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cardiomyocytes and flow cytometry

Madhubanti Mullick, Dwaipayan Sen
Hypoxia-reperfusion (H/R) emblems a plethora of pathological conditions which is potent in contributing to the adversities encountered by human mesenchymal stem cells (hMSCs) in post-transplant microenvironment, resulting in transplant failure. D-Alanine 2, Leucine 5 Enkephaline (DADLE)-mediated delta opioid receptor (DOR) activation is well-known for its recuperative properties in different cell types like neuronal and cardiomyocytes. In the current study its effectiveness in assuaging hMSC mortality under H/R-like insult has been delineated...
April 12, 2018: Stem Cell Reviews
Yi Shen, Gang Tang, Pan Gao, Bin Zhang, Hang Xiao, Liang-Yi Si
High glucose plays a vital role in apoptosis in H9C2 cells. However, the exact molecular mechanism remains unclear. In this study, we aimed to evaluate the cardio-protective role of A2b receptor in high glucose-induced cardiomyocyte apoptosis via PI3K/Akt pathway. Adenosine A2b receptor agonist (Bay506583), antagonist (MRS1754), and Akt inhibitor (LY294002) were applied respectively to H9C2 cells before exposed to high glucose for 12 h. Apoptosis of H9C2 cells was determined by TUNEL assay and the apoptosis rate by flow cytometry...
April 6, 2018: In Vitro Cellular & Developmental Biology. Animal
Qiang Su, Junjing Yao, Cunjian Sheng
Myocarditis is a cardiomyopathy associated with inflammatory response. It has been reported that geniposide (GEN), a traditional Chinese herb extract from Gardenia jasminoides Ellis, possesses an anti-inflammatory effect and a protective effect on cardiomyocytes. The present study aimed to explore the protective role of GEN and the underlying mechanism in LPS-injured H9c2 cells. H9c2 cells were treated with LPS to induce cell injury and then we investigated the effect of GEN. miR-145 expression was inhibited by transfection with miR-145 inhibitor and its expression was measured by RT-PCR...
April 2, 2018: Inflammation
Zhongli Wang, Rongsheng Cui, Kai Wang
The protective effect of sevoflurane on apoptosis of rat H9c2 cardiomyocytes induced by H2 O2 and the effect on the expression of glucose-regulated protein 78 (GRP78) were investigated. H9c2 cells were routinely cultured and divided into the control, model and sevoflurane groups. Cells in the model group were treated with 400 µM H2 O2 , and cells in the sevoflurane group were pretreated with sevoflurane prior to treatment with 400 µM H2 O2 . MTT assay was used to assess cell viability. Annexin V-propidium iodide (AV-PI) double staining flow cytometry was used to detect apoptosis...
March 2018: Experimental and Therapeutic Medicine
Yong Zhao, Chun-Hua Ding
Background: Recent research indicates that nerve growth factor (NGF) promotes cardiac repair following myocardial infarction by promoting angiogenesis and cardiomyocyte survival. The purpose of this study was to investigate the effects of NGF on cardiac fibroblasts (CFs) proliferation, cell cycle, migration, and myofibroblast transformation in vitro. Methods: CFs were obtained from ventricles of neonatal Sprague-Dawley rats and incubated with various concentrations of NGF (0, 0...
April 5, 2018: Chinese Medical Journal
Thi Y L Le, Masahito Ogawa, Eddy Kizana, Jenny E Gunton, James J H Chong
BACKGROUND: Vitamin D has been implicated in the prevention of heart failure. However the underlying mechanism remains unclear. We hypothesised that these effects may be partially mediated by cardiac stem/progenitor cells (CPCs). Therefore, we examined the effects of 1,25-dihydroxyvitamin D3 (1,25D) on cell cycle activity and differentiation of a previously described CPC population called cardiac colony-forming unit fibroblasts (cCFU-Fs). METHODS: cCFU-Fs were isolated from adult male C57Bl/6 mouse hearts using fluorescence-activated cell sorting...
February 3, 2018: Heart, Lung & Circulation
Xue-Ru Liu, Tao Li, Lu Cao, Yi-Yan Yu, Lin-Lin Chen, Xue-Hui Fan, Bin-Bin Yang, Xiao-Qiu Tan
Dexmedetomidine (DEX), an α2 adrenoceptor agonist, has sedative and analgesic properties and myocardial protective effects. However, the mechanism underlying the protective effects of DEX on the myocardium remain unclear. The present study aimed to determine whether DEX serves an important role on cardioprotection through the endoplasmic reticulum (ER)‑ and mitochondria‑mediated apoptosis signaling pathways. Neonatal rat cardiomyocytes (NRCMs) were cultured and divided four groups: i) Normal culture medium with 10% fetal bovine serum (control group); ii) H2O2 at 500 µM (H2O2 group); iii) DEX at 5 µM (DEX group); and iv) H2O2 plus DEX (H2O2 + DEX group)...
March 15, 2018: Molecular Medicine Reports
Shenjie Chen, Bingsheng Yang, Yifei Xu, Yiqing Rong, Yuangang Qiu
Myocardial hypertrophy is often associated with myocardial infarction. Luteolin-7-O-glucoside (LUTG) has the prosperity of preventing cardiomyocyte injury. The current study aimed to explore the potential protective effect of LUTG and its relevant mechanisms in the heart. To establish the cardiac hypertrophy model in vitro, Angiotensin II (Ang II) was used to stimuli H9c2 cells in this study. The CCK‑8 assay showed that LUTG pretreatment improved cell viability of cardiomyocytes co‑treated with Ang II and ischemia/reperfusion...
March 19, 2018: Molecular Medicine Reports
Akira Yoshida, Jong-Kook Lee, Satoki Tomoyama, Keiko Miwa, Keiichi Shirakawa, Sanae Hamanaka, Tomoyuki Yamaguchi, Hiromitsu Nakauchi, Shigeru Miyagawa, Yoshiki Sawa, Issei Komuro, Yasushi Sakata
Aims: The aim of the present study is to develop in vitro experimental analytical method for the electrophysiological properties of allogeneic induced pluripotent stem cell-derived cardiomyocytes (CMs) in cardiac conduction defect model. Methods and results: Cardiomyocytes were derived from rat induced pluripotent stem cells CMs (riPSC-CMs) using an embryoid body-based differentiation method with the serial application of growth factors including activin-A, bone morphogenetic protein 4 (BMP-4), and inhibitor of wnt production 2 (IWP-2)...
March 15, 2018: Europace: European Pacing, Arrhythmias, and Cardiac Electrophysiology
Wen Hao, Shutian Shi, Shenghui Zhou, Xiao Wang, Shaoping Nie
This study aimed to examine the effects of aspirin on the growth and cardiomyocyte differentiation grade of bone marrow mesenchymal stem cells (BMMSCs). BMMSCs were divided into five differentiation groups with different concentrations of aspirin (0mM, 0.5mM, 1mM, 2mM, or 5mM), and a undifferentiated control group. Cell growth was measured by cell proliferation, apoptosis assays and DNA cycle analysis. The differentiation grade of BMMSC-derived cardiomyocyte-like cells was examined by measuring the levels of cardiac-specific proteins with cyto-immunofluorescence staining, flow cytometry, and Western blotting...
March 15, 2018: European Journal of Pharmacology
Holger Lörchner, Christian Widera, Yunlong Hou, Albrecht Elsässer, Henning Warnecke, Evangelos Giannitsis, Jean-Sebastien Hulot, Thomas Braun, Kai C Wollert, Jochen Pöling
BACKGROUND: Regenerating islet-derived protein 3 beta (Reg3β) is a cardiomyocyte-derived chemokine for macrophages that is upregulated after myocardial infarction (MI) in mice. Here, we hypothesized that monitoring Reg3β expression might provide specific information on the degree of cardiac inflammation, which is a key determinant in disease progression and prognosis of patients with acute coronary syndrome (ACS). METHODS AND RESULTS: The expression of Reg3β and other inflammatory markers including C-reactive protein (CRP) and myeloperoxidase (MPO) was measured by immunoblotting at serial time points in the hearts and serum of mice with acute MI...
May 1, 2018: International Journal of Cardiology
Meiling Yan, Kankai Chen, Li He, Shuai Li, Dong Huang, Jingbo Li
BACKGROUND/AIMS: Hyperuricemia is associated with an increased risk for multiple cardiovascular diseases, but the underlying mechanisms remain largely elusive. Calpain-1 is a protease that is implicated in several pathological conditions that affect the heart. The aim of this current study was to test the effects of uric acid (UA) on cardiomyocyte survival and cardiac function and to investigate the role of calpain-1 in the UA-induced effects in the heart and their underlying mechanisms...
March 7, 2018: Cellular Physiology and Biochemistry
Nagako Sougawa, Shigeru Miyagawa, Satsuki Fukushima, Ai Kawamura, Junya Yokoyama, Emiko Ito, Akima Harada, Kaori Okimoto, Noriko Mochizuki-Oda, Atsuhiro Saito, Yoshiki Sawa
Induced pluripotent stem cells (iPSCs) are promising candidate cells for cardiomyogenesis in the failing heart. However, teratoma/tumour formation originating from undifferentiated iPSCs contaminating the graft is a critical concern for clinical application. Here, we hypothesized that brentuximab vedotin, which targets CD30, induces apoptosis in tumourigenic cells, thus increasing the safety of iPSC therapy for heart failure. Flow cytometry analysis identified consistent expression of CD30 in undifferentiated human iPSCs...
February 27, 2018: Scientific Reports
Qiuling Xiang, Yan Liao, Hua Chao, Weijun Huang, Jia Liu, Haixuan Chen, Dongxi Hong, Zhengwei Zou, Andy Peng Xiang, Weiqiang Li
BACKGROUND: The LIM-homeobox transcription factor islet-1 (ISL1) has been proposed as a marker for cardiovascular progenitor cells. This study investigated whether forced expression of ISL1 in human mesenchymal stem cells (hMSCs) improves myocardial infarction (MI) treatment outcomes. METHODS: The lentiviral vector containing the human elongation factor 1α promoter, which drives the expression of ISL1 (EF1α-ISL1), was constructed using the Multisite Gateway System and used to transduce hMSCs...
February 26, 2018: Stem Cell Research & Therapy
Mirnalini Sharma, Baijayantimala Mishra, Uma Nahar Saikia, Ajay Bahl, R K Ratho
Background & objectives: Coxsackievirus B (CVB), a member of human Enterovirus group, is the most common cause of viral myocarditis. Coxsackievirus adenovirus receptor (CAR) is identified as a key determinant for the entry of CVB in the target cells. Thus, blockade of receptor by RNA interference (RNAi) may inhibit the entry and pathogenesis of CVB in cardiac cells. The present study was aimed to determine the effect of CAR small dsRNA (siRNA) on coxsackieviral load and CAR expression in coxsackievirus-infected cardiomyocytes...
October 2017: Indian Journal of Medical Research
Venkata Naga Srikanth Garikipati, Saurabh Pratap Singh, Yamuna Mohanram, Ashwani Kumar Gupta, Deepa Kapoor, Soniya Nityanand
BACKGROUND: Mesenchymal stem cells (MSCs) are promising cells for cardiovascular regenerative medicine. However, their potential may be limited, because of their restricted cardiovascular differentiation potential and decline in their number and functional characteristics with increasing donor age. We have previously shown that rat fetus heart harbors primitive MSCs and administration of these cells improved left ventricular (LV) function after ischemia/reperfusion injury in rats. To evaluate their potential as a new cell type for clinical cardiovascular cell therapy, we have undertaken this study on the isolation and characterization of human fetal cardiac MSCs (hfC-MSCs)...
2018: PloS One
Pengfei Li, Zhuyuan Liu, Yuan Xie, Huanyu Gu, Qiying Dai, Jianhua Yao, Lei Zhou
Exosomes are small-sized vesicles that can be released from cells into the serum. Exosomes play important roles in regulating many biological processes including cell proliferation, apoptosis, cell cycle, and metabolism. However, the roles and mechanisms of plasma exosomes in the apoptosis of rat H9C2 cardiomyocytes are largely unknown. In this study, we isolated plasma exosomes as confirmed by the marker protein CD63. Using flow cytometry and western blot analysis, we found that exosomes attenuated hydrogen peroxide (H2O2)-induced apoptosis and improved survival of rat H9C2 cardiomyocytes...
February 5, 2018: Journal of Cardiovascular Translational Research
Mingxia Gu, Jing Wang, Yi Wang, Yanjuan Xu, Yingqiang Zhang, Weiqing Wu, Shuping Liao
Recently, microRNAs (miRNAs) have been shown to involve in the process of heart failure. This study aims to investigate the functional role of miR-147b in rat H9c2 cardiomyocytes and explore the underlying molecular mechanisms. Cell viability of H9c2 cells was detected by MTT assay. Cell apoptosis was detected by flow cytometry. Expression of miR-147b and KLF13 mRNA was detected by quantitative real-time PCR. The relationship between miR-147b and KLF13 was verified by dual-luciferase reporter assay. Protein levels were detected by western blot analysis...
January 25, 2018: Acta Biochimica et Biophysica Sinica
Yuqiao Chang, Cixia Li, Yangyang Jia, Peng Chen, Yonglong Guo, Airong Li, Zhikun Guo
OBJECTIVE: To explore the differentiation tendency of CD90+ cardiac fibroblast (CFs) into cardiomyogenic cells in vitro and repair functions in acute myocardial infarction rats. METHODS: CD90+ subpopulation was sorted from rat CFs by flow cytometry. 10 μmoL/L of 5-Azacytosine (5-aza) was used to induce differentiation of CFs into cardiomyogenic cells. An acute myocardial infarction model was prepared by ligation of the rat left anterior descending coronary artery...
March 2018: International Journal of Biochemistry & Cell Biology
Xiuge Gao, Yani Zheng, Xiangchun Ruan, Hui Ji, Lin Peng, Dawei Guo, Shanxiang Jiang
Salinomycin, as a polyether ionophore antibiotic, is extensively used as a feed additive against coccidiosis in poultry and as a growth promoter of ruminants worldwide. Owing to its narrow therapeutic index, numerous intoxication have been reported in target/non-target animals by overdosage, misuse or drug interactions as well as human who consumed salinomycin accidently. Salinomycin-induced cardiotoxicity in chicken and non-target animals is considered as a major contributor to animal death. In the current study, we aim to elucidate the underlying mechanism of its myocardial toxicity using primary chicken myocardial cell as an in vitro model...
February 25, 2018: Chemico-biological Interactions
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