Pubblicazioni recenti - cardiac fibroblast
-
Endothelial Cell-Specific Prolyl Hydroxylase-2 Deficiency Augments Angiotensin II-Induced Arterial Stiffness and Cardiac Pericyte Recruitment in Mice.
J Am Heart Assoc2024 Jul;():e035769. doi: 10.1161/JAHA.124.035769.
Liu Bo, Zeng Heng, Su Han, Williams Quinesha A, Besanson Jessie, Chen Yingjie, Chen Jian-Xiong,
Abstract
BACKGROUND:
Endothelial prolyl hydroxylase-2 (PHD2) is essential for pulmonary remodeling and hypertension. In the present study, we investigated the role of endothelial PHD2 in angiotensin II-mediated arterial stiffness, pericyte recruitment, and cardiac fibrosis.
METHODS AND RESULTS:
Chondroitin sulfate proteoglycan 4 tracing reporter chondroitin sulfate proteoglycan 4- red fluorescent protein (DsRed) transgenic mice were crossed with PHD2 (PHD2) mice and endothelial-specific knockout of PHD2 (PHD2KO) mice. Transgenic PHD2 (TgPHD2) mice and TgPHD2KO mice were infused with angiotensin II for 4?weeks. Arterial thickness, stiffness, and histological and immunofluorescence of pericytes and fibrosis were measured. Infusion of TgPHD2 mice with angiotensin II resulted in a time-dependent increase in pulse-wave velocity. Angiotensin II-induced pulse-wave velocity was further elevated in the TgPHD2KO mice. TgPHD2KO also reduced coronary flow reserve compared with TgPHD2 mice infused with angiotensin II. Mechanistically, knockout of endothelial PHD2 promoted aortic arginase activity and angiotensin II-induced aortic thickness together with increased transforming growth factor-?1 and ICAM-1/VCAM-1 expression in coronary arteries. TgPHD2 mice infused with angiotensin II for 4?weeks exhibited a significant increase in cardiac fibrosis and hypertrophy, which was further developed in the TgPHD2KO mice. Chondroitin sulfate proteoglycan 4 pericyte was traced by DsRed staining and angiotensin II infusion displayed a significant increase of DsRed pericytes in the heart, as well as a deficiency of endothelial PHD2, which further promoted angiotensin II-induced pericyte increase. DsRed pericytes were costained with fibroblast-specific protein 1 and ?-smooth muscle actin for measuring pericyte-myofibroblast cell transition. The knockout of endothelial PHD2 increased the amount of DsRed/fibroblast-specific protein 1 and DsRed/?-smooth muscle actin cells induced by angiotensin II infusion.
CONCLUSIONS:
Knockout of endothelial PHD2 enhanced angiotensin II-induced cardiac fibrosis by mechanisms involving increasing arterial stiffness and pericyte-myofibroblast cell transitions.
Guarda su PubMed -
The effects of berberine and curcumin on cardiac, lipid profile and fibrosis markers in cyclophosphamide-induced cardiac damage: The role of the TRPM2 channel.
J Biochem Mol Toxicol2024 Aug;38(8):e23783. doi: 10.1002/jbt.23783.
Huyut Zübeyir, Yildizhan Kenan, Alt?nda? Fikret,
Abstract
Cyclophosphamide (CYP) is widely used to treat various types of cancer. In addition to the therapeutic properties of this drug, unfortunately, its side effects are still not fully understood. This study investigated the protective effect of curcumin (CURC) and berberine (BER) on CYP-induced cardiac damage. Thirty-six male rats were equally divided into the control, dimethyl sulfoxide (DMSO), CYP, CYP?+?CURC, CYP?+?BER and CYP?+?BER?+?CURC groups. Troponin-I, Creatine kinase-myocardial band (CK-MB), total cholesterol, triglyceride levels in serum samples, and reactive oxygen species (ROS), poly(ADP-ribose) polymerase-1 (PARP-1), and transient receptor potential melastatin 2 (TRPM2) channel levels in heart tissue were measured using an enzyme-linked immunoassay (ELISA) kit. In addition, histopathological examination and immunohistochemical investigation of the TRPM2 channel, fibroblast specific protein-1 (FSP1), transforming growth factor-beta- 1 (TGF-?1) and ?-smooth muscle actin (?-SMA) expressions were determined in heart tissue. The CYP group's troponin-I, total cholesterol, triglyceride, CK-MB, ROS, PARP-1 and TRPM2 channel levels were higher than in the other groups in the ELISA measurements (p?0.05). In contrast, these parameters in the group treated with CURC and BER together with CYP were lower than in the CYP group (p?0.05). Additionally, CUR and BER reduced CYP-induced pathological damage, TRPM2, FSP1, TGF-?1 and ?-SMA expressions. The data showed that CYP administration can cause cardiac damage by increasing the TRPM2 channel, TGF-?1, FSP1 and ?-SMA expression levels. Therefore, we concluded that CURC and BER administration following CYP application may be used as therapeutic agents to prevent CYP-induced cardiac damage.
© 2024 Wiley Periodicals LLC.
Guarda su PubMed -
Primary Human Cell-Derived Extracellular Matrix from Decellularized Fibroblast Microtissues with Tissue-Dependent Composition and Microstructure.
Cell Mol Bioeng2024 Jun;17(3):189-201. doi: 10.1007/s12195-024-00809-y.
Fonseca Vera C, Van Vivian, Ip Blanche C,
Abstract
PURPOSE:
Human extracellular matrix (ECM) exhibits complex protein composition and architecture depending on tissue and disease state, which remains challenging to reverse engineer. One promising approach is based on cell-secreted ECM from primary human fibroblasts that can be decellularized into acellular biomaterials. However, fibroblasts cultured on rigid culture plastic or biomaterial scaffolds can experience aberrant mechanical cues that perturb the biochemical, mechanical, and the efficiency of ECM production.
METHODS:
Here, we demonstrate a method for preparing decellularized ECM using primary human fibroblasts with tissue and disease-specific features with two case studies: (1) cardiac fibroblasts; (2) lung fibroblasts from healthy or diseased donors. Cells aggregate into engineered microtissues in low adhesion microwells that deposited ECM and can be decellularized. We systematically investigate microtissue morphology, matrix architecture, and mechanical properties, along with transcriptomic and proteomic analysis.
RESULTS:
Microtissues exhibited tissue-specific gene expression and proteomics profiling, with ECM complexity similar to native tissues. Healthy lung microtissues exhibited web-like fibrillar collagen compared to dense patches in healthy heart microtissues. Diseased lung exhibited more disrupted collagen architecture than healthy. Decellularized microtissues had tissue-specific mechanical stiffness that was physiologically relevant. Importantly, decellularized microtissues supported viability and proliferation of human cells.
CONCLUSIONS:
We show that engineered microtissues of primary human fibroblasts seeded in low-adhesion microwells can be decellularized to produce human, tissue and disease-specific ECM. This approach should be widely applicable for generating personalized matrix that recapitulate tissues and disease states, relevant for culturing patient cells ex vivo as well as implantation for therapeutic treatments.
SUPPLEMENTARY INFORMATION:
The online version contains supplementary material available at 10.1007/s12195-024-00809-y.
© The Author(s), under exclusive licence to Biomedical Engineering Society 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
Guarda su PubMed -
A novel Na1.5-dependent feedback mechanism driving glycolytic acidification in breast cancer metastasis.
Oncogene2024 Jul;():. doi: 10.1038/s41388-024-03098-x.
Leslie Theresa K, Tripp Aurelien, James Andrew D, Fraser Scott P, Nelson Michaela, Sajjaboontawee Nattanan, Capatina Alina L, Toss Michael, Fadhil Wakkas, Salvage Samantha C, Garcia Mar Arias, Beykou Melina, Rakha Emad, Speirs Valerie, Bakal Chris, Poulogiannis George, Djamgoz Mustafa B A, Jackson Antony P, Matthews Hugh R, Huang Christopher L-H, Holding Andrew N, Chawla Sangeeta, Brackenbury William J,
Abstract
Solid tumours have abnormally high intracellular [Na]. The activity of various Na channels may underlie this Na accumulation. Voltage-gated Na channels (VGSCs) have been shown to be functionally active in cancer cell lines, where they promote invasion. However, the mechanisms involved, and clinical relevance, are incompletely understood. Here, we show that protein expression of the Na1.5 VGSC subtype strongly correlates with increased metastasis and shortened cancer-specific survival in breast cancer patients. In addition, VGSCs are functionally active in patient-derived breast tumour cells, cell lines, and cancer-associated fibroblasts. Knockdown of Na1.5 in a mouse model of breast cancer suppresses expression of invasion-regulating genes. Na1.5 activity increases ATP demand and glycolysis in breast cancer cells, likely by upregulating activity of the Na/K ATPase, thus promoting H production and extracellular acidification. The pH of murine xenograft tumours is lower at the periphery than in the core, in regions of higher proliferation and lower apoptosis. In turn, acidic extracellular pH elevates persistent Na influx through Na1.5 into breast cancer cells. Together, these findings show positive feedback between extracellular acidification and the movement of Na into cancer cells which can facilitate invasion. These results highlight the clinical significance of Na1.5 activity as a potentiator of breast cancer metastasis and provide further evidence supporting the use of VGSC inhibitors in cancer treatment.
© 2024. The Author(s).
Guarda su PubMed -
The Mechanism of SIRT3 Regulating SRV2-Mediated Mitochondrial Fission of Fibroblasts to Inhibit Myocardial Fibrosis after Acute Myocardial Infarction.
Ann Clin Lab Sci2024 May;54(3):335-346.
Zhou Jia, Chen Yuanyuan, Li Qiang, Wang Guodong, Zhang Gao,
Abstract
OBJECTIVE:
Cardiac dysfunction can result from excessive fibrosis in cardiac fibroblasts (CFs) following an acute myocardial infarction (AMI). SIRT3 has been shown to be associated with numerous cardiovascular diseases. This study aimed to investigate the mechanism by which SIRT3 influences myocardial fibrosis following AMI.
METHODS:
An AMI model was established in rats and echocardiography was used to assess cardiac systolic function. Triphenyl tetrazolium chloride (TTC) and H&E staining were employed to observe the myocardial histopathological status. Masson trichrome staining was used to detect fibrosis, and the changes in expression of fibrosis-related proteins were detected by Western Blot (WB). In this study, we utilized in vitro cell models stimulated by Ang II to investigate the underlying mechanisms. We employed Transwell and CCK-8 assays to detect the function of CFs. Additionally, we used transmission electron microscopy (TEM) to observe the structural morphology of mitochondria, whereas WB was performed to quantify fibrosis-associated proteins and to assay the changes in SIRT3, SRV2, and Drp1.
RESULTS:
We observed a significant decrease in the expression of SIRT3 and an increase in mitochondrial fragmentation in rats with AMI. Additionally, we observed upregulation of fibrosis-associated signature proteins and collagen proteins expression. Through the use of vitro Ang II stimulation we observed a downregulation of SIRT3 expression, an increase in mitochondrial fragmentation, and an increase in the proliferation and migration of CFs. Opposite effects were observed when SIRT3 was overexpressed. Additive mitochondrial division agonists were found to stimulate the proliferation and migration of CFs, however, SIRT3 expression was unchanged. Interference with SRV2 and SIRT3 revealed that SIRT3 effectively prevented the expression of SRV2/Drp1, resulting in the inhibition of mitochondrial division and the suppression of CFs proliferative migration.
CONCLUSION:
In summary, SIRT3 can suppress myocardial fibrosis after acute myocardial infarction by regulating SRV2/Drp1-mediated mitochondrial division.
© 2024 by the Association of Clinical Scientists, Inc.
Guarda su PubMed -
GPR176 promotes fibroblast-to-myofibroblast transition in organ fibrosis progression.
Biochim Biophys Acta Mol Cell Res2024 Jul;1871(7):119798. doi: 10.1016/j.bbamcr.2024.119798.
Okamoto Yasuo, Kitakaze Keisuke, Takenouchi Yasuhiro, Matsui Rena, Koga Daisuke, Miyashima Ryo, Ishimaru Hironobu, Tsuboi Kazuhito,
Abstract
Fibrosis is characterized by excessive deposition of extracellular matrix proteins, particularly collagen, caused by myofibroblasts in response to chronic inflammation. Although G protein-coupled receptors (GPCRs) are among the targets of current antifibrotic drugs, no drug has yet been approved to stop fibrosis progression. Herein, we aimed to identify GPCRs with profibrotic effects. In gene expression analysis of mouse lungs with induced fibrosis, eight GPCRs were identified, showing a >2-fold increase in mRNA expression after fibrosis induction. Among them, we focused on Gpr176 owing to its significant correlation with a myofibroblast marker ?-smooth muscle actin (?SMA), the profibrotic factor transforming growth factor ?1 (TGF?1), and collagen in a human lung gene expression database. Similar to the lung fibrosis model, increased Gpr176 expression was also observed in other organs affected by fibrosis, including the kidney, liver, and heart, suggesting its role in fibrosis across various organs. Furthermore, fibroblasts abundantly expressed Gpr176 compared to alveolar epithelial cells, endothelial cells, and macrophages in the fibrotic lung. GPR176 expression was unaffected by TGF?1 stimulation in rat renal fibroblast NRK-49 cells, whereas knockdown of Gpr176 by siRNA reduced TGF?1-induced expression of ?SMA, fibronectin, and collagen as well as Smad2 phosphorylation. This suggested that Gpr176 regulates fibroblast activation. Consequently, Gpr176 acts in a profibrotic manner, and inhibiting its activity could potentially prevent myofibroblast differentiation and improve fibrosis. Developing a GPR176 inverse agonist or allosteric modulator is a promising therapeutic approach for fibrosis.
Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.
Guarda su PubMed -
3D matrix stiffness modulation unveils cardiac fibroblast phenotypic switching.
Sci Rep2024 Jul;14(1):17015. doi: 17015.
Han Yan, Shao Zehua, Zhang Yuanhao, Zhao Huan, Sun Zirui, Yang Chaokuan, Tang Hao, Han Yu, Gao Chuanyu,
Abstract
This study investigates how dynamic fluctuations in matrix stiffness affect the behavior of cardiac fibroblasts (CFs) within a three-dimensional (3D) hydrogel environment. Using hybrid hydrogels with tunable stiffness, we created an in vitro model to mimic the varying stiffness of the cardiac microenvironment. By manipulating hydrogel stiffness, we examined CF responses, particularly the expression of ?-smooth muscle actin (?-SMA), a marker of myofibroblast differentiation. Our findings reveal that increased matrix stiffness promotes the differentiation of CFs into myofibroblasts, while matrix softening reverses this process. Additionally, we identified the role of focal adhesions and integrin ?1 in mediating stiffness-induced phenotypic switching. This study provides significant insights into the mechanobiology of cardiac fibrosis and suggests that modulating matrix stiffness could be a potential therapeutic strategy for treating cardiovascular diseases.
© 2024. The Author(s).
Guarda su PubMed -
CircRNA_012164/MicroRNA-9-5p axis mediates cardiac fibrosis in diabetic cardiomyopathy.
PLoS One2024 ;19(7):e0302772. doi: e0302772.
Wang Honglin, Wang Eric Zi Rui, Feng Biao, Chakrabarti Subrata,
Abstract
Noncoding RNAs play a part in many chronic diseases and interact with each other to regulate gene expression. MicroRNA-9-5p (miR9) has been thought to be a potential inhibitor of diabetic cardiomyopathy. Here we examined the role of miR9 in regulating cardiac fibrosis in the context of diabetic cardiomyopathy. We further expanded our studies through investigation of a regulatory circularRNA, circRNA_012164, on the action of miR9. We showed at both the in vivo and in vitro level that glucose induced downregulation of miR9 and upregulation of circRNA_012164 resulted in the subsequent upregulation of downstream fibrotic genes. Further, knockdown of circRNA_012164 shows protective effects in cardiac endothelial cells and reverses increased transcription of genes associated with fibrosis and fibroblast proliferation through a regulatory axis with miR9. This study presents a novel regulatory axis involving noncoding RNA that is evidently important in the development of cardiac fibrosis in diabetic cardiomyopathy.
Copyright: © 2024 Wang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Guarda su PubMed -
Knockout of C1q/tumor necrosis factor-related protein-9 aggravates cardiac fibrosis in diabetic mice by regulating YAP-mediated autophagy.
Front Pharmacol2024 ;15():1407883. doi: 1407883.
Ruan Shiyan, Li Jun, Lei Shengyun, Zhang Shaomeng, Xu Dan, Zuo Anju, Li Linxi, Guo Yuan,
Abstract
INTRODUCTION:
Diabetic cardiomyopathy (DCM) is predominantly distinguished by impairment in ventricular function and myocardial fibrosis. Previous studies revealed the cardioprotective properties of C1q/tumor necrosis factor-related protein 9 (CTRP9). However, whether CTRP9 affects diabetic myocardial fibrosis and its underlying mechanisms remains unclear.
METHODS:
We developed a type 1 diabetes (T1DM) model in CTRP9-KO mice via streptozotocin (STZ) induction to examine cardiac function, histopathology, fibrosis extent, Yes-associated protein (YAP) expression, and the expression of markers for autophagy such LC3-II and p62. Additionally, we analyzed the direct impact of CTRP9 on high glucose (HG)-induced transdifferentiation, autophagic activity, and YAP protein levels in cardiac fibroblasts.
RESULTS:
In diabetic mice, CTRP9 expression was decreased in the heart. The absence of CTRP9 aggravated cardiac dysfunction and fibrosis in mice with diabetes, alongside increased YAP expression and impaired autophagy. , HG induced the activation of myocardial fibroblasts, which demonstrated elevated cell proliferation, collagen production, and ?-smooth muscle actin (?-SMA) expression. CTRP9 countered these adverse effects by restoring autophagy and reducing YAP protein levels in cardiac fibroblasts. Notably, the protective effects of CTRP9 were negated by the inhibition of autophagy with chloroquine (CQ) or by YAP overexpression through plasmid intervention. Notably, the protective effect of CTRP9 was negated by inhibition of autophagy caused by chloroquine (CQ) or plasmid intervention with YAP overexpression.
DISCUSSION:
Our findings suggest that CTRP9 can enhance cardiac function and mitigate cardiac remodeling in DCM through the regulation of YAP-mediated autophagy. CTRP9 holds promise as a potential candidate for pharmacotherapy in managing diabetic cardiac fibrosis.
Copyright © 2024 Ruan, Li, Lei, Zhang, Xu, Zuo, Li and Guo.
Guarda su PubMed -
Atrial fibroblast-derived exosomal miR-21 upregulate myocardial KCa3.1 via the PI3K-Akt pathway during rapid pacing.
Heliyon2024 Jul;10(13):e33059. doi: e33059.
Yuntao Fu, Jinjun Liang, Hua Fen Liu, Huiyu Chen, Dishiwen Liu, Zhen Cao, Wang Youcheng, Wang Xuewen, Ke Yuanjia, Yanni Cheng, Kexin Guo, Zhibin Peng, Mei Yang, Zhao Qingyan,
Abstract
BACKGROUND:
Fibroblast-derived exosomes can regulate the electrical remodeling of cardiomyocytes, and the intermediate-conductance calcium-activated potassium channel (KCa3.1) is important in atrial electrical remodeling. However, the underlying molecular mechanisms remain unclear. This study aimed to investigate the regulation of cardiac electrophysiology by exosomes linked to KCa3.1.
METHODS:
Atrial myocytes (AMs) and atrial fibroblasts were isolated from Sprague-Dawley suckling rats and cultured individually. The cellular atrial fibrillation (AF) model was established via electrical stimulation (1.0 v/cm, 10 Hz), and fibroblast-derived exosomes were isolated via ultracentrifugation. Exosomes were co-cultured with AMs to investigate their influences on KCa3.1 and the underlying mechanisms. Nanoparticle tracking analysis and transmission electron microscopy were used to measure exosome particle sizes and concentrations. Whole-cell patch clamp was applied to record the current density of KCa3.1 and action potential duration (APD). The expression of miR-21-5p was detected by reverse-transcription polymerase chain reaction (RT-PCR). Western blotting or immunofluorescence was used to measure the expression of exosomal markers, Akt phosphorylation, and KCa3.1.
RESULTS:
Rapid pacing promoted the secretion of exosomes from atrial fibroblasts and miR-21-5p expression in atrial fibroblasts and exosomes. KCa3.1 protein expression and current density significantly increased, and APD50 and APD90 were sharply shortened after rapid pacing in AMs. TRAM-34 (KCa3.1 blocker) extended APD and reduced susceptibility to AF. KCa3.1 and P-AKT expressions were amplified after co-culturing AMs with exosomes secreted by atrial fibroblasts. In contrast, the increase in KCa3.1 expression was reversed after the cells were co-cultured with exosomes secreted by atrial fibroblasts that were transfected with miR-21-5p inhibitors or after the use of LY294002, a PI3K/Akt pathway inhibitor.
CONCLUSIONS:
Rapid pacing promoted the secretion of exosomes from fibroblasts, and miR-21-5p was upregulated in exosomes. Moreover, the miR-21-5p-enriched exosomes upregulated KCa3.1 expression in AMs via the PI3K/Akt pathway.
© 2024 The Authors.
Guarda su PubMed -
The protective role of GATA6 pericardial macrophages in pericardial inflammation.
iScience2024 Jul;27(7):110244. doi: 110244.
Hughes David M, Won Taejoon, Talor Monica V, Kalinoski Hannah M, Jur?ová Ivana, Szárszoi Ondrej, St?í? Ilja, ?urnová Lenka, Bracamonte-Baran William, Melenovský Vojt?ch, ?iháková Daniela,
Abstract
Prior research has suggested that GATA6 pericardial macrophages may traffic to the myocardium to prevent interstitial fibrosis after myocardial infarction (MI), while subsequent literature claims that they do not. We demonstrate that GATA6 pericardial macrophages are critical for preventing IL-33 induced pericarditis and attenuate trafficking of inflammatory monocytes and granulocytes to the pericardial cavity after MI. However, absence of GATA6 macrophages did not affect myocardial inflammation due to MI or coxsackievirus-B3 induced myocarditis, or late-stage cardiac fibrosis and cardiac function post MI. GATA6 macrophages are significantly less transcriptionally active following stimulation compared to bone marrow-derived macrophages and do not induce upregulation of inflammatory markers in fibroblasts. This suggests that GATA6 pericardial macrophages attenuate inflammation through their interactions with surrounding cells. We therefore conclude that GATA6 pericardial macrophages are critical in modulating pericardial inflammation, but do not play a significant role in controlling myocardial inflammation or fibrosis.
© 2024 The Author(s).
Guarda su PubMed -
Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases.
Nat Biotechnol2024 Jul;():. doi: 10.1038/s41587-024-02324-x.
Ferreira da Silva Joana, Tou Connor J, King Emily M, Eller Madeline L, Rufino-Ramos David, Ma Linyuan, Cromwell Christopher R, Metovic Jasna, Benning Friederike M C, Chao Luke H, Eichler Florian S, Kleinstiver Benjamin P,
Abstract
Genome editing technologies based on DNA-dependent polymerases (DDPs) could offer several benefits compared with other types of editors to install diverse edits. Here, we develop click editing, a genome writing platform that couples the advantageous properties of DDPs with RNA-programmable nickases to permit the installation of a range of edits, including substitutions, insertions and deletions. Click editors (CEs) leverage the 'click'-like bioconjugation ability of HUH endonucleases with single-stranded DNA substrates to covalently tether 'click DNA' (clkDNA) templates encoding user-specifiable edits at targeted genomic loci. Through iterative optimization of the modular components of CEs and their clkDNAs, we demonstrate the ability to install precise genome edits with minimal indels in diverse immortalized human cell types and primary fibroblasts with precise editing efficiencies of up to ~30%. Editing efficiency can be improved by rapidly screening clkDNA oligonucleotides with various modifications, including repair-evading substitutions. Click editing is a precise and versatile genome editing approach for diverse biological applications.
© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.
Guarda su PubMed -
Jiedu Tongluo Decoction Attenuates Myocardial Fibrosis through Inhibition of the TGF-?1/Smad2/3 Pathway.
Altern Ther Health Med2024 Jul;():. doi: AT9589.
Hui Ju, Li Le, Zhang Huan, Wang Bin, Shi Jiaxin, Song Jing, Guo Jiajuan, Li Xiangjun,
Abstract
Jiedu Tongluo (JDTL) Decoction is a traditional Chinese medicine formula containing three herbal ingredients. It is widely used to treat myocardial fibrosis (MF). This study aimed to investigate the molecular mechanism of JDTL Decoction's effect on MF. In this study, 6 compounds of JDTL Decoction were identified by HPLC. HE and Masson staining showed that in the isoproterenol hydrochloride-induced MF rat model, JDTL treatment can protect the myocardial structure and inhibit the expression of collagen III. The immunohistochemistry results also showed that JDTL treatment can significantly reduce vimentin and ?-SMA expression, TGF-?1 expression, and phosphorylation of Smad2/3 in the rat MF model. RCF, a rat cardiac fibroblast cell line, was used as a tool for in vitro study. Using the methods of hydroxyproline detection, MTT, wound healing test, western blot, and double immunofluorescence staining, our in vitro study confirmed the inhibitory effects of JDTL Decoction on proliferation, migration, and trans-difference ability of RCF cells, as well as the molecular mechanisms underlying the inhibitory effects of JDTL Decoction, including the inhibition of TGF-?1/Smad2/3 pathway through down-regulation of TGF-?1 expression and phosphorylation of Smad2/3 as well as the inhibition of the expression of vimentin and ?-SMA. In conclusion, JDTL Decoction can prolong the process of myocardial fibrosis through the inhibition of the TGF?1/Smad2/3 signaling pathway.
Guarda su PubMed -
Generation and Characterization of hiPSC-Derived Vascularized-, Perfusable Cardiac Microtissues-on-Chip.
Curr Protoc2024 Jul;4(7):e1097. doi: 10.1002/cpz1.1097.
Arslan Ulgu, van den Hil Francijna E, Mummery Christine L, Orlova Valeria,
Abstract
In the heart in vivo, vasculature forms a semi-permeable endothelial barrier for selective nutrient and (immune) cell delivery to the myocardium and removal of waste products. Crosstalk between the vasculature and the heart cells regulates homeostasis in health and disease. To model heart development and disease in vitro it is important that essential features of this crosstalk are captured. Cardiac organoid and microtissue models often integrate endothelial cells (ECs) to form microvascular networks inside the 3D structure. However, in static culture without perfusion, these networks may fail to show essential functionality. Here, we describe a protocol to generate an in vitro model of human induced pluripotent stem cell (hiPSC)-derived vascularized cardiac microtissues on a microfluidic organ-on-chip platform (VMToC) in which the blood vessels are perfusable. First, prevascularized cardiac microtissues (MT) are formed by combining hiPSC-derived cardiomyocytes, ECs, and cardiac fibroblasts in a pre-defined ratio. Next, these prevascularized MTs are integrated in the chips in a fibrin hydrogel containing additional vascular cells, which self-organize into tubular structures. The MTs become vascularized through anastomosis between the pre-existing microvasculature in the MT and the external vascular network. The VMToCs are then ready for downstream structural and functional assays and basic characterization. Using this protocol, cardiac MTs can be efficiently and robustly vascularized and perfused within 7 days. In vitro vascularized organoid and MT models have the potential to transition current 3D cardiac models to more physiologically relevant organ models that allow the role of the endothelial barrier in drug and inflammatory response to be investigated. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Generation of VMToC Support Protocol 1: Functional Characterization of VMToC Support Protocol 2: Structural Characterization of VMToC.
© 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC.
Guarda su PubMed -
Pathological insights into activin A: Molecular underpinnings and therapeutic prospects in various diseases.
Int Immunopharmacol2024 Jul;139():112709. doi: 10.1016/j.intimp.2024.112709.
Kundra Sejal, Kaur Rupinder, Pasricha Chirag, Kumari Pratima, Gurjeet Singh Thakur, Singh Ravinder,
Abstract
Activin A (Act A) is a member of the TGF? (transforming growth factor ?) superfamily. It communicates via the Suppressor of Mothers against Decapentaplegic Homolog (SMAD2/3) proteins which govern processes such as cell proliferation, wound healing, apoptosis, and metabolism. Act A produces its action by attaching to activin receptor type IIA (ActRIIA) or activin receptor type IIB (ActRIIB). Increasing circulating Act A increases ActRII signalling, which on phosphorylation initiates the ALK4 (activin receptor-like kinase 4) type 1 receptor which further turns on the SMAD pathway and hinders cell functioning. Once triggered, this route leads to gene transcription, differentiation, apoptosis, and extracellular matrix (ECM) formation. Act A also governs the immunological and inflammatory responses of the body, as well as cell death. Moreover, Act A levels have been observed to elevate in several disorders like renal fibrosis, CKD, asthma, NAFLD, cardiovascular diseases, cancer, inflammatory conditions etc. Here, we provide an update on the recent studies relevant to the role of Act A in the modulation of various pathological disorders, giving an overview of the biology of Act A and its signalling pathways, and discuss the possibility of incorporating activin-A targeting as a novel therapeutic approach for the control of various disorders. Pathways such as SMAD signaling, in which SMAD moves to the nucleus by making a complex and leads to tissue fibrosis in CKD, STAT3, which drives renal fibroblast activity and the production of ECM, Kidney injury molecule (KIM-1) in the synthesis, deposition of ECM proteins, SERCA2a (sarcoplasmic reticulum Ca ATPase) in cardiac dysfunction, and NF-?B (Nuclear factor kappa-light-chain-enhancer of activated B cells) in inflammation are involved in Act A signaling, have also been discussed.
Copyright © 2024 Elsevier B.V. All rights reserved.
Guarda su PubMed -
IL-17 is associated with disease severity and targetable inflammatory processes in heart failure.
ESC Heart Fail2024 Jul;():. doi: 10.1002/ehf2.14968.
Baumhove Lukas, van Essen Bart J, Dokter Martin M, Zijlstra Sietske N, Deiman Frederik E, Laman Jon D, Lang Chim C, Verstappen Gwenny M P J, van Veldhuisen Dirk J, van der Meer Peter, Bomer Nils, Voors Adriaan A,
Abstract
AIMS:
Heart failure (HF) is recognized as an inflammatory disease in which cytokines play an important role. In animal HF models, interleukin-17A (IL-17) has been linked to deterioration of cardiac function and fibrosis, whereas knock-out of IL-17 showed beneficial cardiac effects. However, there is limited evidence of IL-17 involvement in patients with HF. This study aims to investigate the clinical characteristics, outcomes, and pathophysiological processes associated with circulating IL-17 concentrations in patients with HF.
METHODS AND RESULTS:
IL-17 was measured by ELISA in 2082 patients diagnosed with HF along with 363 circulating proteins using proximity extension assay technology for differential expression and pathway analysis. Data were validated in an independent cohort of 1737 patients with HF. Patients with elevated IL-17 concentrations had more severe HF, as reflected by more frequent current or previous hospitalizations for HF, higher New York Heart Association functional class (NYHA) and higher levels of N-terminal pro-B-type natriuretic peptide (NT-proBNP). High IL-17 concentrations were independently associated with an increased risk of hospitalization for HF and mortality. In both cohorts, the most strongly up-regulated proteins in patients with high IL-17 were fibroblast growth factor 21 (FGF-21), interleukin-6 (IL-6), C-X-C motif chemokine ligand 13 (CXCL13), tumour necrosis factor receptor superfamily member 6B (TNFRSF6B) and interleukin-1 receptor antagonist (IL-1RA). Pathway over-representation analysis showed increased activity of pathways related to lymphocyte-mediated immunity, leukocyte activation and regulation of the immune response.
CONCLUSIONS:
In patients with HF, elevated IL-17 concentrations indicate more severe HF and increased activity of inflammatory processes known to be involved in the pathophysiology of HF. IL-17 might hold potential for identifying and targeting inflammation in HF.
© 2024 The Author(s). ESC Heart Failure published by John Wiley & Sons Ltd on behalf of European Society of Cardiology.
Guarda su PubMed -
Histological evaluation of cardiac remodelling in equine athletes.
Sci Rep2024 Jul;14(1):16709. doi: 16709.
Nath L C, Saljic A, Buhl R, Elliott A, La Gerche A, Ye C, Schmidt Royal H, Lundgren Virklund K, Agbaedeng T A, Stent A, Franklin S,
Abstract
Approximately 1-2 per 100,000 young athletes die from sudden cardiac death (SCD) and extreme exercise may be associated with myocardial scar and arrhythmias. Racehorses have a high prevalence of atrial fibrillation (AF) and SCD but the presence of myocardial scar and inflammation has not been evaluated. Cardiac tissues from the left (LAA) and right (RAA) atrial appendages, left ventricular anterior (LVAPM) and posterior (LVPPM) papillary muscles, and right side of the interventricular septum (IVS-R) were harvested from racehorses with sudden cardiac death (SCD, n?=?16) or other fatal injuries (OFI, n?=?17), constituting the athletic group (ATH, n?=?33), and compared to sedentary horses (SED, n?=?10). Horses in the ATH group had myocyte hypertrophy at all sites; increased fibrosis at all sites other than the LAA; increased fibroblast infiltration but a reduction in the overall extracellular matrix (ECM) volume in the RAA, LVAPM, and IVS-R compared to SED horses. In this horse model, athletic conditioning was associated with myocyte hypertrophy and a reduction in ECM. There was an excess of fibrocyte infiltration and focal fibrosis that was not present in non-athletic horses, raising the possibility of an exercise-induced pro-fibrotic substrate.
© 2024. The Author(s).
Guarda su PubMed -
Antibacterial, mosquito larvicidal, and cytotoxicity potential of AgNPs synthesized using Pittosporum undulatum under in vitro conditions.
Environ Res2024 Jul;():119585. doi: 10.1016/j.envres.2024.119585.
Narayanan Mathiyazhagan, Alshiekheid Maha A, Saravanan Mythili,
Abstract
In this study, the phytochemical profile and silver nanoparticle (AgNP)-synthesizing ability of Pittosporum undulatum methanol extract were investigated. Furthermore, biological applications of the AgNPs, such as antibacterial effect (against Klebsiella pneumoniae, Staphylococcus aureus, Bacillus subtilis, and Escherichia coli), mosquito larvicidal effect (against Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti), and cytotoxicity (against fibroblast cell line L929) were evaluated using in vitro experiments. The phytochemical analysis revealed that the methanol extract contained cardiac glycosides, terpenoids, saponins, alkaloids, flavonoids, glycosides, coumarins, phenolics, and tannins. Furthermore, standard characterization techniques such as UV-Vis spectrometry, SEM, TEM, FTIR, and XRD confirmed that the methanol extract of P. undulatum effectively synthesized the AgNPs. The synthesized AgNPs had a spherical shape and size of 20-200 nm. The bactericidal analysis revealed that the AgNPs have dose-dependent antibacterial activity. The MTT assay showed that the AgNPs were bio-compatible up to a dosage of 250 ?g·mL in the normal fibroblast cell line L929. Furthermore, the LC values for AgNPs against larvae of An. stephensi, Cx. quinquefasciatus, and Ae. aegypti were 0.4, 4.7, and 1.2 ppm, respectively. Field trials demonstrated that the larvicidal effect was enhanced within 24-72 h, and the rate of reduction increased over time. Thus, our findings provide an ideal sustainable AgNP bio-pesticide to combat filarial, dengue, and malaria vectors.
Copyright © 2024. Published by Elsevier Inc.
Guarda su PubMed -
The roles of orphan nuclear receptor 4 group A1 and A2 in fibrosis.
Int Immunopharmacol2024 Jul;139():112705. doi: 10.1016/j.intimp.2024.112705.
Gao Lanjun, Wang Hongshuang, Fang Fang, Liu Jiazhi, Zhao Chenchen, Niu Jieqi, Wang Zheng, Zhong Yan, Wang Xiangting,
Abstract
Fibrosis is not a disease but rather an outcome of the pathological tissue repair response. Many myofibroblasts are activated which lead to the excessive accumulation of extracellular matrix components such as collagen and fibronectin with fibrosis. A variety of organs, including kidney, liver, lung, heart and skin, can undergo fibrosis under the stimulation of exogenous or endogenous pathogenic factors. The orphan nuclear receptor 4 group A1 (NR4A1) and nuclear receptor 4 group A2?NR4A2?are belong to the nuclear receptor subfamily and inhibit the occurrence and development of fibrosis. NR4A1 is an inhibitory factor of TGF-? signaling transduction. Overexpression of NR4A1 in fibroblasts can reduce TGF-? induced collagen deposition and fibrosis related gene expression. Here, we summarize the current research progress on the NR4A1/2 and fibrosis, providing reference for the treatment of fibrosis.
Copyright © 2024 Elsevier B.V. All rights reserved.
Guarda su PubMed -
Polycystin-1 loss of function increases susceptibility to atrial fibrillation through impaired DNA damage response.
bioRxiv2024 Jul;():. doi: 2024.07.08.602618.
Hendrickson Troy, Abigail Giese Abigail, Fiedler Matthew, Perez William, Reyes-Sanchez Ernesto, Reyes-Lozano Monserrat, Wang Sufen, Venegas-Zamora Leslye, Provasek Vincent, El-Essawi Aschraf, Breitenbach Ingo, Fakuade Funsho E, Kutschka Ingo, Schiattarella Gabriele G, Voigt Niels, Valderrabano Miguel, Altamirano Francisco,
Abstract
BACKGROUND:
The increasing prevalence of atrial fibrillation (AF) and chronic kidney diseases highlights the need for a deeper comprehension of the molecular mechanisms linking them. Mutations in PKD1, the gene encoding Polycystin-1 (PKD1 or PC1), account for 85% of autosomal dominant polycystic kidney disease (ADPKD) cases. This disease often includes cardiac complications such as AF. In cardiomyocytes, PC1 deletion reduces hypertrophic response to pressure overload but promotes baseline ventricular dysfunction, while deletion in fibroblasts ameliorates post-myocardial infarction fibrosis. Despite its known cardiac impact, the role of PC1 in atrial cardiomyocytes and arrhythmias is less understood. Here, we sought to investigate the role of PC1 in AF.
METHODS:
We used intracardiac programmed stimulation and optical mapping to evaluate AF inducibility in two mouse models, Pkd1 R3277C, which recapitulates human ADPKD progression, and cardiomyocyte-specific Pkd1 deletion, and their respective controls. Isolated adult mouse atrial cardiomyocytes, human iPSC-derived atrial cardiomyocytes (hiPSC-aCM), and HL-1 cells served as in vitro cellular models. Molecular mechanisms were evaluated using optical mapping and molecular and biochemical approaches.
RESULTS:
Loss-of-function PC1 mutations significantly increased AF susceptibility in vivo and facilitated local reentry in ex vivo left atrial appendages. Comprehensive in vitro experiments supported a direct effect of PC1 in atrial cardiomyocytes. PC1-deficient monolayers exhibited increased arrhythmic events, escalating into reentrant spiral waves post-tachypacing. Transcriptomics analysis revealed PC1-dependent regulation of DNA repair, with PC1 deficiency leading to increased DNA damage under stress. PARP1 inhibitors or nicotinamide riboside, which counteract DNA damage-related metabolic consequences, reduced in vitro arrhythmias PC1-deficient monolayers. Overexpression of the C-terminus of PC1 had the opposite effects in DNA repair genes, suggesting its regulatory effects in atrial cardiomyocytes through retinoblastoma/E2F. Analyses of human atrial tissue from non-ADPKD patients showed reduced levels of mature PC1, suggesting a broader relevance of impaired PC1 in AF.
CONCLUSIONS:
Impaired PC1 increases in vivo AF inducibility under programmed electrical stimulation and promotes in vitro arrhythmias in hiPSC-aCM and HL-1 cells. Our findings indicate that PC1 protects against DNA damage to reduce AF susceptibility.
Guarda su PubMed
![](/img/pubmed.png)