Relaxin Modulates Cardiac Fibroblast Proliferation, Differentiation, and Collagen Production and Reverses Cardiac Fibrosis in Vivo

In recent years, Platelet Rich Plasma (PRP) and Adipose-Derived Stem Cells (ADSCs) have been used separately for many clinical applications, especially skin rejuvenation. A combined injection of PRP and ADSCs could therefore be used to treat skin wrinkles. However, there are controversies and reports with conflicting results regarding the efficacy of this treatment. The authors aimed to determine the anti-wrinkle and skin rejuvenation mechanism of combined PRP and ADSCs treatment. The effects of PRP and ADSCs isolated from the same consenting donors were evaluated using in vitro and in vivo models. The in vitro effects of PRP and ADSCs on dermal fibroblast proliferation, collagen production, and inhibition of Matrix Metalloproteinase-1 (MMP-1) production were investigated using a co-culture model. Fibroblasts and ADSCs were cultured within the same dish, but in two separate cavities (using an insert plate), in the presence of the same PRP-supplemented medium. In vivo, the authors evaluated the effects of combined PRP and ADSCs on skin histochemistry, including changes in the dermal layer and collagen production in photo-aged skin (mice). They also determined the survival and differentiation of grafted ADSCs. The results show that combined PRP and ADSCs strongly stimulate in vitro fibroblast proliferation, collagen production, and inhibition of MMP-1 synthesis. Intra-dermal co-injection of PRP and ADSCs was observed to stimulate increased dermal layer thickness and collagen production compared with the untreated group. These results indicate that a combined PRP and ADSC injection can reduce wrinkles more effectively than either PRP or ADSC alone, and provide insight into the clinical use of PRP combined with ADSCs for dermal applications, particularly skin rejuvenation.

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Dual Role of Triptolide in Interrupting the NLRP3 Inflammasome Pathway to Attenuate Cardiac Fibrosis

International Journal of Molecular Sciences ◽

10.3390/ijms20020360 ◽

2019 ◽

Vol 20 (2) ◽

pp. 360 ◽

Cited By ~ 15

Author(s):

Xi-Chun Pan ◽

Ya Liu ◽

Yan-Yan Cen ◽

Ya-Lan Xiong ◽

Jing-Mei Li ◽

...

Keyword(s):

Nlrp3 Inflammasome ◽

Cardiac Fibrosis ◽

Cardiac Fibroblasts ◽

Dual Role ◽

Receptor Protein ◽

Inflammasome Activation ◽

Collagen Production ◽

Antifibrotic Effect ◽

Myeloid Differentiation Factor

In a previous paper, we reported that triptolide (TP), a commonly used immunomodulator, could attenuate cardiac hypertrophy. This present study aimed to further explore the inhibition of cardiac fibrosis by TP and the possible mechanism from the perspective of the NOD-like receptor protein 3 (NLRP3) inflammasome. Hematoxylin-eosin and Masson’s staining, immunohistochemistry, and immunofluorescence were performed to observe cardiac fibrotic changes in mice and mouse cardiac fibroblasts (CFs). The Western blot, colocalization, and immunoprecipitation were applied to detect protein expression and interactions. Results suggested that TP dose-dependently inhibited cardiac fibrosis induced by isoproterenol and collagen production of CFs induced by angiotensin II. TP exhibited an antifibrotic effect via inhibiting activation of the NLRP3 inflammasome, which sequentially decreased IL-1β maturation, myeloid differentiation factor 88 (MyD88)-related phosphorylation of c-Jun N-terminal kinase (JNK), extracellular regulated protein kinase 1/2 (ERK1/2), and TGF-β1/Smad signaling, and ultimately resulted in less collagen production. Moreover, TP showed no antifibrotic effect in Nlrp3-knockout CFs. Notably, TP inhibited the expression of NLRP3 and apoptosis-associated speck-like proteins containing a caspase recruitment domain (ASC) as well as inflammasome assembly, by interrupting the NLRP3-ASC interaction to inhibit inflammasome activation. Finally, TP indeed inhibited the NLRP3-TGFβ1-Smad pathway in vivo. Conclusively, TP was found to play a dual role in interrupting the activation of the NLRP3 inflammasome to attenuate cardiac fibrosis.

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Can Activated Platelet Rich Plasma Combined with Adipose-Derived Stem Cells Be Used to Treat Skin Wrinkles?

Bioinformatics ◽

10.4018/978-1-4666-3604-0.ch049 ◽

2013 ◽

pp. 920-936

Author(s):

Phuc Van Pham ◽

Loan Thi-Tung Dang ◽

Nhung Hai Truong ◽

Ngoc Kim Phan

Keyword(s):

Stem Cells ◽

Platelet Rich Plasma ◽

Adipose Derived Stem Cells ◽

Fibroblast Proliferation ◽

Skin Rejuvenation ◽

Collagen Production ◽

Dermal Layer ◽

Skin Wrinkles

In recent years, Platelet Rich Plasma (PRP) and Adipose-Derived Stem Cells (ADSCs) have been used separately for many clinical applications, especially skin rejuvenation. A combined injection of PRP and ADSCs could therefore be used to treat skin wrinkles. However, there are controversies and reports with conflicting results regarding the efficacy of this treatment. The authors aimed to determine the anti-wrinkle and skin rejuvenation mechanism of combined PRP and ADSCs treatment. The effects of PRP and ADSCs isolated from the same consenting donors were evaluated using in vitro and in vivo models. The in vitro effects of PRP and ADSCs on dermal fibroblast proliferation, collagen production, and inhibition of Matrix Metalloproteinase-1 (MMP-1) production were investigated using a co-culture model. Fibroblasts and ADSCs were cultured within the same dish, but in two separate cavities (using an insert plate), in the presence of the same PRP-supplemented medium. In vivo, the authors evaluated the effects of combined PRP and ADSCs on skin histochemistry, including changes in the dermal layer and collagen production in photo-aged skin (mice). They also determined the survival and differentiation of grafted ADSCs. The results show that combined PRP and ADSCs strongly stimulate in vitro fibroblast proliferation, collagen production, and inhibition of MMP-1 synthesis. Intra-dermal co-injection of PRP and ADSCs was observed to stimulate increased dermal layer thickness and collagen production compared with the untreated group. These results indicate that a combined PRP and ADSC injection can reduce wrinkles more effectively than either PRP or ADSC alone, and provide insight into the clinical use of PRP combined with ADSCs for dermal applications, particularly skin rejuvenation.

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Abstract 77: MiR-125b Regulates Myofibroblast Transition and Cardiac Fibrosis

Circulation Research ◽

10.1161/res.117.suppl_1.77 ◽

2015 ◽

Vol 117 (suppl_1) ◽

Author(s):

Varun Nagpal ◽

Rahul Rai ◽

Aaron T Place ◽

Asish K Ghosh ◽

Douglas E Vaughan

Keyword(s):

Cardiac Fibrosis ◽

Transforming Growth Factor ◽

Primary Cultures ◽

Locked Nucleic Acid ◽

Mouse Heart ◽

Ang Ii ◽

Fibroblast Proliferation ◽

Systemic Delivery ◽

Critical Determinant

Transforming growth factor-β (TGF-β)-induced fibroblast-to-myofibroblast transition (FMT) is a critical determinant of cardiac fibrosis. However, the contribution of microRNAs leading to TGF-β-induced FMT and cardiac fibrosis are not well-understood. Our results elucidate that blocking the canonical TGF-β pathway protects from FMT in primary cultures of human cardiac fibroblasts and that miR-125b is significantly upregulated during cardiac FMT. Furthermore, we observed significant upregulation of miR-125b in fibrotic human myocardium and two murine models of cardiac fibrosis. Importantly, we discovered that miR-125b is sufficient to induce cardiac FMT. In contrast, the knockdown of miR-125b using an antagomir approach attenuated TGF-β-induced FMT. In silico analysis and biochemical analysis revealed that miR-125b directly targets multiple anti-fibrotic mediators including p53 and apelin. In addition, miR-125b also plays a potent role in the regulation of fibroblast proliferation, an important cause of cardiac fibrosis. Finally, miR-125b was successfully inhibited in vivo by the systemic delivery of locked nucleic acid (LNA) targeted against miR-125b both in the presence and absence of Angiotensin II (Ang II). These results demonstrated that LNA-125b protected against Ang II-induced proliferation and fibrosis in the mouse heart in vivo. We conclude that TGF-β-induced miR-125b is an important regulator of both fibroblast proliferation and FMT, and miR-125b inhibits key anti-fibrotic mediators to promote cardiac fibrosis. We propose that miR-125b may serve as a novel therapeutic target for the preventative therapy for fibrosis.

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Abstract 415: Asporin Contributes to Reactive Interstitial Cardiac Fibrosis by Regulating Cardiac Fibroblast Activation

Circulation Research ◽

10.1161/res.127.suppl_1.415 ◽

2020 ◽

Vol 127 (Suppl_1) ◽

Author(s):

Lejla Medzikovic ◽

Laila Aryan ◽

Gregoire Ruffenach ◽

Mansoureh Eghbali

Keyword(s):

Cardiac Fibrosis ◽

Ischemia Reperfusion Injury ◽

Interstitial Fibrosis ◽

Smooth Muscle Actin ◽

Ischemia Reperfusion ◽

Bioinformatic Analysis ◽

Gene Signature ◽

Cardiac Fibroblast ◽

Collagen Production

Cardiac fibrosis critically contributes to heart failure progression. Depending on the pathological insult, cardiac fibrosis either replaces necrotic cardiomyocytes or is reactive to cardiac fibroblast (CF) activation. The extracellular matrix (ECM) consists of various proteins and the role of fibrillar collagen has been well studied. However, the role of non-fibrillar ECM proteins in cardiac fibrosis is less clear. To explore the role of ECM in reactive cardiac fibrosis, we performed bioinformatic analysis on online available microarray GEO datasets from hearts of human hypertrophic cardiomyopathy patients and two mouse models of transverse aortic constriction and Angiotensin II (AngII) infusion. We found that 27 differentially expressed genes were common between the three datasets. Among these genes was the small leucine-rich proteoglycan Asporin (ASPN). ASPN was previously shown to be upregulated in the ECM of replacement fibrosis in porcine ischemia/reperfusion injury. However, not much is known about the role of ASPN in reactive interstitial fibrosis. We show that cardiac ASPN expression is enhanced in mice after short- and long-term AngII infusion compared to saline infusion. In resident CF isolated from adult mice, ASPN expression is upregulated by both AngII and TGF-β stimulation. Here, ASPN expression correlates with a gene signature of activated CFs including periostin ( postn ), α-smooth muscle actin ( acta2 ) and collagens I and III ( col1a1, col3a1 ), and with functional characteristics of activated CFs including proliferation, migration and collagen production. Modulating ASPN via siRNA in mouse resident CFs inhibits postn, acta2, col1a1 and col3a1 expression and total collagen production, indicating repressed CF activation upon ASPN knockdown. Taken together, ASPN may be an attractive novel target against reactive interstitial fibrosis.

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Repurposing mesalazine against cardiac fibrosis in vitro

Naunyn-Schmiedeberg s Archives of Pharmacology ◽

10.1007/s00210-020-01998-9 ◽

2020 ◽

Author(s):

Maximilian Hoffmann ◽

Theresa A. Kant ◽

Ramona Emig ◽

Johanna S. E. Rausch ◽

Manja Newe ◽

...

Keyword(s):

Cardiac Fibrosis ◽

Drug Repurposing ◽

Fibroblast Cell ◽

Cardiac Fibroblast ◽

Cell Stiffness ◽

Fibroblast Proliferation ◽

Time Saving ◽

Solvent Control ◽

Vitro Model

Abstract Cardiovascular diseases are exacerbated and driven by cardiac fibrosis. TGFβ induces fibroblast activation and differentiation into myofibroblasts that secrete excessive extracellular matrix proteins leading to stiffening of the heart, concomitant cardiac dysfunction, and arrhythmias. However, effective pharmacotherapy for preventing or reversing cardiac fibrosis is presently unavailable. Therefore, drug repurposing could be a cost- and time-saving approach to discover antifibrotic interventions. The aim of this study was to investigate the antifibrotic potential of mesalazine in a cardiac fibroblast stress model. TGFβ was used to induce a profibrotic phenotype in a human cardiac fibroblast cell line. After induction, cells were treated with mesalazine or solvent control. Fibroblast proliferation, key fibrosis protein expression, extracellular collagen deposition, and mechanical properties were subsequently determined. In response to TGFβ treatment, fibroblasts underwent a profound phenoconversion towards myofibroblasts, determined by the expression of fibrillary αSMA. Mesalazine reduced differentiation nearly by half and diminished fibroblast proliferation by a third. Additionally, TGFβ led to increased cell stiffness and adhesion, which were reversed by mesalazine treatment. Collagen 1 expression and deposition—key drivers of fibrosis—were significantly increased upon TGFβ stimulation and reduced to control levels by mesalazine. SMAD2/3 and ERK1/2 phosphorylation, along with reduced nuclear NFκB translocation, were identified as potential modes of action. The current study provides experimental pre-clinical evidence for antifibrotic effects of mesalazine in an in vitro model of cardiac fibrosis. Furthermore, it sheds light on possible mechanisms of action and suggests further investigation in experimental and clinical settings.

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Intermittent hypoxia mediated by TSP1 dependent on STAT3 induces cardiac fibroblast activation and cardiac fibrosis

eLife ◽

10.7554/elife.49923 ◽

2020 ◽

Vol 9 ◽

Cited By ~ 5

Author(s):

Qiankun Bao ◽

Bangying Zhang ◽

Ya Suo ◽

Chen Liu ◽

Qian Yang ◽

...

Keyword(s):

Intermittent Hypoxia ◽

Cardiac Fibrosis ◽

Synergistic Effects ◽

Cardiac Fibroblasts ◽

Cardiac Fibroblast ◽

Ang Ii ◽

Fibroblast Activation ◽

Obstructive Sleep

Intermittent hypoxia (IH) is the predominant pathophysiological disturbance in obstructive sleep apnea (OSA), known to be independently associated with cardiovascular diseases. However, the effect of IH on cardiac fibrosis and molecular events involved in this process are unclear. Here, we tested IH in angiotensin II (Ang II)-induced cardiac fibrosis and signaling linked to fibroblast activation. IH triggered cardiac fibrosis and aggravated Ang II-induced cardiac dysfunction in mice. Plasma thrombospondin-1 (TSP1) content was upregulated in both IH-exposed mice and OSA patients. Moreover, both in vivo and in vitro results showed IH-induced cardiac fibroblast activation and increased TSP1 expression in cardiac fibroblasts. Mechanistically, phosphorylation of STAT3 at Tyr705 mediated the IH-induced TSP1 expression and fibroblast activation. Finally, STAT3 inhibitor S3I-201 or AAV9 carrying a periostin promoter driving the expression of shRNA targeting Stat3 significantly attenuated the synergistic effects of IH and Ang II on cardiac fibrosis in mice. This work suggests a potential therapeutic strategy for OSA-related fibrotic heart disease.

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Abstract 167: Mir-195-3p/-5p Decrease Cardiac Fibroblast Proliferation and the Transdifferentiation into Myofibroblasts

Circulation Research ◽

10.1161/res.117.suppl_1.167 ◽

2015 ◽

Vol 117 (suppl_1) ◽

Author(s):

Shutong Shen ◽

Xiuzhi Wang ◽

Dongjie Xu ◽

Lichan Tao ◽

Xiaoting Wu ◽

...

Keyword(s):

Target Gene ◽

Cardiac Fibrosis ◽

Cardiac Fibroblast ◽

Immunofluorescent Staining ◽

Fibroblast Proliferation ◽

Chain Reactions ◽

Protein Levels ◽

Polymerase Chain Reactions ◽

Polymerase Chain

Aims: MicroRNAs (miRNAs, miRs) contribute to many essential physiological and pathological processes including fibrosis. This study aims at investigating the role of miR-195-3p/-5p in cardiac fibroblast proliferation and the transdifferentiation into myofibroblasts. Methods and results: In isolated primary neonatal cardiac fibrobasts (NRCFs), forced expression of miR-195-3p/-5p with agomiRs could attenuate fibrobast proliferation as determined by EdU and Ki67 staining while inhibition of miR-195-3p/-5p with antagomiRs could increase fibrobast proliferation. By quantitative reverse transcription polymerase chain reactions (RT-PCRs) and western blotting (WB), α-SMA (a marker of myofibroblast transdifferentiation) was found to be suppressed in the miR-195-3p/-5p agomiR-treated NRCFs at both mRNA and protein levels, while was increased in the miR-195-3p/-5p antagomiR-treated NRCFs. Moreover, Chek-1 was identified as a target gene of miR-195-3p/-5p responsible for cardiac fibroblast proliferation and the transdifferentiation into myofibroblasts by RT-PCR and WB and immunofluorescent staining. Silencing of Chek-1 attenuates cardiac fibroblast proliferation and the transdifferentiation into myofibroblasts as detected by α-SMA/EDU staining. In addition, Chek-1 mediated the effects of miR-195-3p/-5p in cardiac fibroblast proliferation and the transdifferentiation into myofibroblasts. Conclusion: Therefore, miR-195-3p/-5p might be promising therapeutic targets for cardiac fibrosis.

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MicroRNA‐26a Protects the Heart Against Hypertension‐Induced Myocardial Fibrosis

Journal of the American Heart Association ◽

10.1161/jaha.120.017970 ◽

2020 ◽

Vol 9 (18) ◽

Author(s):

Wenqian Zhang ◽

Qiaozhu Wang ◽

Yanjing Feng ◽

Xuegui Chen ◽

Lijun Yang ◽

...

Keyword(s):

Blood Pressure ◽

Myocardial Fibrosis ◽

Blood Pressure Control ◽

Cardiac Fibroblasts ◽

Pressure Control ◽

Cardiac Fibroblast ◽

Tissue Growth ◽

Fibroblast Proliferation ◽

Spontaneously Hypertensive

Background Hypertensive myocardial fibrosis (MF) is characterized by excessive deposition of extracellular matrix and cardiac fibroblast proliferation, which can lead to heart failure, malignant arrhythmia, and sudden death. In recent years, with the deepening of research, microRNAs have been found to have an important role in blood pressure control and maintaining normal ventricular structure and function. Methods and Results In this study, we first documented the downregulation of microRNA‐26a (miR‐26a) in the plasma and myocardium of spontaneously hypertensive rats; more importantly, miR‐26a–deficient mice showed MF, whereas overexpression of miR‐26a significantly prevented elevated blood pressure and inhibited MF in vivo and angiotensin II‐induced fibrogenesis in cardiac fibroblasts by directly targeting connective tissue growth factor and Smad4. miR‐26a inhibited cardiac fibroblast proliferation by the enhancer of zeste homolog 2/p21 pathway. Conclusions Our study identified a novel role for miR‐26a in blood pressure control and hypertensive MF and provides a possible treatment strategy for miR‐26a to alleviate and reverse hypertensive MF.

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