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cardiomyocytes and flow cytometry

Hong-Yuan Zhang, Ting-Ting Zhang, Jeremiah Ong'achwa Machuki, Li-Juan Wu, Hong Sun
The present study was aimed to characterize the electrophysiology of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). IMR90-4 cells were induced to differentiate into cardiomyocytes by temporal modulation of regulators of canonical Wnt signaling. The protein expression of cardiac troponin T (cTnT) was detected by immunofluorescence staining and flow cytometry, and the differentiation rate of hiPSC-CMs was calculated. The action potentials (APs) of hiPSC-CMs were recorded by patch clamp and used to classify different types of cardiomyocytes...
June 25, 2018: Sheng Li Xue Bao: [Acta Physiologica Sinica]
Jie Yu, Hongwei Gao, Chuanhong Wu, Qiong-Ming Xu, Jin-Jian Lu, Xiuping Chen
Doxorubicin (DOX) is a widely used antineoplastic agent in clinics. However, its clinical application is largely limited by its cardiotoxicity. Diethyl blechnic (DB) is a novel compound isolated from Salvia miltiorrhiza Bunge. Here, we study the effect of DB on DOX-induced cardiotoxicity and its underlying mechanisms. Cellular viability was tested by 3-[-4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and protein level was evaluated by Western blotting. 5,5’,6,6’-tetrachloro-1,1’,3,3’-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) staining was performed to determine the mitochondrial membrane potential (MMP)...
June 19, 2018: International Journal of Molecular Sciences
David Brenière-Letuffe, Aya Domke-Shibamiya, Arne Hansen, Thomas Eschenhagen, Boris Fehse, Kristoffer Riecken, Justus Stenzig
BACKGROUND: Human induced pluripotent stem (iPS) cells have revolutionised research and spark hopes for future tissue replacement therapies. To obtain high cell numbers, iPS cells can be expanded indefinitely. However, as long-term expansion can compromise cell integrity and quality, we set out to assess potential reduction of clonal diversity by inherent growth imbalances. METHODS: Using red, green, blue marking as a lentiviral multi-colour clonal cell tracking technology, we marked three different iPS cell lines as well as three other cell lines, assigning a unique fluorescent colour to each cell at one point in culture...
June 18, 2018: Stem Cell Research & Therapy
Le Geng, Zidun Wang, Chang Cui, Yue Zhu, Jiaojiao Shi, Jiaxian Wang, Minglong Chen
BACKGROUND/AIMS: Heart failure induced by tachycardia, the most common arrhythmia, is frequently observed in clinical practice. This study was designed to investigate the underlying mechanisms. METHODS: Rapid electrical stimulation (RES) at a frequency of 3 Hz was applied on human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) for 7 days, with 8 h/day and 24 h/day set to represent short-term and long-term tachycardia, respectively. Age-matched hiPSC-CMs without electrical stimulation or with slow electrical stimulation (1 Hz) were set as no electrical stimulation (NES) control or low-frequency electrical stimulation (LES) control...
June 15, 2018: Cellular Physiology and Biochemistry
Andrew S Riching, Yuanbiao Zhao, Yingqiong Cao, Pilar Londono, Hongyan Xu, Kunhua Song
Trans-differentiation of one somatic cell type into another has enormous potential to model and treat human diseases. Previous studies have shown that mouse embryonic, dermal, and cardiac fibroblasts can be reprogrammed into functional induced-cardiomyocyte-like cells (iCMs) through overexpression of cardiogenic transcription factors including GATA4, Hand2, Mef2c, and Tbx5 both in vitro and in vivo. However, these previous studies have shown relatively low efficiency. In order to restore heart function following injury, mechanisms governing cardiac reprogramming must be elucidated to increase efficiency and maturation of iCMs...
June 3, 2018: Journal of Visualized Experiments: JoVE
Duomeng Yang, Junmin Xi, Yun Xing, Xiangxu Tang, Xiaomeng Dai, Kaiying Li, Hongmei Li, Xiuxiu Lv, Daxiang Lu, Huadong Wang
BACKGROUND: Neonatal rat ventricular myocytes (NRVMs) have proven to be an ideal research model for cardiac disease. However, the current methods to purify NRVMs have a limitation to obtain high purity. The purpose of this study was to develop a NRVM purification method by using superparamagnetic iron oxide particles (SIOP). METHODS: NRVMs were purified by using SIOP (SIOP group). The differential attachment with or without bromodeoxyuridine (BrdU) treatment served as control and BrdU groups, respectively...
June 3, 2018: International Journal of Cardiology
Zhengwen Zhou, Yue Zhang, Luning Lin, Jianmei Zhou
Apigenin, a flavonoid with multiple physiological and pharmacological activities, is associated with the prevention of cardiovascular diseases. The present study aimed to examine the roles and mechanisms of apigenin in the apoptosis of H9C2 rat cardiomyocytes, which were subjected to myocardial ischemia‑reperfusion (MI/R) injury. Cell viability, reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and cellular apoptosis were evaluated using cell counting kit‑8 assays and flow cytometry...
May 31, 2018: Molecular Medicine Reports
Ji-Gang He, Hong-Rong Li, Jin-Xiu Han, Bei-Bei Li, Dan Yan, Hong-Yuan Li, Ping Wang, Ying Luo
This study aimed to investigate whether exosomes secreted by mouse GATA-4-expressing bone marrow mesenchymal stem cells (BMSCs) could induce BMSC differentiation into myocyte precursors, decrease cardiomyocyte apoptosis, and improve cardiac function following myocardial infarction (MI). BMSCs were transduced with a lentivirus carrying a doxycycline (DOX)-inducible GATA-4 or control lentivirus, and secreted exosomes from these BMSCs were collected and co-cultured with BMSCs or cardiomyocytes under hypoxic and serum free conditions...
June 13, 2018: Scientific Reports
Lili Zhang, Zhiyan Wang, Di Wang, Jumo Zhu, Yi Wang
CD8+ T cells accumulate in the necrotic myocardium of acute myocardial infarction (AMI). It is unclear whether CD8+ CD28+ T cells, a specific subset of CD8+ T cells, contribute to myocardial injury. In this study, 92 consecutive patients with AMI and 28 healthy control subjects were enrolled. The frequency of CD8+ CD28+ T cells in peripheral blood samples was assayed by flow cytometry. Plasma cardiac troponin I (TNI) and left ventricular ejection fraction (LVEF) were determined. Long-term prognosis of the patients was evaluated by major adverse cardiac and cerebrovascular events (MACCE) over a 12-month follow-up period...
June 7, 2018: Molecular Immunology
Lei Qin, Wen Yang, Yao-Xin Wang, Zhen-Jun Wang, Chen-Chen Li, Man Li, Jie-Yun Liu
INTRODUCTION: Myocardial ischemia-reperfusion (I/R) injury affects millions of people worldwide and has a very high mortality rate. Since microRNA-497 (miR-497) has been found to be related with cardiomyocyte apoptosis, this study aimed to explore the effect of miR-497 by targeting Mfn2 in a mouse model of myocardial ischemia-reperfusion (I/R) injury. MATERIALS: BALB/c mice were modeled with I/R and some were injected with miR-497 agomir before I/R to observe whether miR-497 alleviates the injury that occurs as a result of I/R...
May 28, 2018: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
Hongwei Zhao, Xiaobei Zhang, Ying Zheng, Yuan Li, Xiaokun Wang, Nan Hu, Peng Zhou, Kaiyuan Wang
We aimed to evaluate the cardioprotective effect and mechanism of propofol in anthracycline-induced cardiomyocyte apoptosis. We selected the rat myocardial cell line, H9c2, and primary cardiomyocytes for in vitro study. The cardiomyocytes were treated with vehicle, Adriamycin® (ADM), propofol, or a combination of ADM and propofol. The proportion of apoptotic cells and the expression of miR-181a were detected by flow cytometry and real-time PCR, respectively. Luciferase assays were performed to explore the direct target gene of miR-181a...
2018: Oxidative Medicine and Cellular Longevity
Ling Wang, Rong Lin, Langtao Guo, Meiman Hong
The present study aimed to investigate whether pretreatment with rosuvastatin (RS) can provide cardioprotection in a myocardial ischemia/reperfusion (MI/R) model. The protective effect of RS on myocardial oxygen‑glucose deprivation/reperfusion (OGD/R) injury was also evaluated by upregulating peroxisome proliferator‑activated receptor‑γ (PPAR‑γ). In the present study, MI/R model was established and activities of superoxide dismutase (SOD), lactate dehydrogenase (LDH), creatine kinase‑muscle/brain (CK‑MB), malondialdehyde (MDA), and troponin I/T were measured...
May 23, 2018: Molecular Medicine Reports
Zhu-Xia Shen, Qing-Zhen Yang, Chao Li, Lin-Juan Du, Xue-Nan Sun, Yan Liu, Jian-Yong Sun, Hui-Hui Gu, Yu-Min Sun, Jun Wang, Sheng-Zhong Duan
BACKGROUND AND AIMS: Agonists of peroxisome proliferator-activated receptor gamma (Pparγ) have been demonstrated to reduce the risk of myocardial infarction (MI) in clinical trials and animal experiments. However, the cellular and molecular mechanisms are not completely understood. We aimed to reveal the functions of myeloid Pparγ in MI and explore the potential mechanisms in this study. METHODS: Myeloid Pparγ knockout (MPGKO) mice (n = 12) and control mice (n = 8) underwent coronary artery ligation to induce MI...
May 5, 2018: Atherosclerosis
Yifan Su, Hua Tian, Lijiang Wei, Guohui Fu, Ting Sun
Hypoxia-induced apoptosis plays an important role in cardiovascular diseases. Integrin β3 is one of the main integrin heterodimer receptors on the surface of cardiac myocytes. However, despite the important role that integrin β3 plays in the cardiovascular disease, its exact role in the hypoxia response remains unclear. Hence, in the present investigation we aimed to study the role of integrin β3 in hypoxia-induced apoptosis in H9C2 cells and primary rat myocardial cells. MTT assay, flow cytometry and TUNEL assay results showed that hypoxia inhibited cardiomyocyte proliferation and induced cardiomyocyte apoptosis...
May 23, 2018: Acta Biochimica et Biophysica Sinica
Juan Gao, Min Zhu, Rui-Feng Liu, Jian-Shu Zhang, Ming Xu
Background: MicroRNA-24 (miR-24) plays an important role in heart failure by reducing the efficiency of myocardial excitation-contraction coupling. Prolonged cardiac hypertrophy may lead to heart failure, but little is known about the role of miR-24 in cardiac hypertrophy. This study aimed to preliminarily investigate the function of miR-24 and its mechanisms in cardiac hypertrophy. Methods: Twelve Sprague-Dawley rats with a body weight of 50 ± 5 g were recruited and randomly divided into two groups: a transverse aortic constriction (TAC) group and a sham surgery group...
June 5, 2018: Chinese Medical Journal
Jingli Cao, Yingfeng Wu, Geming Liu, Zhenlong Li
Objective To investigate the role and molecular mechanism of brain-derived neurotrophic factor (BDNF) against the process of cardiomyocyte hypertrophy and apoptosis. Methods Cardiomyocyte hypertrophy were estabolished by angiotensin II (Ang II) in neonatal cardiomyocytes in vitro and incomplete ligature of abdominal aorta of SD rats in vivo. BDNF over-expressing recombinant vector pcDNA5-BDNF was transfected into cardiomyocytes by liposomes. Immunofluorescence staining was used to detect the effect of BDNF transfection on the surface area of myocardial cells...
March 2018: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
Jiang Rui, Shaohong Fang, Yongchen Wang, Bo Lv, Bo Yu, Shufeng Li
Bone marrow‑derived mesenchymal stem cells (BM‑MSCs) have been used in experimental research and clinical trials for heart function restoration and cardiomyocyte regeneration. However, due to a hostile microenvironment created by ischemia, hypoxia and pro‑inflammatory factors, the survival rate of implanted BM‑MSCs remains low. Therefore, strategies that can promote BM‑MSC survival and prevent apoptosis are required. Previous studies have reported that microRNA‑16 (miR‑16) can inhibit cell proliferation by targeting several proteins and signal pathway, not only by inducing apoptosis...
May 11, 2018: Molecular Medicine Reports
Chao Zhao, Yong Mei, Xufeng Chen, Lei Jiang, Yunfei Jiang, Xu Song, Hang Xiao, Jun Wang, Jingsong Zhang
Methamphetamine (METH) is a commonly abused psychostimulant that can induce severe neurotoxicity. Cardiovascular injury caused by METH has recently gained increasing attention; however, the underlying mechanisms remain unclear. As autophagy has been shown to be associated with cell injury, the association between autophagy and METH-mediated cell apoptosis was investigated in the present study. METH treatment significantly increased the expression of two key autophagy proteins, i.e., Beclin-1 and LC3-II, in the cardiomyocyte cell line H9C2...
May 12, 2018: Toxicology Letters
Jing Cheng, Qianfu Wu, Rong Lv, Li Huang, Banglong Xu, Xianbao Wang, Aihua Chen, Fei He
BACKGROUND/AIMS: The present study aimed to detect the expression of miR-449a and investigate the effect of miR-449a on cell injury in cardiomyocytes subjected to hypoxia/ reoxygenation (H/R) and its underlying mechanisms. METHODS: The expression of miR-449a was determined using reverse transcription-polymerase chain reaction in both neonatal rat ventricular myocytes and H9C2 cells. For gain-of-function and loss-of-function studies, H9C2 cells were transfected with either miR-449a mimics or miR-449a inhibitor...
May 7, 2018: Cellular Physiology and Biochemistry
Qiang Wang, Yunfa Xu, Ying Gao, Qi Wang
Cardiac hypertrophy is frequently accompanied by ischemic heart disease. Actinidia chinensis planch polysaccharide (ACP) is the main active compound from Actinidia chinensis planch. In the present study, a cardiac hypertrophy model was produced by treating cells with Angiotensin II (Ang II), which was used to investigate whether ACP protected against cardiac hypertrophy in vitro. It was demonstrated that ACP alleviated Ang II‑induced cardiac hypertrophy. In addition, pretreatment with ACP prior to hypoxic culture reduced the disruption of the mitochondrial membrane potential as investigated by flow cytometry...
May 3, 2018: Molecular Medicine Reports
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