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1.
Biol Trace Elem Res ; 200(1): 308-317, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33634365

RESUMO

Aluminum exposure can mediate either acute toxicity or chronic toxicity. Aluminum exerts toxic effects on the cardiovascular system, but there are few studies on its related mechanisms. In this study, we investigated the molecular mechanism of aluminum-induced oxidative damage and apoptosis in rat cardiomyocytes. Thirty-two male Wistar rats were randomly divided into four groups, including the control group (GC), low-dose group of aluminum exposure (GL), medium-dose group (GM), and high-dose group (GH), with eight rats in each group. The GL, GM, and GH groups were given 5, 10, and 20 mg/(kg·d) of AlCl3 solution by intraperitoneal injection, and the GC group received intraperitoneal injection of the same volume of normal saline (2 ml/rat/day), 5 times a week for 28 days. At the end of the experiment, the levels of aluminum, malondialdehyde (MDA), plasma lactate dehydrogenase (LDH), creatine kinase (CK), creatine kinase isoenzyme (CKMB), and alpha-hydroxybutyrate dehydrogenase (HBDH) were measured. The pathological changes of myocardium were observed by H&E staining. The apoptosis of cardiomyocytes was detected by TUNEL staining, and the expression of apoptosis-related proteins was determined by western blot. The results showed that the levels of CKMB and HBDH in the GM and GH groups were significantly higher than those in the GC group (P < 0.05). The content of aluminum in the myocardium and serum of the aluminum exposure groups was significantly higher than that of the GC group (P < 0.05). The level of MDA in the GM and GH groups was significantly higher than that in the GC group (P < 0.05). The pathological results showed that vacuolated and hypertrophied cardiomyocytes were found in aluminum exposure groups, especially in the GM and GH groups. The TUNEL staining showed that the apoptosis rate of the aluminum exposure groups was considerably higher than that of the GC group (P < 0.05). Western blot showed that the expression of Bcl-2, an anti-apoptotic protein, in cardiomyocytes of aluminum exposure groups was lower than that of the GC group (P < 0.05), while the levels of Bax and caspase-3 in the cardiomyocytes of the GM and GH groups were higher than those of the GC group (P < 0.05). The experimental results showed that aluminum could accumulate in myocardial tissues and cause damage to cardiomyocytes. It could induce oxidative stress damage by increasing the content of MDA in cardiomyocytes and trigger cardiomyocyte apoptosis by activating the pro-apoptotic proteins caspase-3 and Bax and reducing the anti-apoptotic protein Bcl-2.


Assuntos
Alumínio , Miócitos Cardíacos , Alumínio/metabolismo , Alumínio/toxicidade , Animais , Apoptose , Masculino , Miócitos Cardíacos/metabolismo , Estresse Oxidativo , Ratos , Ratos Wistar
2.
Biosens Bioelectron ; 195: 113570, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34455143

RESUMO

This paper proposes a new non-invasive, low-cost, and fully automated platform to quantitatively analyze dynamics of human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) at the single-cell level by holographic image-based tracking for cardiotoxicity screening. A dense Farneback optical flow method and holographic imaging informatics were combined to characterize the contractile motion of a single CM, which obviates the need for costly equipment to monitor a CM's mechanical beat activity. The reliability of the proposed platform was tested by single-cell motion characterization, synchronization analysis, motion speed measurement of fixed CMs versus live CMs, and noise sensitivity. The applicability of the motion characterization method was tested to determine the pharmacological effects of two cardiovascular drugs, isoprenaline (166 nM) and E-4031 (500 µM). The experiments were done using single CMs and multiple cells, and the results were compared to control conditions. Cardiomyocytes responded to isoprenaline by increasing the action potential (AP) speed and shortening the resting period, thus increasing the beat frequency. In the presence of E-4031, the AP speed was decreased, and the resting period was prolonged, thus decreasing the beat frequency. The findings offer insights into single hiPS-CMs' contractile motion and a deep understanding of their kinetics at the single-cell level for cardiotoxicity screening.


Assuntos
Técnicas Biossensoriais , Células-Tronco Pluripotentes Induzidas , Cardiotoxicidade , Células Cultivadas , Humanos , Miócitos Cardíacos , Reprodutibilidade dos Testes
3.
Zhongguo Zhong Yao Za Zhi ; 46(19): 5064-5071, 2021 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-34738402

RESUMO

The present study investigated the effects of chikusetsu saponin Ⅳa(CHS Ⅳa) on isoproterenol(ISO)-induced myocardial hypertrophy in rats and explored the underlying molecular mechanism. ISO was applied to establish a rat model of myocardial hypertrophy, and CHS Ⅳa(5 and 15 mg·kg~(-1)·d~(-1)) was used for intervention. The tail artery blood pressure was measured. Cardiac ultrasound examination was performed. The ratio of heart weight to body weight(HW/BW) was calculated. Morphological changes in the myocardial tissue were observed by HE staining. Collagen deposition in the myocardial tissue was observed by Masson staining. The mRNA expression of myocardial hypertrophy indicators(ANP and BNP), autophagy-related genes(Atg5, P62 and beclin1), and miR199 a-5 p was detected by qRT-PCR. Atg5 protein expression was detected by Western blot. The results showed that the model group exhibited increased tail artery blood pressure and HW/BW ratio, thickened left ventricular myocardium, enlarged myocardial cells, disordered myocardial fibers with widened interstitium, and a large amount of collagen aggregating around the extracellular matrix and blood vessels. ANP and BNP were largely expressed. Moreover, P62 expression was up-regulated, while beclin1 expression was down-regulated. After intervention by CHS Ⅳa at different doses, myocardial hypertrophy was ameliorated and autophagy activity in the myocardial tissue was enhanced. Meanwhile, miR199 a-5 p expression declined and Atg5 expression increased. As predicted by bioinformatics, Atg5 was a target gene of miR199 a-5 p. CHS Ⅳa was capable of preventing myocardial hypertrophy by regulating autophagy of myocardial cells through the miR-199 a-5 p/Atg5 signaling pathway.


Assuntos
Ácido Oleanólico , Saponinas , Animais , Cardiomegalia/induzido quimicamente , Cardiomegalia/tratamento farmacológico , Cardiomegalia/genética , Isoproterenol , Miocárdio , Miócitos Cardíacos , Ácido Oleanólico/análogos & derivados , Ratos , Saponinas/farmacologia
4.
Zhongguo Zhong Yao Za Zhi ; 46(20): 5210-5217, 2021 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-34738421

RESUMO

Pyroptosis is a pro-inflammatory programmed cell death, and its role in cardiac inflammatory response has become a hot topic. The activation of nucleotide binding oligomerization domain like receptor protein 3(NLRP3) inflammasome is an important mechanism for pyroptosis induced by cysteinyl aspartate specific proteinase 1(caspase-1). The existing studies have shown that cardiomyocyte pyroptosis participates in the pathogenesis of different cardiovascular diseases and the NLRP3 inflammasome-mediated cardiomyocyte pyroptosis has been most widely studied. Also, the intervention in NLRP3 inflammasome activation and cardiomyocyte pyroptosis contributes to ameliorating myocardial injury, which may be the main mechanism of many traditional Chinese medicines in exerting the cardio-protective effects. Therefore, this paper reviewed the studies on cardiomyocyte pyroptosis mediated by NLRP3 inflammasome and put forward the importance of exploring traditional Chinese medicine intervention in the activation of NLRP3 inflammasome.


Assuntos
Inflamassomos , Piroptose , Medicina Tradicional Chinesa , Miócitos Cardíacos , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética
5.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 37(5): 548-554, 2021 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-34816671

RESUMO

Objective: To investigate the mechanisms of dezocine on regulating H9C2 oxidative stress and apoptosis of rat cardiac myocytes induced by hypoxia-reoxygenation(H/R) by regulating the expressions of microRNA-7a- 5p(miR-7a-5p)/ubiquitin E3 ligase tripartite motif 10(TRIM10). Methods: H9C2 cells were divided into control group (cultured normally), H/R group (treated with hypoxia for 3 h and then reoxygenation for 4 h), different doses of dezocine intervention group (H9c2 cells were pretreated with dezocine at the concentrations of 10-7, 10-6 and 10-5 mmol/L for 24 h, and then treated with H/R), H/R+miR-7a-5p group (H9C2 cells were transfected with miR-7a-5p mimics and then treated with H/R), H/R+miR-NC group (H9C2 cells were transfected with miR-NC and then treated with H/R), H/R+Dezocine+anti-miR-7a-5p group (H9c2 cells transfected with anti-miR-7a-5p were pretreated with 10-5 mmol/L dezocine for 24 h, and then treated with H/R), H/R+dezocine+ anti-miR-NC Group (H9c2 cells transfected with anti-miR-NC were pretreated with 10-5 mmol/L dezocine for 24 h, and then treated with H/R). Each group of cells was set with 3 replicate wells, and the experiment was repeated 3 times. The content of malondialdehyde(MDA) and activity of superoxide dismutase(SOD) and glutathione peroxidas(GSH-Px) were detected by the enzyme-linked immunosorbent assay. The cells apoptosis was detected by flow cytometry. The protein expressions of B-cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax) and TRIM10 were detected by Western blot, and the expressions of miR-7a-5p and TRIM10 mRNA were detected by real-time quantitative PCR(RT-qPCR). The double luciferase reporter gene experiment was used to verify the regulatory relationship between miR-7a-5p and TRIM10. Results: Compared with the control group, the MDA content, apoptosis rate, the expression of Bax protein, and the expression of TRIM10 mRNA and protein in the H/R group were all increased (P<0.05), while the activities of SOD and GSH-Px, and the expressions of Bcl-2 protein and miR-7a-5p were all decreased (P<0.05). Compared with the H/R group, the MDA content, apoptosis rate, the expression of Bax protein, and the expression of TRIM10 mRNA and protein in the different doses of dezocine intervention group were decreased (P<0.05), while the activities of SOD and GSH-Px, and the expressions of Bcl-2 protein and miR-7a-5p were all increased (P<0.05), and there were significant differences in each index between the different doses of dezocine intervention groups (P< 0.05). Compared with the H/R+miR-NC group, the MDA content, apoptosis rate, the protein expressions of Bax and TRIM10 in the H/R+miR-7a-5p group were decreased (P<0.05), while the activities of SOD and GSH-Px, and the expression of Bcl-2 protein were all increased (P<0.05). Compared with the H/R+dezocine+anti- miR-NC group, the MDA content, apoptosis rate, the protein expressions of Bax and TRIM10 in the H/R+dezocine+anti-miR-7a-5p group were all increased (P<0.05), while the activities of SOD and GSH-Px, and the expression of Bcl-2 protein were all decreased (P<0.05). Conclusion: Dezocine can reduce oxidative stress and apoptosis of rat cardiomyocytes H9C2 induced by H/R, which may play a role in regulating the miR-7a-5p / TRIM10 axis.


Assuntos
MicroRNAs , Traumatismo por Reperfusão Miocárdica , Animais , Apoptose , Compostos Bicíclicos Heterocíclicos com Pontes , Hipóxia , MicroRNAs/genética , Miócitos Cardíacos , Ratos , Tetra-Hidronaftalenos
6.
Nihon Yakurigaku Zasshi ; 156(6): 346-350, 2021.
Artigo em Japonês | MEDLINE | ID: mdl-34719567

RESUMO

Intracellular Ca2+ plays pivotal roles in cardiac contraction by mediating excitation-contraction (EC) coupling and progression of hypertrophy in the heart. Ample evidence suggests that mechanism of EC coupling in immature hearts are different from those in the adults because of the structural immaturity of the sarcoplasmic reticulum (SR) intracellular Ca2+ store and the different expression of Ca2+-regulatory proteins. However, the detailed molecular mechanism is not completely understood. In the present study, we identified neuronal Ca2+ sensor-1 (NCS-1), an EF-hand Ca2+ binding protein that is important for neuronal functions, also functions as a novel regulator of EC coupling in young hearts. We found that NCS-1 is highly expressed in immature hearts, and its deletion decreased their contractile functions as well as intracellular Ca2+ signals. NCS-1 enhances Ca2+ signals mainly by promoting the Inositol 1,4,5-Trisphosphate receptor (IP3R) function, followed by Ca2+/Calmodulin-dependent Protein Kinase II (CaMKII) signaling, which results in a large increase in the SR Ca2+ content that enhances SR-dependent EC coupling. In addition, NCS-1 expression increases in the early stages of hypertrophy and promotes progression of hypertrophy at least in part through IP3R-dependent elevation of nuclear Ca2+ signaling. Our results reveal a previously unrecognized mechanism of EC coupling in young heart and the progression of cardiac hypertrophy. Furthermore, we found that NCS-1 contributes to stress tolerance in cardiomyocytes via activation of mitochondrial detoxification pathways. We propose that the proteins involved in NCS-1-mediated Ca2+ signaling can be novel therapeutic targets for cardiac diseases, especially in immature hearts.


Assuntos
Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina , Retículo Sarcoplasmático , Cálcio/metabolismo , Sinalização do Cálcio , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Miócitos Cardíacos , Retículo Sarcoplasmático/metabolismo , Transdução de Sinais
7.
Nihon Yakurigaku Zasshi ; 156(6): 351-354, 2021.
Artigo em Japonês | MEDLINE | ID: mdl-34719568

RESUMO

Heart failure is an important cause of death of children. Especially, overt one within the preweaning period is fulminant and severe. However, there are no drugs with evidence for it. We recently found that angiotensin II (AngII) activates L-type Ca2+ channels through AT1 receptors (AT1R) and ß-arrestin 2 in murine cardiac myocytes only in the preweaning period, indicating that AT1R/ß-arrestin 2 pathway mediates positive inotropic effects before weaning. Indeed, ß-arrestin-bias AT1R agonist (BBA), TRV027 caused significant long-lasting positive inotropic effects in preweaning mice without increasing serum aldosterone concentrations or inducing tachycardia, arrhythmias, increased cardiac oxygen consumption, and reactive oxygen species generation. TRV027 increased the peak amplitude of twitch Ca2+ transients not only in preweaning mouse cardiac myocytes but in human iPS cell-derived cardiac myocytes exhibiting the fetal to neonatal phenotype. Moreover, TRV027 also increased contraction of the compromised heart of the model knock-in mice mimicking human congenital dilated cardiomyopathy. Although ~80% of these mice died before weaning, TRV027 significantly increased their survival rate. TRV027 did not cause any obvious adverse effects on their preweaning wildtype littermates. Thus, we reason in this review that BBA can be important therapeutics for preweaning heart failure.


Assuntos
Insuficiência Cardíaca , Receptor Tipo 1 de Angiotensina , Angiotensina II , Animais , Insuficiência Cardíaca/tratamento farmacológico , Camundongos , Miócitos Cardíacos , Receptores de Angiotensina
8.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 37(6): 699-704, 2021 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-34821109

RESUMO

Objective: To establish a stable, rapid and improved method for isolation and culture of primary cardiomyocytes from neonatal rats. Methods: Ventricular tissues from neonatal SD rats were digested with 0.12% collagenase Ⅱ, and then subjected to Percoll density gradient centrifugation. The original cardiomyocytes were cultured in modified DMEM/F12 containing 5% horse serum and 5-bromodeoxyuracil(5-BrdU) in vitro for further purification, and medium was changed to normal high glucose DMEM with 10% FBS the next day. The difference between the improved method and traditional differential attachment one used for isolation and culture of primary cardiomyocytes was compared. Results: Cardiacmyocytes obtained through the improved method grew well. 24 hours after plating, most cells adhered to the dishes, with shapes looked triangular, fusiform or irregular, and some of them showed spontaneously contract at a frequency varying from 10~30 times/min. After 48 h culture, the cardiomyocytes became longer and stretched out pseudopodia. Some cells showed synchronous beats with the frequency close to 50~80 times/min. 72 hours later, cardiomyocytes were interwoven into a network in chrysanthemum patterns, and spontaneous beats tended to be more synchronous, with a frequency of 80-100 times/min. After 96 h, cells gathered into clusters as islands, with synchronous beat at a frequency of around 100~120 times/min. All cardiomyocytes were in good condition within one week. Yields((1.17±0.15)×106 vs (1.21±0.22)×106,P>0.05)and survival rate of primary cardiomyocytes obtained by the improved method was comparable to that gained using traditional differential attachment way (93.3%±1.4% vs 92.2%±0.7%, P>0.05), but the purity of primary cardiomyocytes obtained through the improved method was much higher (94.7%±2.1% vs 89.5%±1.3%, P<0.05), while with less time consuming ((3.1±0.4)h vs (4.3±0.3)h, P<0.01). Conclusion: This improved method is an ideal and simple method for the isolation and culture of primary cardiomyocytes with shorter time-consuming, high purity, intact structure and function, and with great repeatability and stability.


Assuntos
Ventrículos do Coração , Miócitos Cardíacos , Animais , Animais Recém-Nascidos , Células Cultivadas , Ratos , Ratos Sprague-Dawley
9.
Anticancer Res ; 41(11): 5355-5364, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34732405

RESUMO

Cardiotoxicity is a common side effect induced by cancer therapies, which increases the risk of long-term morbidity and mortality in cancer survivors. To date, the mechanism leading to this toxicity is still unclear, thus complicating cardiac safety assessment and predictive factor identification. The advances in technology, particularly regarding radiation therapy and constant development of novel antineoplastic agents, require urgent development of efficient preclinical models to detect drug cardiotoxicity. A myriad of empirical preclinical models have been used to investigate cardiotoxicity, though with limited success. Recently, multicellular spheroid models have gained attention by mimicking the in vivo microenvironment. The aim of this review is to focus on the most relevant preclinical models used to assess antineoplastic drug- and radiotherapy-related cardiotoxicities, with an overview on their current use. It also aims to discuss the possible directions of translational research in the cardio-oncology field.


Assuntos
Antineoplásicos , Cardiopatias/induzido quimicamente , Lesões por Radiação/etiologia , Pesquisa Médica Translacional , Animais , Técnicas Biossensoriais , Cardiotoxicidade , Linhagem Celular , Técnicas de Cocultura , Modelos Animais de Doenças , Cardiopatias/metabolismo , Cardiopatias/patologia , Cardiopatias/fisiopatologia , Humanos , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Lesões por Radiação/metabolismo , Lesões por Radiação/patologia , Lesões por Radiação/fisiopatologia , Radioterapia , Fatores de Risco , Especificidade da Espécie , Microambiente Tumoral
10.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 38(12): 1199-1203, 2021 Dec 10.
Artigo em Chinês | MEDLINE | ID: mdl-34839506

RESUMO

OBJECTIVE: To study the effect of down-regulating miR-488 targeting Jag1 on the injury of hypoxia-reoxygenation myocardial H9c2 cells. METHODS: A hypoxic-reoxygenated myocardial H9c2 cell injury model was constructed. miR-488 inhibitor was used to transfect the cells. CCK-8 method and flow cytometry were used to detect cell proliferation and apoptosis in each group. Lactate dehydrogenase (LDH), superoxide dismutase (SOD), malonaldehyde (MDA), catalase (CAT) levels were detected. Western blotting was used to detect the expression of Bcl-2 associated X Protein (Bax) and B cell lymphoma/lewkmia-2 (Bcl-2). Target genes of miR-488 were predicted, and a luciferase reporter system was used to verify the targeting relationship between the two. Myocardial H9c2 cells were co-transfected with miR-488 inhibitor and Jag1 siRNA, and treated with hypoxia and reoxygenation, cell proliferation, apoptosis, LDH, SOD, MDA, CAT levels, and Bax, Bcl-2 protein expression were detected. RESULTS: The expression of miR-488 in the hypoxia-reoxygenated myocardial H9c2 cells was increased, along with reduced cell proliferation, increased apoptosis, increased Bax protein expression, decreased Bcl-2 protein expression, increased MDA, decreased CAT and SOD, and increased LDH level in the supernatant of cell culture. When myocardial H9c2 cells were transfected with miR-488 inhibitor and treated with hypoxia and reoxygenation, the expression of miR-488 was decreased, along with increased cell proliferation, decreased apoptosis, decreased Bax protein expression, increased Bcl-2 protein expression, decreased MDA, increased CAT and SOD, and decreased LDH level in the supernatant of cell culture. Down-regulation of miR-488 could target and down-regulate Jag1 expression. And Jag1 siRNA could reverse the effect of miR-488 inhibitor on the proliferation, apoptosis, LDH, SOD, MDA, CAT levels and the expression of Bax and Bcl-2 of hypoxic-reoxygenated myocardial H9c2 cells. CONCLUSION: Down-regulating miR-488 targeted Jag1 can attenuate hypoxia-reoxygenation induced myocardial H9c2 cell injury.


Assuntos
MicroRNAs , Traumatismo por Reperfusão Miocárdica , Apoptose/genética , Regulação para Baixo , Humanos , Hipóxia/genética , Proteína Jagged-1/genética , MicroRNAs/genética , Miócitos Cardíacos
11.
Georgian Med News ; (318): 165-171, 2021 Sep.
Artigo em Russo | MEDLINE | ID: mdl-34628401

RESUMO

Over the past decades, there has been an active scientific search for drugs that can increase myocardial contractility and improve the course of heart failure. Omecamtiv Mecarbil, a drug from the group of cardiac myosin activators, heads the list of applicants for clinical use. The article presents the results of several randomized clinical trials which studied the efficacy and safety of Omecamtiv Mecarbil in heart failure: ATOMIC-AHF, COSMIC-HF and GALACTIC-HF. ATOMIC-AHF showed a tendency to reduce the risk of developing supraventricular and ventricular arrhythmias in heart failure. COSMIC-HF has proven the ability of Omecamtiv Mecarbil to improve the quality of life of patients with heart failure. GALACTIC-HF may be a turning point in the medical treatment of heart failure. For the first time, clinical evidence of the ability of the selective cardiac myosin activator Omecamtiv Mecarbil to improve myocardial contractile function, reduce the severity of symptoms of heart failure and reduce the risk of cardiovascular death was obtained.


Assuntos
Insuficiência Cardíaca , Miócitos Cardíacos , Insuficiência Cardíaca/tratamento farmacológico , Humanos , Miosinas , Qualidade de Vida , Volume Sistólico , Resultado do Tratamento , Ureia/análogos & derivados
12.
J Vis Exp ; (175)2021 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-34605811

RESUMO

The ability to study human cardiac development in health and disease is highly limited by the capacity to model the complexity of the human heart in vitro. Developing more efficient organ-like platforms that can model complex in vivo phenotypes, such as organoids and organs-on-a-chip, will enhance the ability to study human heart development and disease. This paper describes a protocol to generate highly complex human heart organoids (hHOs) by self-organization using human pluripotent stem cells and stepwise developmental pathway activation using small molecule inhibitors. Embryoid bodies (EBs) are generated in a 96-well plate with round-bottom, ultra-low attachment wells, facilitating suspension culture of individualized constructs. The EBs undergo differentiation into hHOs by a three-step Wnt signaling modulation strategy, which involves an initial Wnt pathway activation to induce cardiac mesoderm fate, a second step of Wnt inhibition to create definitive cardiac lineages, and a third Wnt activation step to induce proepicardial organ tissues. These steps, carried out in a 96-well format, are highly efficient, reproducible, and produce large amounts of organoids per run. Analysis by immunofluorescence imaging from day 3 to day 11 of differentiation reveals first and second heart field specifications and highly complex tissues inside hHOs at day 15, including myocardial tissue with regions of atrial and ventricular cardiomyocytes, as well as internal chambers lined with endocardial tissue. The organoids also exhibit an intricate vascular network throughout the structure and an external lining of epicardial tissue. From a functional standpoint, hHOs beat robustly and present normal calcium activity as determined by Fluo-4 live imaging. Overall, this protocol constitutes a solid platform for in vitro studies in human organ-like cardiac tissues.


Assuntos
Organoides , Células-Tronco Pluripotentes , Diferenciação Celular , Humanos , Mesoderma , Miócitos Cardíacos
13.
J Cardiothorac Surg ; 16(1): 301, 2021 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-34654440

RESUMO

BACKGROUND: This study sought to investigate the predictive value and regulatory mechanism of serum miR-499a-5p in sepsis-induced myocardial dysfunction (SIMD). METHODS: A total of 60 patients with sepsis and 60 healthy volunteers were enrolled in this study. The serum levels of miRNAs (miR-451, miR-378 and miR-499a-5p) were detected. Receiver operating characteristic curve and logistic regression analysis were used to evaluate the diagnostic and prognostic value of miR-499a-5p in SIMD patients. AC16 cells were used to establish SIMD model in vitro using lipopolysaccharide (LPS). An analysis was conducted for miR-499a-5p expression, cell viability, and the concentration of creatine kinase-MB isoform (CK-MB), brain natriuretic peptide (BNP), superoxide dismutase (SOD) and cytochrome C oxidase IV (COX IV). The downstream target of miR-499a-5p was verified. RESULTS: Our results revealed a poor expression of miR-499a-5p in the serum of SIMD patients, while no significant difference was evident for miR-451 and miR-378. The level of miR-499a-5p in the survival group was higher than the non-survival group. miR-499a-5p elicited good diagnostic and prognostic value for SIMD. Our findings revealed that miR-499a-5p was decreased significantly in LPS-treated cardiomyocytes. After overexpression of miR-499a-5p, the cell viability increased, and the concentrations of CK-MB and BNP were decreased, while the concentrations of SOD and COX IV were increased. EIF4E was validated as the target of miR-499a-5p. After overexpression of EIF4E, the cell viability was decreased and the concentrations of CK-MB and BNP were increased while the concentrations of SOD and COX IV were decreased. CONCLUSION: The level of miR-499a-5p is weak in SIMD patients. miR-499a-5p has a good diagnostic and prognostic value for SIMD by inhibiting EIF4E transcription.


Assuntos
Coração/fisiopatologia , MicroRNAs , Sepse , Creatina Quinase Forma MB , Humanos , MicroRNAs/genética , Miocárdio , Miócitos Cardíacos , Prognóstico , Sepse/diagnóstico
14.
Nat Commun ; 12(1): 5804, 2021 10 04.
Artigo em Inglês | MEDLINE | ID: mdl-34608155

RESUMO

During the last decade, cardiac optogenetics has turned into an essential tool for investigating cardiac function in general and for assessing functional interactions between different myocardial cell types in particular. To advance exploitation of the unique research opportunities offered by this method, we develop a panoramic opto-electrical measurement and stimulation (POEMS) system for mouse hearts. The core of the experimental platform is composed of 294 optical fibers and 64 electrodes that form a cup which embraces the entire ventricular surface of mouse hearts and enables straightforward 'drop&go' experimentation. The flexible assignment of fibers and electrodes to recording or stimulation tasks permits a precise tailoring of experiments to the specific requirements of individual optogenetic constructs thereby avoiding spectral congestion. Validation experiments with hearts from transgenic animals expressing the optogenetic voltage reporters ASAP1 and ArcLight-Q239 demonstrate concordance of simultaneously recorded panoramic optical and electrical activation maps. The feasibility of single fiber optical stimulation is proven with hearts expressing the optogenetic voltage actuator ReaChR. Adaptation of the POEMS system to larger hearts and incorporation of additional sensors can be achieved by redesigning the system-core accordingly.


Assuntos
Coração/fisiologia , Optogenética/métodos , Animais , Técnicas Eletrofisiológicas Cardíacas , Frequência Cardíaca , Potenciais da Membrana , Camundongos , Camundongos Transgênicos , Miócitos Cardíacos/fisiologia , Optogenética/instrumentação , Imagens com Corantes Sensíveis à Voltagem
15.
Lab Chip ; 21(20): 3899-3909, 2021 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-34636821

RESUMO

Human induced pluripotent stem (iPS) cell-derived cardiomyocytes are used for in vitro pharmacological and pathological studies worldwide. In particular, the functional assessment of cardiac tissues created from iPS cell-derived cardiomyocytes is expected to provide precise prediction of drug effects and thus streamline the process of drug development. However, the current format of electrophysiological and contractile assessment of cardiomyocytes on a rigid substrate is not appropriate for cardiac tissues that beat dynamically. Here, we show a novel simultaneous measurement system for contractile force and extracellular field potential of iPS cell-derived cardiac cell sheet-tissues using 500 nm-thick flexible electronic sheets. It was confirmed that the developed system is applicable for pharmacological studies and assessments of excitation-contraction coupling-related parameters, such as the electro-mechanical window. Our results indicate that flexible electronics with cardiac tissue engineering provide an advanced platform for drug development. This system will contribute to gaining new insight in pharmacological study of human cardiac function.


Assuntos
Células-Tronco Pluripotentes Induzidas , Diferenciação Celular , Eletrônica , Humanos , Miócitos Cardíacos , Engenharia Tecidual
16.
Int J Mol Sci ; 22(19)2021 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-34639171

RESUMO

Diabetes is a major risk factor for cardiovascular diseases, especially cardiomyopathy, a condition in which the smooth muscles of the heart become thick and rigid, affecting the functioning of cardiomyocytes, the contractile cells of the heart. Uncontrolled elevated glucose levels over time can result in oxidative stress, which could lead to inflammation and altered epigenetic mechanisms. In the current study, we investigated whether hyperglycemia can modify cardiac function by directly affecting these changes in cardiomyocytes. To evaluate the adverse effect of high glucose, we measured the levels of gap junction protein, connexin 43, which is responsible for modulating cardiac electric activities and Troponin I, a part of the troponin complex in the heart muscles, commonly used as cardiac markers of ischemic heart disease. AC16 human cardiomyocyte cells were used in this study. Under hyperglycemic conditions, these cells demonstrated altered levels of connexin 43 and Troponin-I after 24 h of exposure. We also examined hyperglycemia induced changes in epigenetic markers: H3K9me1, Sirtuin-1 (SIRT1), and histone deacetylase (HDAC)-2 as well as in inflammatory and stress-related mediators, such as heat shock protein (HSP)-60, receptor for advanced glycation end products (RAGE), toll-like receptor (TLR)-4, high mobility group box (HMGB)-1 and CXC chemokine receptor (CXCR)-4. Cardiomyocytes exposed to 25mM glucose resulted in the downregulation of HSP60 and SIRT1 after 48 h. We further examined that hyperglycemia mediated the decrease in the gap junction protein CX43, as well as CXC chemokine receptor CXCR4 which may affect the physiological functions of the cardiomyocytes when exposed to high glucose for 24 and 48 h. Upregulated expression of DNA-binding nuclear protein HMGB1, along with changes in histone methylation marker H3K9me1 have demonstrated hyperglycemia-induced damage to cardiomyocyte at 24 h of exposure. Our study established that 24 to 48 h of hyperglycemic exposure could stimulate stress-mediated inflammatory mediators in cardiomyocytes in vitro. These stress-related changes in hyperglycemia-induced cardiomyocytes may further initiate an increase in injury markers which eventually could alter the epigenetic processes. Therefore, epigenetic and inflammatory mechanisms in conjunction with alterations in a downstream signaling pathway could have a direct effect on the functionality of the cardiomyocytes exposed to high glucose during short and long-term exposures.


Assuntos
Biomarcadores/metabolismo , Epigênese Genética , Hiperglicemia/fisiopatologia , Mediadores da Inflamação/metabolismo , Miócitos Cardíacos/patologia , Estresse Fisiológico , Chaperonina 60/genética , Chaperonina 60/metabolismo , Conexina 43/genética , Conexina 43/metabolismo , Proteína HMGB1/genética , Proteína HMGB1/metabolismo , Histona Desacetilase 2/genética , Histona Desacetilase 2/metabolismo , Humanos , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Miócitos Cardíacos/metabolismo , Receptores CXCR4/genética , Receptores CXCR4/metabolismo , Sirtuína 1/genética , Sirtuína 1/metabolismo
17.
Biopreserv Biobank ; 19(5): 376-385, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34672722

RESUMO

Hypoxic-ischemic brain damage (HIBD) is a leading cause of fatality and neural system injury in neonates. This study aims to explore the effect of long noncoding RNA H19 on cardiomyocyte apoptosis in neonatal rats with HIBD. The neonatal rat model of HIBD was established. The cerebral infarction volume and apoptosis index of cardiomyocyte increased, while H19 expression decreased in neonatal rats with HIBD. After the lentivirus vector of overexpressed H19 was injected into neonatal rats with HIBD, the cardiomyocyte apoptosis was suppressed; levels of inflammatory factors and oxidative stress injury of myocardial tissues were reduced. The binding relationships between H19 and miR-149-5p, and miR-149-5p and leukemia inhibitory factor (LIF) were predicted by a bioinformatics website and verified using the dual-luciferase reporter gene assay. H19 competitively bound to miR-149-5p to upregulate LIF expression and activate the PI3K/Akt pathway. Moreover, a functional rescue experiment was carried out. Injection of Wortmannin reversed the inhibitory effect of H19 overexpression on cardiomyocyte apoptosis in neonatal rats with HIBD. It could be concluded that H19 competitively bound to miR-149-5p to upregulate LIF expression and activate the PI3K/Akt pathway, thus reducing cardiomyocyte apoptosis in neonatal rats with HIBD. This study may offer new insights for HIBD treatment.


Assuntos
MicroRNAs , RNA Longo não Codificante , Animais , Animais Recém-Nascidos , Apoptose , Encéfalo/metabolismo , Fator Inibidor de Leucemia , MicroRNAs/genética , Miócitos Cardíacos/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Longo não Codificante/genética , Ratos
18.
Int J Mol Sci ; 22(19)2021 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-34638793

RESUMO

Differentiation of pluripotent stem cells to cardiomyocytes is influenced by culture conditions including the extracellular matrices or similar synthetic scaffolds on which they are grown. However, the molecular mechanisms that link the scaffold with differentiation outcomes are not fully known. Here, we determined by immunofluorescence staining and mass spectrometry approaches that extracellular matrix (ECM) engagement by mouse pluripotent stem cells activates critical components of canonical wingless/integrated (Wnt) signaling pathways via kinases of the focal adhesion to drive cardiomyogenesis. These kinases were found to be differentially activated depending on type of ECM engaged. These outcomes begin to explain how varied ECM composition of in vivo tissues with development and in vitro model systems gives rise to different mature cell types, having broad practical applicability for the design of engineered tissues.


Assuntos
Diferenciação Celular , Matriz Extracelular/metabolismo , Adesões Focais/metabolismo , Modelos Cardiovasculares , Miócitos Cardíacos/metabolismo , Células-Tronco Pluripotentes/metabolismo , Via de Sinalização Wnt , Animais , Camundongos
19.
Gen Physiol Biophys ; 40(5): 419-426, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34602455

RESUMO

Hypertrophic cardiomyopathy (HCM) is a heterogeneous myocardial disease characterized by myocardial hypertrophy, myocardial mechanical and electrical activity obstacles. This study aimed to explore the relationship between YAP2 (Yes-associated protein 2) and HCM and clarify a signaling path about the pathogenesis of HCM. Our study confirms that YAP2 can promote myocardial cell hypertrophy at the molecular level (myocardial lineage cell H9C2), organ level (clinical specimens of human HCM), and an animal model (a mouse model of HCM with cardiac-specific transgenic and knockout YAP2). The detailed molecular mechanisms linking YAP2 to cardiomyocyte hypertrophy and HCM were investigated. This study proved that YAP2, as the final reaction factor in Hippo pathways, influences Akt1 activity to affect the downstream genes, which participate in the formation of HCM by promoting myocardial cell proliferation and cardiac hypertrophy.


Assuntos
Cardiomiopatia Hipertrófica , Animais , Modelos Animais de Doenças , Camundongos , Miocárdio , Miócitos Cardíacos , Transdução de Sinais
20.
Analyst ; 146(22): 6768-6779, 2021 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-34642716

RESUMO

Herein, we propose an array of gold (Au)-coated SU-8 cantilevers with microgrooves for improved maturation of cardiomyocytes and describe its applications to drug-induced cardiac toxicity tests. Firstly, we evaluated the effect of cell culture substrates such as polydimethylsiloxane (PDMS), polyimide (PI), and SU-8 on the cardiomyocyte's maturation. Among these, the SU-8 with microgroove structures exhibits improved cardiomyocyte maturation. Further, thin layers of graphene and Au are coated on SU-8 substrates and the effects of these materials on cardiomyocyte maturation are evaluated by analyzing the expression of proteins such as alpha-actinin, Connexin 43 (Cx43), and Vinculin. While both conductive materials enhanced protein expression when compared to bare SU-8, the Au-coated SU-8 substrates demonstrated superior cardiomyocyte maturation. The cantilever structure is constructed using microgroove patterned SU-8 with and without an Au coating. The Au-coated SU-8 cantilever showed maximum displacement of 17.6 ± 0.3 µm on day 21 compared to bare SU-8 (14.2 ± 0.4 µm) owing to improved cardiomyocytes maturation. Verapamil and quinidine are used to characterize drug-induced changes in the contraction characteristics of cardiomyocytes on bare and Au-coated SU-8 cantilevers. The relative contraction forces and beat rates changed according to the calcium and sodium channel related drugs. Matured cardiomyocytes are less influenced by the drugs compared to immature cardiomyocytes and showed reliable IC50 values. These results indicate that the proposed Au-coated SU-8 cantilever array could help improve the accuracy of toxicity screening results by allowing for the use of cardiomyocytes that have been matured on the drug screening platform.


Assuntos
Cardiotoxicidade , Preparações Farmacêuticas , Ouro/toxicidade , Humanos , Contração Miocárdica , Miócitos Cardíacos
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