development of myocardial fibrosis associated with mitral valve myxomatous degeneration. MRA could be a therapeutic approach to reduce myocardial fibrosis associated with MVP.

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Angiotensin II stimulates collagen synthesis and expression of collagen type I gene in adult rat cardiac fibroblasts

Matrix Biology ◽

10.1016/0945-053x(94)90088-4 ◽

1994 ◽

Vol 14 (5) ◽

pp. 378

Author(s):

T. Liu ◽

J. Kyle ◽

S.A. Jimenez ◽

R.I. Bashey

Keyword(s):

Angiotensin Ii ◽

Collagen Synthesis ◽

Collagen Type ◽

Cardiac Fibroblasts ◽

Collagen Type I ◽

Type I ◽

Adult Rat ◽

I Gene

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Angiotensin II Regulation of Collagen Type I Expression in Cardiac Fibroblasts

Hypertension ◽

10.1161/01.hyp.0000144400.49062.6b ◽

2004 ◽

Vol 44 (5) ◽

pp. 655-661 ◽

Cited By ~ 104

Author(s):

Kui Chen ◽

Jiawei Chen ◽

Dayuan Li ◽

Xingjian Zhang ◽

Jawahar L. Mehta

Keyword(s):

Angiotensin Ii ◽

Collagen Type ◽

Cardiac Fibroblasts ◽

Collagen Type I ◽

Type I

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Marked attenuation of production of collagen type I from cardiac fibroblasts by dehydroepiandrosterone

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00370.2004 ◽

2005 ◽

Vol 288 (6) ◽

pp. E1222-E1228 ◽

Cited By ~ 20

Author(s):

Tomoyuki Iwasaki ◽

Koji Mukasa ◽

Masato Yoneda ◽

Satoshi Ito ◽

Yoshihiko Yamada ◽

...

Keyword(s):

Mrna Expression ◽

Collagen Type ◽

Cardiac Fibroblasts ◽

Collagen Type I ◽

Protein Accumulation ◽

Transcriptional Level ◽

Type I ◽

Procollagen Type ◽

Procollagen Type I

Dehydroepiandrosterone (DHEA) is a type of adrenal steroid. The concentrations of DHEA and its sulfate (DHEA-S) in serum reach a peak between the ages of 25 and 30 yr and thereafter decline steadily. It was reported that DHEA-S concentration in humans is inversely related to death from cardiovascular diseases. In this study, we examined the effects of DHEA on regulation of collagen mRNA and collagen synthesis in cultured cardiac fibroblasts. Treatment with DHEA (10−6 M) resulted in a significant decrease in procollagen type I mRNA expression compared with controls. This was accompanied by a significant decrease in procollagen type I protein accumulation in the medium and also a significant decrease in procollagen type I protein synthesis in the cellular matrix. Furthermore, to confirm in vitro results, we administered DHEA to Sprague-Dawley rats, which were treated with angiotensin II for 8 wk to induce cardiac damage. Procollagen type I mRNA expression was significantly decreased and cardiac fibrosis significantly inhibited in DHEA-treated rat hearts without lowering the systolic blood pressure. These results strongly indicate that DHEA can directly attenuate collagen type I synthesis at the transcriptional level in vivo and in vitro in cardiac fibroblasts.

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Discoidin Domain Receptor 2 Regulates AT1R Expression in Angiotensin II-Stimulated Cardiac Fibroblasts via Fibronectin-Dependent Integrin-β1 Signaling

International Journal of Molecular Sciences ◽

10.3390/ijms22179343 ◽

2021 ◽

Vol 22 (17) ◽

pp. 9343

Author(s):

Allen Sam Titus ◽

Harikrishnan Venugopal ◽

Mereena George Ushakumary ◽

Mingyi Wang ◽

Randy T. Cowling ◽

...

Keyword(s):

Angiotensin Ii ◽

Collagen Type ◽

Cardiac Fibroblasts ◽

Collagen Type I ◽

Cardiac Fibroblast ◽

Type I ◽

Integrin Β1 ◽

Discoidin Domain Receptor ◽

Discoidin Domain Receptor 2

This study probed the largely unexplored regulation and role of fibronectin in Angiotensin II-stimulated cardiac fibroblasts. Using gene knockdown and overexpression approaches, Western blotting, and promoter pull-down assay, we show that collagen type I-activated Discoidin Domain Receptor 2 (DDR2) mediates Angiotensin II-dependent transcriptional upregulation of fibronectin by Yes-activated Protein in cardiac fibroblasts. Furthermore, siRNA-mediated fibronectin knockdown attenuated Angiotensin II-stimulated expression of collagen type I and anti-apoptotic cIAP2, and enhanced cardiac fibroblast susceptibility to apoptosis. Importantly, an obligate role for fibronectin was observed in Angiotensin II-stimulated expression of AT1R, the Angiotensin II receptor, which would link extracellular matrix (ECM) signaling and Angiotensin II signaling in cardiac fibroblasts. The role of fibronectin in Angiotensin II-stimulated cIAP2, collagen type I, and AT1R expression was mediated by Integrin-β1-integrin-linked kinase signaling. In vivo, we observed modestly reduced basal levels of AT1R in DDR2-null mouse myocardium, which were associated with the previously reported reduction in myocardial Integrin-β1 levels. The role of fibronectin, downstream of DDR2, could be a critical determinant of cardiac fibroblast-mediated wound healing following myocardial injury. In summary, our findings suggest a complex mechanism of regulation of cardiac fibroblast function involving two major ECM proteins, collagen type I and fibronectin, and their receptors, DDR2 and Integrin-β1.

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Relative rates of biosynthesis of collagen type I, type V and type VI in calf cornea

Biochemical Journal ◽

10.1042/bj2740615 ◽

1991 ◽

Vol 274 (2) ◽

pp. 615-617 ◽

Cited By ~ 32

Author(s):

P Kern ◽

M Menasche ◽

L Robert

Keyword(s):

Type I Collagen ◽

Collagen Type ◽

Corneal Stroma ◽

Collagen Type I ◽

Type I ◽

Type Vi Collagen ◽

Sds Page ◽

Proline Incorporation ◽

Type V

The biosynthesis of type I, type V and type VI collagens was studied by incubation of calf corneas in vitro with [3H]proline as a marker. Pepsin-solubilized collagen types were isolated by salt fractionation and quantified by SDS/PAGE. Expressed as proportions of the total hydroxyproline solubilized, corneal stroma comprised 75% type I, 8% type V and 17% type VI collagen. The rates of [3H]proline incorporation, linear up to 24 h for each collagen type, were highest for type VI collagen and lowest for type I collagen. From pulse-chase experiments, the calculated apparent half-lives for types I, V and VI collagens were 36 h, 10 h and 6 h respectively.

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Plant Recombinant Human Collagen Type I Hydrogels for Corneal Regeneration

Regenerative Engineering and Translational Medicine ◽

10.1007/s40883-021-00220-3 ◽

2021 ◽

Author(s):

Michel Haagdorens ◽

Elle Edin ◽

Per Fagerholm ◽

Marc Groleau ◽

Zvi Shtein ◽

...

Keyword(s):

Collagen Type ◽

Collagen Type I ◽

Collagen Fibrils ◽

Type I ◽

Safe Alternative ◽

Human Donor ◽

Human Collagen ◽

Donor Corneas

Abstract Purpose To determine feasibility of plant-derived recombinant human collagen type I (RHCI) for use in corneal regenerative implants Methods RHCI was crosslinked with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to form hydrogels. Application of shear force to liquid crystalline RHCI aligned the collagen fibrils. Both aligned and random hydrogels were evaluated for mechanical and optical properties, as well as in vitro biocompatibility. Further evaluation was performed in vivo by subcutaneous implantation in rats and corneal implantation in Göttingen minipigs. Results Spontaneous crosslinking of randomly aligned RHCI (rRHCI) formed robust, transparent hydrogels that were sufficient for implantation. Aligning the RHCI (aRHCI) resulted in thicker collagen fibrils forming an opaque hydrogel with insufficient transverse mechanical strength for surgical manipulation. rRHCI showed minimal inflammation when implanted subcutaneously in rats. The corneal implants in minipigs showed that rRHCI hydrogels promoted regeneration of corneal epithelium, stroma, and nerves; some myofibroblasts were seen in the regenerated neo-corneas. Conclusion Plant-derived RHCI was used to fabricate a hydrogel that is transparent, mechanically stable, and biocompatible when grafted as corneal implants in minipigs. Plant-derived collagen is determined to be a safe alternative to allografts, animal collagens, or yeast-derived recombinant human collagen for tissue engineering applications. The main advantage is that unlike donor corneas or yeast-produced collagen, the RHCI supply is potentially unlimited due to the high yields of this production method. Lay Summary A severe shortage of human-donor corneas for transplantation has led scientists to develop synthetic alternatives. Here, recombinant human collagen type I made of tobacco plants through genetic engineering was tested for use in making corneal implants. We made strong, transparent hydrogels that were tested by implanting subcutaneously in rats and in the corneas of minipigs. We showed that the plant collagen was biocompatible and was able to stably regenerate the corneas of minipigs comparable to yeast-produced recombinant collagen that we previously tested in clinical trials. The advantage of the plant collagen is that the supply is potentially limitless.

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