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1.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 35(4): 376-380, 2019 Jul 28.
Artigo em Chinês | MEDLINE | ID: mdl-31701727

RESUMO

OBJECTIVE: To investigate whether salidroside (Sal) plays a part in protecting myocardial cell through reducing the myocardial ischemia and the apoptosis pathway of both death receptors and mitochondria in acute exhausted rats. METHODS: Male SD rats were randomly divided into 4 groups (n=6): control group(Con), acute exhaustive swimming group (EE), low-dose and high-dose Sal pre-treatment exhaustive swimming group (SLE, SHE). Rats were treated with Sal solution (15 or 30 mg/(kg·d)) or 0.9%NaCl (3 ml/(kg·d)) by intraperitoneal injection for 15 d, respectively. The Con group did not carry out swimming training. The next day after the end of intraperitoneal administration, the rats in EE, SLE and SHE group were forced to swim until they were exhausted followed the standard of Thomas. After the end of exhaustive exercise, the rats were anesthetized and the blood samples and hearts were collected immediately. The myocardial ischemia and hypoxia area and myocardial apoptosis index (AI) were also observed. Serum ischemia modified albumin (IMA), cardiac troponin I (cTnI), brain natriuretic peptide(BNP) and myocardial cell Bcl-2-associated X protein (Bax), B-cell lymphoma-2 (Bcl-2) were determined. The expressions of myocardial TNF receptor superfamily member 6 (Fas), cytochrome C (Cyto-c), aspartate proteolytic enzyme-3(Caspase-3), aspartate proteolytic enzyme-8(Caspase-8), and aspartate proteolytic enzyme-9(Caspase-9) were detected. RESULTS: Compared with the Con group, the myocardial ischemia and hypoxia area in EE group was increased significantly. The serum levels of IMA, cTnI and BNP, AI and Bax levels and cardiac Fas, Cyto-C, Caspase-3, Caspase-8 and Caspase-9 protein expressions of EE group were also increased significantly (P<0.01), while the protein expression of Bcl-2 in cardiac tissues was decreased significantly (P<0.01). Compared with the EE group, the myocardial ischemia and hypoxia area, serum levels of IMA, cTnI and BNP, AI and Bax levels, and the protein expressions of cardiac Fas, Cyto-C, Caspase-3, Caspase-8 and Caspase-9 in Sal group were all decreased significantly(P<0.01). while the protein expression of cardiac Bcl-2 in Sal group were increased significantly (P<0.01). CONCLUSION: Sal plays a role in protecting myocardial cell through reducing the myocardial ischemia and inhibiting myocardial cell apoptosis in exhaustive exercise rats. The mechanism of reducing myocardial cell apoptosis may be related to inhibiting the expressions of Fas, Cyto-C, Caspase-3, Caspase-8, Caspase-9 and increasing the expression of Bcl-2.


Assuntos
Apoptose , Fadiga/fisiopatologia , Glucosídeos/farmacologia , Coração/efeitos dos fármacos , Isquemia Miocárdica/tratamento farmacológico , Fenóis/farmacologia , Animais , Biomarcadores/sangue , Feminino , Masculino , Miocárdio/citologia , Condicionamento Físico Animal , Ratos , Ratos Sprague-Dawley
2.
Int J Sports Med ; 40(14): 921-930, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31614382

RESUMO

Myocardial damage due to dysfunctional myocardium has been increasing, and the prognosis of pharmacological and device-based therapies remain poor. Isl1-expressing cells were thought to be progenitor cells for cardiomyocyte proliferation after specific stimuli. However, the true origin of the proliferating myocardiac cells and the role of Isl1 in adult mammals remain unresolved. In this study, Isl1-CreERT2 knock-in mouse model was constructed using CRISPR/Cas9 technology. Using tamoxifen-inducible Isl1-CreERT/Rosa26R-LacZ system, Isl1+cells and their progeny were permanently marked by lacZ-expression. X-gal staining, immunostaining, and quantitative PCR were then used to reveal the fate of Isl1+cells under physiological and exercise conditions in mouse hearts from embryonic stage to adulthood. Isl1+cells were found to localize to the sinoatrial node, atrioventricular node, cardiac ganglia, aortic arch, and pulmonary roots in adult mice heart. However, they did not act as cardiac progenitor cells under physiological and exercise conditions. Although Isl1+cells showed progenitor cell properties in early mouse embryos (E7.5), this ability was lost by E9.5. Furthermore, although the proliferation and regeneration of heart cell was observed in response to exercise, the cells associated were not Isl1 positive.


Assuntos
Coração/fisiologia , Proteínas com Homeodomínio LIM/genética , Miocárdio/citologia , Miócitos Cardíacos/citologia , Condicionamento Físico Animal , Células-Tronco/fisiologia , Fatores de Transcrição/genética , Animais , Proteína 9 Associada à CRISPR , Sistemas CRISPR-Cas , Proliferação de Células/fisiologia , Mapeamento Cromossômico , Coração/embriologia , Coração/crescimento & desenvolvimento , Masculino , Camundongos Endogâmicos C57BL
3.
Mater Sci Eng C Mater Biol Appl ; 104: 109921, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31500009

RESUMO

Graphene based nanomaterials are promising candidates for cardiac tissue engineering due to the excellent electrical and mechanical properties and the robust surface chemistry. This research was designed to investigate the physicochemical and biological effects of increasing concentration of reduced graphene oxide (rGO) coating on collagen (Col) scaffolds as well as their antibacterial properties. Enhanced GO coating content to 400 µg/ml and its reduction showed improvement of HUVECs viability, however, following reduction of more GO concentration, decreased cell viability was observed. Compared with the Col counterpart, electroactive containing rGO scaffolds upregulated cardiac gene expression involve in electrical coupling (Cx43), muscle contraction and relaxation (troponin-T) and cytoskeleton alignment (actinin-4) after 7 days even without external electrical stimulation. rGO coating significantly improved mechanical properties and the electroactivity of the Col scaffolds reaching to 1100 ±â€¯31 kPa and 4 × 10-4 ±â€¯1.20 S/m for GO concentration of 800 µg/ml, respectively. Also, the antibacterial properties of Col-rGO-400 scaffolds against Escherichia coli, Staphylococcus aureus and Streptococcus pyogenes were confirmed by culture and FESEM observation. Taken together, the results indicated that rGO coating presents promising properties to Col scaffolds providing a desirable micro environment for cardiomyocytes coupling and gene upregulation as well as antibacterial activities for cardiac patch application.


Assuntos
Antibacterianos/farmacologia , Eletricidade , Grafite/farmacologia , Miocárdio/citologia , Tecidos Suporte/química , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Aderência Bacteriana/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/citologia , Humanos , Testes de Sensibilidade Microbiana , Oxirredução , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
4.
Adv Exp Med Biol ; 1169: 141-178, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31487023

RESUMO

Cardiac biology and heart regeneration have been intensively investigated and debated in the last 15 years. Nowadays, the well-established and old dogma that the adult heart lacks of any myocyte-regenerative capacity has been firmly overturned by the evidence of cardiomyocyte renewal throughout the mammalian life as part of normal organ cell homeostasis, which is increased in response to injury. Concurrently, reproducible evidences from independent laboratories have convincingly shown that the adult heart possesses a pool of multipotent cardiac stem/progenitor cells (CSCs or CPCs) capable of sustaining cardiomyocyte and vascular tissue refreshment after injury. CSC transplantation in animal models displays an effective regenerative potential and may be helpful to treat chronic heart failure (CHF), obviating at the poor/modest results using non-cardiac cells in clinical trials. Nevertheless, the degree/significance of cardiomyocyte turnover in the adult heart, which is insufficient to regenerate extensive damage from ischemic and non-ischemic origin, remains strongly disputed. Concurrently, different methodologies used to detect CSCs in situ have created the paradox of the adult heart harboring more than seven different cardiac progenitor populations. The latter was likely secondary to the intrinsic heterogeneity of any regenerative cell agent in an adult tissue but also to the confusion created by the heterogeneity of the cell population identified by a single cell marker used to detect the CSCs in situ. On the other hand, some recent studies using genetic fate mapping strategies claimed that CSCs are an irrelevant endogenous source of new cardiomyocytes in the adult. On the basis of these contradictory findings, here we critically reviewed the available data on adult CSC biology and their role in myocardial cell homeostasis and repair.


Assuntos
Células-Tronco Adultas , Miocárdio , Animais , Diferenciação Celular , Miocárdio/citologia , Miócitos Cardíacos/citologia
5.
Nat Protoc ; 14(10): 2781-2817, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31492957

RESUMO

The application of tissue-engineering approaches to human induced pluripotent stem (hiPS) cells enables the development of physiologically relevant human tissue models for in vitro studies of development, regeneration, and disease. However, the immature phenotype of hiPS-derived cardiomyocytes (hiPS-CMs) limits their utility. We have developed a protocol to generate engineered cardiac tissues from hiPS cells and electromechanically mature them toward an adult-like phenotype. This protocol also provides optimized methods for analyzing these tissues' functionality, ultrastructure, and cellular properties. The approach relies on biological adaptation of cultured tissues subjected to biomimetic cues, applied at an increasing intensity, to drive accelerated maturation. hiPS cells are differentiated into cardiomyocytes and used immediately after the first contractions are observed, when they still have developmental plasticity. This starting cell population is combined with human dermal fibroblasts, encapsulated in a fibrin hydrogel and allowed to compact under passive tension in a custom-designed bioreactor. After 7 d of tissue formation, the engineered tissues are matured for an additional 21 d by increasingly intense electromechanical stimulation. Tissue properties can be evaluated by measuring contractile function, responsiveness to electrical stimuli, ultrastructure properties (sarcomere length, mitochondrial density, networks of transverse tubules), force-frequency and force-length relationships, calcium handling, and responses to ß-adrenergic agonists. Cell properties can be evaluated by monitoring gene/protein expression, oxidative metabolism, and electrophysiology. The protocol takes 4 weeks and requires experience in advanced cell culture and machining methods for bioreactor fabrication. We anticipate that this protocol will improve modeling of cardiac diseases and testing of drugs.


Assuntos
Células-Tronco Pluripotentes Induzidas/citologia , Miocárdio , Engenharia Tecidual/métodos , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Coração/fisiologia , Humanos , Miocárdio/citologia , Miócitos Cardíacos/citologia , Miócitos Cardíacos/fisiologia
6.
Pediatr Cardiol ; 40(7): 1410-1418, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31399780

RESUMO

Numerous RNA-binding proteins (RBPs) are expressed in the heart, and mutations in several RBPs have been implicated in cardiovascular disease through genetic associations, animal modeling, and mechanistic studies. However, the functions of many more cardiac RBPs, and their relevance to disease states, remain to be elucidated. Recently, we have initiated studies to characterize the functions of the RBPs RBPMS and RBPMS2 in regulating myocardial biology in zebrafish and higher vertebrate species. These studies began when we learned, using an unbiased gene discovery approach, that rbpms2a and rbpms2b in zebrafish are robust markers of embryonic myocardium. This observation, which is consistent with published data, suggests that the encoded proteins are likely to be performing critical functions in regulating one or more aspects of cardiomyocyte differentiation, proliferation, survival, and/or contractility. This notion is supported by recent reports demonstrating that zebrafish embryos with disrupted Rbpms2 function exhibit gross signs of cardiac distress. Interestingly, a 20-year-old study determined that myocardial tissue from the frog, chick, and mouse also express high levels of Rbpms and/or Rbpms2, which is suggestive of evolutionary conservation of function. In this review, we will provide a historical account of how RBPMS and RBPMS2 genes were discovered, attempt to clarify some potentially confusing nomenclature, and summarize published observations that inform our ongoing studies.


Assuntos
Miocárdio/citologia , Proteínas de Ligação a RNA/metabolismo , Animais , Diferenciação Celular , Humanos , Camundongos , Proteínas de Ligação a RNA/genética , Peixe-Zebra
7.
BMC Mol Cell Biol ; 20(1): 32, 2019 08 13.
Artigo em Inglês | MEDLINE | ID: mdl-31409295

RESUMO

BACKGROUND: Endothelial cells (ECs) play a key role in tissue homeostasis, in several pathological conditions, and specifically in the control of vascular functions. ECs are frequently used as in vitro model systems for cardiovascular studies and vascular biology. The porcine model is commonly used in human clinical cardiovascular studies. Currently, however, there is no robust protocol for the isolation of porcine heart ECs. We have developed a fast isolation protocol, which is cost effective, takes only 1-2 h, and produces EC purity of over 97%. This protocol is optimized for porcine hearts but can be adapted for use with other large animals. METHODS: Heart is washed by flushing with PBS, whereafter endothelial cells are detached by collagenase incubation and the cells can then be collected immediately after the incubation and plated within an hour after the heart is isolated from a pig. RESULTS: The swiftness of the protocol limits changes in the phenotype and RNA expression profile of the cells. Cells were identified as ECs with CD31 (PECAM-1) antibody immunostaining. Functionality of ECs were ensured with in vitro angiogenesis assay. The purity of the ECs was verified by using fluorescence assisted cell sorting (FACS) with the CD31 antibody. CONCLUSION: We developed a new, fast, and cost-effective isolation method for pig heart ECs. Successful isolation of pure ECs is a prerequisite for several cardiovascular and vascular biology studies.


Assuntos
Doenças Cardiovasculares/patologia , Biologia Celular , Separação Celular/métodos , Células Endoteliais/citologia , Genômica , Miocárdio/citologia , Transcriptoma/genética , Animais , Células Cultivadas , Feminino , Suínos
8.
Bull Exp Biol Med ; 167(3): 315-319, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31346863

RESUMO

Vitronectin, extracellular matrix protein, plays an important role in embryonic development and in organ and tissue reparation. A unique characteristic of vitronectin is specific binding of various biological molecules, including urokinase receptor (uPAR), extracellular matrix components, adhesion receptors, growth factors, thus supporting the modulation of cell behavior. Vitronectin is in fact not found in intact myocardium, while after infarction its level increases significantly, which correlates with accumulation of uPAR+ progenitor cardiac cells in the focus. The cells isolated from the heart of wild type mice are characterized by higher adhesion to vitronectin than progenitor cardiac cells from the myocardium of uPAR knockout mice. In addition, inhibition of urokinase receptor with specific antibodies on the surface of the progenitor cardiac cells of wild type mice leads to attenuation of their adhesive activity and flattening on vitronectin matrix, which can be important for their distribution in the postinfarction myocardium and realization of the reparative functions.


Assuntos
Adesão Celular/fisiologia , Miocárdio/metabolismo , Receptores de Ativador de Plasminogênio Tipo Uroquinase/fisiologia , Células-Tronco/fisiologia , Vitronectina/metabolismo , Animais , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Infarto do Miocárdio/patologia , Miocárdio/citologia , Receptores de Ativador de Plasminogênio Tipo Uroquinase/antagonistas & inibidores , Receptores de Ativador de Plasminogênio Tipo Uroquinase/genética
9.
Mater Sci Eng C Mater Biol Appl ; 103: 109712, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31349433

RESUMO

Electrospun fibers based on combinations of poly(ε-caprolactone) (PCL) and poly(glycerol sebacate) (PGS) have been studied for applications in cardiac tissue engineering. The aim of the present study is to demonstrate the fabrication of PCL and PGS prepolymer or mildly crosslinked PGS by electrospinning using less toxic solvents, like acetic acid, as opposed to conventional solvents such as chloroform or dichloromethane. The morphological and physiochemical properties and degradation susceptibility of the fiber mats were determined. A cell study using ST2 cells was performed to assess cytocompatibility. Neat PCL and PCL/PGS blends showed defect-free microstructures, whereby the average fiber diameter increased with the addition of PGS (0.8 ±â€¯0.3 µm and 1.3 ±â€¯0.7 µm, respectively). The mechanical properties were tested at 23 °C and 37 °C and showed higher values compared to native human myocardium. Degradation studies revealed a fast PGS degradation but the mildly cross-linked PGS resulted in acidification of the degradation medium. The biocompatibility was significantly increased when the samples were disinfected in 70% v/v ethanol in comparison to ultra-violet light disinfection. PCL/PGS fibers fabricated with acetic acid proved to be potentially suitable for application in cardiac tissue engineering based on their mechanical properties and biodegradability.


Assuntos
Decanoatos/química , Glicerol/análogos & derivados , Teste de Materiais , Miocárdio/metabolismo , Poliésteres/química , Polímeros/química , Engenharia Tecidual , Tecidos Suporte/química , Linhagem Celular , Glicerol/química , Humanos , Miocárdio/citologia
10.
Nat Commun ; 10(1): 3027, 2019 07 09.
Artigo em Inglês | MEDLINE | ID: mdl-31289275

RESUMO

Fibrosis accompanying wound healing can drive the failure of many different organs. Activated fibroblasts are the principal determinants of post-injury pathological fibrosis along with physiological repair, making them a difficult therapeutic target. Although activated fibroblasts are phenotypically heterogeneous, they are not recognized as distinct functional entities. Using mice that express GFP under the FSP1 or αSMA promoter, we characterized two non-overlapping fibroblast subtypes from mouse hearts after myocardial infarction. Here, we report the identification of FSP1-GFP+ cells as a non-pericyte, non-hematopoietic fibroblast subpopulation with a predominant pro-angiogenic role, characterized by in vitro phenotypic/cellular/ultrastructural studies and in vivo granulation tissue formation assays combined with transcriptomics and proteomics. This work identifies a fibroblast subtype that is functionally distinct from the pro-fibrotic αSMA-expressing myofibroblast subtype. Our study has the potential to shift our focus towards viewing fibroblasts as molecularly and functionally heterogeneous and provides a paradigm to approach treatment for organ fibrosis.


Assuntos
Fibroblastos/metabolismo , Infarto do Miocárdio/patologia , Miocárdio/patologia , Neovascularização Fisiológica/fisiologia , Cicatrização/fisiologia , Actinas/genética , Actinas/metabolismo , Animais , Transplante de Medula Óssea , Proteínas de Ligação ao Cálcio/metabolismo , Diferenciação Celular , Modelos Animais de Doenças , Fibroblastos/patologia , Fibrose/etiologia , Fibrose/patologia , Proteínas de Fluorescência Verde/genética , Células Endoteliais da Veia Umbilical Humana , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Infarto do Miocárdio/etiologia , Infarto do Miocárdio/fisiopatologia , Miocárdio/citologia , Proteína A4 de Ligação a Cálcio da Família S100/genética , Proteína A4 de Ligação a Cálcio da Família S100/metabolismo , Quimeras de Transplante
11.
Cells ; 8(7)2019 07 04.
Artigo em Inglês | MEDLINE | ID: mdl-31277520

RESUMO

Coronary artery disease is the most common form of cardiovascular diseases, resulting in the loss of cardiomyocytes (CM) at the site of ischemic injury. To compensate for the loss of CMs, cardiac fibroblasts quickly respond to injury and initiate cardiac remodeling in an injured heart. In the remodeling process, cardiac fibroblasts proliferate and differentiate into myofibroblasts, which secrete extracellular matrix to support the intact structure of the heart, and eventually differentiate into matrifibrocytes to form chronic scar tissue. Discovery of direct cardiac reprogramming offers a promising therapeutic strategy to prevent/attenuate this pathologic remodeling and replace the cardiac fibrotic scar with myocardium in situ. Since the first discovery in 2010, many progresses have been made to improve the efficiency and efficacy of reprogramming by understanding the mechanisms and signaling pathways that are activated during direct cardiac reprogramming. Here, we overview the development and recent progresses of direct cardiac reprogramming and discuss future directions in order to translate this promising technology into an effective therapeutic paradigm to reverse cardiac pathological remodeling in an injured heart.


Assuntos
Reprogramação Celular/genética , Doença da Artéria Coronariana/terapia , Miocárdio/patologia , Medicina Regenerativa/métodos , Fatores de Transcrição/genética , Animais , Doença da Artéria Coronariana/patologia , Modelos Animais de Doenças , Matriz Extracelular/patologia , Fibrose , Regulação da Expressão Gênica , Vetores Genéticos/administração & dosagem , Vetores Genéticos/genética , Humanos , Injeções Intralesionais , Miocárdio/citologia , Miócitos Cardíacos/fisiologia , Miofibroblastos/fisiologia , Transdução de Sinais/genética , Fatores de Transcrição/metabolismo
12.
Mol Med Rep ; 20(3): 2403-2409, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31257537

RESUMO

The aim of the present study was to explore the feasibility of the construction of engineered myocardial tissues in vitro with cardiomyocyte­like cells derived from bone marrow mesenchymal stem cells (BMMSCs) and a polylactic­co­glycolic acid (PLGA) polymer. The PLGA polymer was sheared into square pieces (10x10x1 mm), sterilized by Co60 irradiation, and hydrated in Dulbecco's modified Eagle's medium for 1 h. BMMSCs were isolated from the bone marrow of Sprague­Dawley rats and the third passage cells were induced by 5­azacytidine (5­aza). Following successful induction, the cells were trypsinized and suspended at a density of 1x109/ml. Then, the cell suspension was added to the PLGA scaffold and cultured for 14 days. The morphological changes of BMMSCs were observed using phase contrast microscopy. Immunofluorescence staining was used to identify the cardiomyocyte­like cells. Hematoxylin and eosin (H&E) and immunohistochemical staining were used to observe the morphology of the engineered myocardial tissues. The cell adhesion rates and scanning electron microscopy were used to observe the compatibility of the cardiomyocyte­like cells and PLGA. Transmission electron microscopy was used to view the ultrastructure of the engineered myocardial tissues. BMMSCs in primary culture presented round or short spindle cell morphologies. Following induction by 5­aza, the cells exhibited a long spindle shape and a parallel arrangement. Analysis of the cell adhesion rates demonstrated that the majority of the cardiomyocyte­like cells had adhered to the PLGA scaffolds at 24 h. H&E staining suggested that the cardiomyocyte­like cells with spindle nuclei were evenly distributed in the PLGA scaffold. Immunofluorescence staining revealed that the cardiomyocyte­like cells were positive for cardiac troponin I. Scanning electron microscopy demonstrated that the inoculated cells were well attached to the PLGA scaffold. Transmission electron microscopy indicated that the engineered myocardial tissues contained well­arranged myofilaments, desmosomes, gap junction and Z line­like structures. The present study successfully constructed engineered myocardial tissues in vitro with a PLGA polymer and cardiomyocyte­like cells derived from BMMSCs, which are likely to share various structural similarities with the original heart tissue.


Assuntos
Células-Tronco Mesenquimais/citologia , Miócitos Cardíacos/citologia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Engenharia Tecidual/métodos , Tecidos Suporte/química , Animais , Diferenciação Celular , Células Cultivadas , Masculino , Miocárdio/citologia , Ratos Sprague-Dawley
13.
Nat Commun ; 10(1): 3123, 2019 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-31311935

RESUMO

Since both myocardium and vasculature in the heart are excessively damaged following myocardial infarction (MI), therapeutic strategies for treating MI hearts should concurrently target both so as to achieve true cardiac repair. Here we demonstrate a concomitant method that exploits the advantages of cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) and human mesenchymal stem cell-loaded patch (hMSC-PA) to amplify cardiac repair in a rat MI model. Epicardially implanted hMSC-PA provide a complimentary microenvironment which enhances vascular regeneration through prolonged secretion of paracrine factors, but more importantly it significantly improves the retention and engraftment of intramyocardially injected hiPSC-CMs which ultimately restore the cardiac function. Notably, the majority of injected hiPSC-CMs display adult CMs like morphology suggesting that the secretomic milieu of hMSC-PA constitutes pleiotropic effects in vivo. We provide compelling evidence that this dual approach can be a promising means to enhance cardiac repair on MI hearts.


Assuntos
Coração/fisiologia , Transplante de Células-Tronco Mesenquimais/métodos , Infarto do Miocárdio/terapia , Miócitos Cardíacos/transplante , Regeneração , Animais , Diferenciação Celular , Células Cultivadas , Modelos Animais de Doenças , Humanos , Células-Tronco Pluripotentes Induzidas/fisiologia , Injeções Intralesionais , Masculino , Infarto do Miocárdio/etiologia , Infarto do Miocárdio/patologia , Miocárdio/citologia , Miocárdio/patologia , Miócitos Cardíacos/fisiologia , Ratos , Ratos Endogâmicos F344 , Resultado do Tratamento
15.
Eur J Pharmacol ; 858: 172467, 2019 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-31216443

RESUMO

The present study investigates the cardioprotective effect of ß-caryophyllene against doxorubicin-induced acute cardiotoxicity in rats. Doxorubicin (12.5 mg/kg) and ß-caryophyllene (25, 50 or 100 mg/kg) were administered intraperitoneally to male albino Wistar rats. Doxorubicin-treated rats showed elevated levels of creatine kinase-MB in serum and oxidative stress in the myocardium as evidenced by decreased superoxide dismutase, catalase and glutathione with a concomitant rise in malondialdehyde levels. Doxorubicin also induced pro-inflammatory cytokines release following activation of the nuclear factor kappa-B and elevated expressions of inducible nitric oxide synthase and cyclooxygenase-2 in the myocardium. Additionally, doxorubicin also increased expression of γ-H2AX, a marker of DNA damage as well as increased expression of proapoptotic (Bax, p53, and active caspase-3) proteins along with the decreased expression of anti-apoptotic protein, Bcl2 in the myocardium. The histological and ultrastructural studies further revealed edema, inflammation and structural degeneration of cardiomyocytes following doxorubicin injection. However, treatment with ß-caryophyllene showed significant cardioprotective effects as evidenced by favorable improvement of biochemical and molecular parameters along with remarkable preservation of cardiomyocytes in histological and ultrastructural studies. Results of the present study demonstrate that ß-caryophyllene has potential to protect heart against doxorubicin-induced acute cardiotoxicity in rats. Moreover, the antioxidant and free radical scavenging properties of ß-caryophyllene was confirmed by in vitro assays. Provided the anticancer and chemosensitizing properties of ß-caryophyllene, the cardioprotective effects of ß-caryophyllene are suggestive of its multiple properties that provides an additional basis of its possible therapeutic application in chemotherapy-associated cardiotoxicity.


Assuntos
Apoptose/efeitos dos fármacos , Dieta , Doxorrubicina/toxicidade , Miocárdio/citologia , Miocárdio/metabolismo , Estresse Oxidativo/efeitos dos fármacos , /farmacologia , Animais , Antioxidantes/metabolismo , Peso Corporal/efeitos dos fármacos , Citocinas/metabolismo , Citoproteção/efeitos dos fármacos , Dano ao DNA , Relação Dose-Resposta a Droga , Radical Hidroxila/metabolismo , Inflamação/tratamento farmacológico , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Miocárdio/patologia , NF-kappa B/metabolismo , Ratos , Ratos Wistar , Superóxidos/metabolismo
16.
J Toxicol Sci ; 44(6): 425-433, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31168029

RESUMO

Cardiac fibroblasts (CFs) could be activated after myocardial infarction (MI). Thus, it is necessary to explore effective drugs to suppress the activation of CFs following MI. This study was designed to investigate the impacts of ellagic acid on CFs and the underlying mechanisms. The expression of histone deacetylases (HDACs) and fibrosis-related genes was detected by qRT-PCR and western blot. The Masson's Trichrome Staining assay was used to evaluate the area of cardiac fibrosis. The proliferation and migration of CFs were measured by CCK8 Kit and Transwell assay, respectively. Our results showed that ellagic acid significantly reduced protein expression of HDAC1, mRNA expression of collagen I, collagen III, MMP-2 and MMP-9 and the area of cardiac fibrosis in MI rats. In Ang II-stimulated CFs, ellagic acid (60 µmol/L) decreased the protein expression of HDAC1, collagen I, collagen III, MMP-2 and MMP-9, and inhibited cell proliferation and migration. Further, HDAC1 over-expression reversed the inhibitor effects of ellagic acid on proteins expression (collagen I, collagen III, MMP-2 and MMP-9) and proliferation and migration of CFs. The present results suggested that ellagic acid suppressed proliferation and migration of CFs by down-regulating expression of HDAC1.


Assuntos
Cardiotônicos/farmacologia , Ácido Elágico/farmacologia , Fibroblastos/efeitos dos fármacos , Angiotensina II , Animais , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Colágeno Tipo I/genética , Colágeno Tipo III/genética , Regulação para Baixo , Fibroblastos/fisiologia , Regulação da Expressão Gênica/efeitos dos fármacos , Histona Desacetilase 1/genética , Masculino , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , Miocárdio/citologia , Ratos Sprague-Dawley
17.
J Pharmacol Exp Ther ; 370(2): 308-317, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31160469

RESUMO

Protamine sulfate (PS) is widely used in heart surgery as an antidote for heparin, albeit its pharmacological effects are not fully understood and applications are often accompanied by unwanted side effects. Here we show the effect of PS on mitochondrial bioenergetics profile resulting in mitochondrial reactive oxygen species (ROS) production. Polarographic measurements were performed in parallel to membrane potential and ROS measurements by FACS analyzer using tetramethylrhodamine ethyl ester and MitoSOX fluorescent dyes, respectively. PS inhibited intact rat heart mitochondrial respiration (stimulated by ADP) to 76% (P < 0.001) from the baseline of 51.6 ± 6.9 to 12.4 ± 2.3 nmol O2⋅min-1⋅ml-1 The same effect was found when respiration was inhibited by antimycin A (101.0 ± 8.9 vs. 38.0 ± 9.9 nmol O2 ⋅min-1⋅ml-1, P < 0.001) and later stimulated by substrates of cytochrome oxidase (CytOx) i.e., ascorbate and tetramethyl phenylene diamine, suggesting that PS exerted its effect through inhibition of CytOx activity. Furthermore, the inhibition of mitochondrial respiration by PS was concentration dependent and accompanied by hyperpolarization of the mitochondrial membrane potential (Δψ m), i.e., 18% increase at 50 µg/ml and an additional 3.3% increase at 250 µg/ml PS compared with control. This effect was associated with a strong consequent increase in the production of ROS, i.e., 85% and 88.6% compared with control respectively. We propose that this excessive increase in ROS concentrations results in mitochondrial dysfunction and thus might relate to the "protamine reaction," contributing to the development of various cardiovascular adverse effects.


Assuntos
Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Protaminas/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Animais , Respiração Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Miocárdio/citologia , Miocárdio/metabolismo , Ratos , Ratos Wistar
18.
Folia Biol (Praha) ; 65(1): 36-42, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31171080

RESUMO

Hypothermic incubation of chicken eggs leads to smaller embryos with enlarged hearts, originally described as hypertrophic. Over the years, however, accumulated evidence suggested that hyperplasia, rather than hypertrophy, is the predominant mechanism of cardiac growth during the prenatal period. We have thus set to re-evaluate the hypothermia model to precise the exact cellular mechanism behind cardiac enlargement. Fertilized chicken eggs were incubated at either 37.5 °C (normothermia) or 33.5 °C from embryonic day (ED) 13 onward (hypothermia). Sampling was performed at ED17, at which point wet embryo and heart weight were recorded, and the hearts were submitted to histological examination. In agreement with previous results, the hypothermic embryos were 29% smaller and had hearts 18% larger, translating into a 67% increase in the heart to body weight ratio (P < 0.05 for all parameters). The cell size was essentially the same between control and hypothermic hearts in all regions analysed. Likewise, there was no significant relationship between the cell size and heart weight; however, in the hypothermic hearts, there was a trend showing positive correlation between cell sizes in different cardiac regions and heart weight. Proliferation rate, determined on the basis of anti-phosphohistone H3 immunofluorescence, showed an overall increase in the hypothermic group, reaching statistical significance (P = 0.02, t-test) in the right ventricle. The proliferation rate was similar among different regions of the same heart. However, the correlation between the proliferation rate and heart weight was only small (r2 = 0.007 and r2 = 0.234 for the normothermic and hypothermic group, respectively). We thus conclude that hyperplasia is the predominant response mechanism in this volume-overload model; mechanistically, decreased heart rate at lower temperature increases the end-diastolic and stroke volume, minimizing the drop in cardiac output through the Frank- Starling mechanism.


Assuntos
Hiperplasia/metabolismo , Hipertrofia/fisiopatologia , Hipotermia Induzida/métodos , Animais , Proliferação de Células/fisiologia , Embrião de Galinha , Imunofluorescência , Hipertrofia/metabolismo , Miocárdio/citologia , Miocárdio/metabolismo , Miócitos Cardíacos , Tamanho do Órgão/fisiologia , Volume Sistólico/fisiologia
19.
J Microencapsul ; 36(3): 267-277, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31195912

RESUMO

Aim: The aim of this study was to evaluate the formulation of a synthetic IGF-1 (pIGF-1) in PLGA microparticles (MP). Methods: Poly (lactic-co-glycolic acid) (PLGA) MPs loaded with pIGF-1 were prepared, characterised and evaluated using double emulsion solvent evaporation method. Results: Spherical MPs showed an average particle size of 2 µm, encapsulation efficiency (EE) of 67% and 50% degradation over 15 days. With a view to enhancing retention in the myocardium, the MP formulation was encapsulated in a cross-linked hyaluronic acid hydrogel. pIGF-1 released from MPs and from MPs suspended in hyaluronic acid hydrogel remained bioactive, determined by a significant increase in cellular proliferation of c-kit+ cells. Conclusion: This formulation has potential for loco-regional delivery to damaged myocardium to promote the survival of cardiomyocytes.


Assuntos
Portadores de Fármacos/química , Fator de Crescimento Insulin-Like I/administração & dosagem , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Ácido Hialurônico/química , Hidrogéis/química , Fator de Crescimento Insulin-Like I/farmacologia , Miocárdio/citologia , Tamanho da Partícula , Ratos
20.
Oxid Med Cell Longev ; 2019: 4273261, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31178959

RESUMO

Oxidative stress plays a major part in myocardial reperfusion injury. Cul4a is the core protein of CRLs E3 ubiquitin ligase complex; while it is known that Cul4a is responsible for various cancers, its role in cardiac function remains unclear. Hence, we have shown the protective function of Cul4a and its protection mechanism in oxidative stress-induced H9c2 cardiomyocyte apoptosis. Here, oxidative stress was induced by hydrogen peroxide (H2O2), CCK-8 assay and flow cytometry were used to analyze cell viability and apoptosis rate, western blot and immunofluorescence were used to quantitatively analyze the expression of protein, ROS fluorescence kit was used to detect reactive oxygen species (ROS) formation, and coimmunoprecipitation was used to identify protein interaction. In the results, it was found that Cul4a was involved in oxidative stress-induced H9c2 cell apoptosis and could inhibit H2O2-induced ROS generation and H9c2 cell apoptosis. Furthermore, we identified that when combining with PARP1, Cul4a could reduce its expression, and the interaction was enhanced under oxidative stress. In conclusion, our results indicate that Cul4a is a new protective factor involved in oxidative stress-induced cardiomyocyte injury and functions by tying and decreasing overactivated PARP1.


Assuntos
Proteínas Culina/metabolismo , Miocárdio/citologia , Miocárdio/metabolismo , Estresse Oxidativo/fisiologia , Poli(ADP-Ribose) Polimerase-1/metabolismo , Animais , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Proteínas Culina/genética , Peróxido de Hidrogênio/farmacologia , Poli(ADP-Ribose) Polimerase-1/genética , Ratos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Transfecção
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