scholarly journals Palm Tocotrienol-Rich Fraction Protects Neonatal Rat Cardiomyocytes from H2o2-Induced Oxidative Damage

2021 ◽  
Vol 50 (5) ◽  
pp. 1457-1466
Author(s):  
Noor Shareena Aisha Abdul Khalid ◽  
Khuzaidatul Azidah Ahmad Nazri ◽  
Zakiah Jubri
Keyword(s):  
Cell Death ◽  
Heart Disease ◽  
Oxidative Damage ◽  
Neonatal Rat ◽  
Green Fluorescence ◽  
Sod Activity ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes ◽  
Pre Treatment ◽  
Tocotrienol Rich Fraction

Oxidative stress plays an important role in the pathogenesis of heart disease. Tocotrienol-rich fraction (TRF) is an antioxidant and that has the potential to reduce the risk of heart disease. This study is to determine the protective effects of palm TRF against H2O2-induced oxidative damage in neonatal rat cardiomyocytes (NRCM). The NRCM were divided into control, treated with TRF (10 µg/mL), H2O2 (0.5 mM) and treated with TRF prior to H2O2 induction (pre-treatment). Cell viability was determined by the MTS assay,while the presence of reactive oxygen species (ROS) was determined using fluorescent dihydroethidium (DHE) and 5-(and-6)-carboxy-2′,7′-dichlorodihydrofluorescein diacetate (carboxy-H2DCFDA) dye. Mitochondrial integrity and cell death were determined using JC-1 and Annexin V-FITC staining, respectively. Lactate dehydrogenase (LDH) and superoxide dismutase (SOD) activity were determined by colorimetric assay kit. The concentration of H2O2 from 0.5 to 5 mM reduced the cell viability and the H2O2 IC50 value of 0.5 mM was used in the experiment. H2O2 induction increased the intensity of carboxy-H2DCFDA and DHE-stains; and also the intensity of green fluorescence of J-monomers in JC-1 staining compared to the control group. The activity of LDH increased while the activity of SOD decreased in the H2O2 group. Pre-treatment with TRF reduced the intensities of carboxy-H2DCFDA and DHE-stains, as well as the green fluorescence of J-monomers in JC-1. Meanwhile, the LDH activity was reduced in the pre-treatment group but no changes were recorded in SOD activity compared to the H2O2 group. Palm TRF protects cardiomyocytes from oxidative damage by reducing ROS production and maintaining the mitochondrial membrane integrity thus reducing cell death.

Role of Calcium-Activated Neutral Protease (Calpain) in Cell Death in Cultured Neonatal Rat Cardiomyocytes During Metabolic Inhibition

1995 ◽  
Vol 76 (6) ◽  
pp. 1071-1078 ◽  
Author(s):  
Douwe E. Atsma ◽  
E.M. Lars Bastiaanse ◽  
Anastazia Jerzewski ◽  
Lizet J.M. Van der Valk ◽  
Arnoud Van der Laarse
Keyword(s):  
Cell Death ◽  
Metabolic Inhibition ◽  
Neonatal Rat ◽  
Neutral Protease ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes

Low external pH limits cell death of energy-depleted cardiomyocytes by attenuation of Ca2+ overload

1996 ◽  
Vol 270 (6) ◽  
pp. H2149-H2156 ◽  
Author(s):  
D. E. Atsma ◽  
E. M. Bastiaanse ◽  
L. Van der Valk ◽  
A. Van der Laarse
Keyword(s):  
Cell Death ◽  
Cell Injury ◽  
Metabolic Inhibition ◽  
Intracellular Concentration ◽  
Neonatal Rat ◽  
Atp Content ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes ◽  
External Ph

We studied the effect of external pH (pHe) on cell injury, ATP content, and intracellular concentration of Ca2+ ([Ca2+]i), Na+ ([Na+]i), and H+ (pHi) during metabolic inhibition (NaCN + 2-deoxyglucose) in neonatal rat cardiomyocytes. Cell death during metabolic inhibition decreased at pHe < 7.4, with almost no cell death at pHe 6.0. Lowering pHe resulted in only temporary ATP conservation. During metabolic inhibition at pHe 7.4, [Ca2+]i rose from 86 +/- 44 nM to 2.5 +/- 0.4 microM, but at pHe 6.0, [Ca2+]i rose to only 510 +/- 215 nM. During metabolic inhibition at pHe 7.4, pHi decreased from 7.25 +/- 0.06 to 6.82 +/- 0.16, but at pHe 6.0, pHi decreased to 6.34 +/- 0.17. During metabolic inhibition at pHe 7.4, [Na+]i increased from 9.1 +/- 0.86 to 26.1 +/- 4.1 mM. At pHe 6.0, [Na+]i rose more rapidly, to 27.3 +/- 3.5 mM. At pHe < 7.4, sarcolemmal Na+/Ca2+ exchanger activity, involved in the development of Ca2+ overload, was decreased, as assessed during Na(+)-free incubation. We conclude that low pHe protects cardiomyocytes during metabolic inhibition by limiting Ca2+ overload via Na+/Ca2+ exchanger inhibition.

Imatinib Induces Autophagy Mediated Cell Death in Neonatal Rat Cardiomyocytes

2009 ◽  
Vol 15 (7) ◽  
pp. S160-S161
Author(s):  
Yugo Uchida ◽  
Miyuki Kobara ◽  
Syouta Tsujimoto ◽  
Kenta Tanaka ◽  
Hiroe Toba ◽  
...  
Keyword(s):  
Cell Death ◽  
Neonatal Rat ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes

scholarly journals The protective effect of palm tocotrienol-rich fraction against H2O2- induced oxidative stress in neonatal rat cardiomyocytes

2017 ◽  
Author(s):  
Noor Shareena Aisha Abdul Khalid ◽  
Zakiah Jubri
Keyword(s):  
Oxidative Stress ◽  
Membrane Potential ◽  
Mitochondrial Membrane ◽  
Neonatal Rat ◽  
Ros Production ◽  
Rat Cardiomyocytes ◽  
Apoptotic Gene ◽  
Neonatal Rat Cardiomyocytes ◽  
Ldh Activity ◽  
Tocotrienol Rich Fraction

Background: Oxidative stress plays an important role in the pathogenesis of heart diseases. Antioxidants such as palm tocotrienol-rich fraction (TRF) can reduce oxidative stress, hence preventing and reducing the risk of heart disease. This study was aimed to determine the protective effects of TRF against hydrogen peroxide (H2O2 ) - induced oxidative stress in neonatal rat cardiomyocytes (NRCM). Methods: The NRCM were divided into five groups: (1) control, (2) cells treated with TRF (10 µg/ml) for 24 hours, (3) cells subjected to H2O2 (0.5 mM ) for 30 minutes, (4) cells pre-treated with TRF, and (5) cells post-treated with TRF. The IC50 of H2O2 (0 – 5 mM ) and the effective dose of TRF (0 – 25 µg/ml) were determined using the MTS cell viability assay. Meanwhile, ELISA was used to measure the level of reactive oxygen species (ROS). The presence of superoxides and H2O2 were detected by dihydroethidium and 5-(and-6 ) - carboxy -2′,7′-dichlorodihydrofluorescein diacetate respectively. Flowcytometry analysis was conducted to determine the presence of apoptosis and measure the mitochondrial membrane potential, whereby the former involved the use of Annexin V-FITC stain while the latter JC-1 stain. The gene expressions of antioxidant (SOD, CAT, GPx) and apoptosis (Bax, Bcl-2, Caspase-3) enzymes were studied using qRT -PCR. Results: The IC50 of H2O2 was 0.5 mM while the effective dose of TRF 10 µg/ml. The cells which were subjected to H2O2 showed a decrease in NRCM viability and significant increase (p < 0.05) in ROS production. LDH activity and green fluorescence intensity (which indicated mitochondrial depolarisation ) were increased following H2O2 induction . With reference to the control, the H2O2- induced group had a higher percentage of late apoptotic cells, which was associated with the upregulation of the pro-apoptotic gene, Bax, and downregulation of the anti-apoptotic gene, Bcl-2 (p < 0.05). H2O2 also upregulated GPx expression , apart from downregulating CAT and Cu/Zn SOD expression (p < 0.05). The pre- and post-treatment groups had increased cell viability and reduced ROS production. Pre-treatment with TRF protected the cell membranes and mitochondria from H2O2- induced injury, as reflected by the reduction in extracellular LDH activity and apoptosis (the latter of which was associated with the downregulation of Bax). Meanwhile, the expression of GPx, Cat, and Cu/Zn SOD was reduced in the post-treatment group. Conclusion: By scavenging for ROS, palm TRF directly protects the cell membrane from H2O2- induced injury, leading to a decrease in oxidative stress. Thus, palm TRF maintains the mitochondrial membrane potential and prevents apoptosis secondary to decreased Bax expression.

scholarly journals The protective effect of palm tocotrienol-rich fraction against H2O2- induced oxidative stress in neonatal rat cardiomyocytes

2017 ◽  
Author(s):  
Noor Shareena Aisha Abdul Khalid ◽  
Zakiah Jubri
Keyword(s):  
Oxidative Stress ◽  
Membrane Potential ◽  
Mitochondrial Membrane ◽  
Neonatal Rat ◽  
Ros Production ◽  
Rat Cardiomyocytes ◽  
Apoptotic Gene ◽  
Neonatal Rat Cardiomyocytes ◽  
Ldh Activity ◽  
Tocotrienol Rich Fraction

Background: Oxidative stress plays an important role in the pathogenesis of heart diseases. Antioxidants such as palm tocotrienol-rich fraction (TRF) can reduce oxidative stress, hence preventing and reducing the risk of heart disease. This study was aimed to determine the protective effects of TRF against hydrogen peroxide (H2O2 ) - induced oxidative stress in neonatal rat cardiomyocytes (NRCM). Methods: The NRCM were divided into five groups: (1) control, (2) cells treated with TRF (10 µg/ml) for 24 hours, (3) cells subjected to H2O2 (0.5 mM ) for 30 minutes, (4) cells pre-treated with TRF, and (5) cells post-treated with TRF. The IC50 of H2O2 (0 – 5 mM ) and the effective dose of TRF (0 – 25 µg/ml) were determined using the MTS cell viability assay. Meanwhile, ELISA was used to measure the level of reactive oxygen species (ROS). The presence of superoxides and H2O2 were detected by dihydroethidium and 5-(and-6 ) - carboxy -2′,7′-dichlorodihydrofluorescein diacetate respectively. Flowcytometry analysis was conducted to determine the presence of apoptosis and measure the mitochondrial membrane potential, whereby the former involved the use of Annexin V-FITC stain while the latter JC-1 stain. The gene expressions of antioxidant (SOD, CAT, GPx) and apoptosis (Bax, Bcl-2, Caspase-3) enzymes were studied using qRT -PCR. Results: The IC50 of H2O2 was 0.5 mM while the effective dose of TRF 10 µg/ml. The cells which were subjected to H2O2 showed a decrease in NRCM viability and significant increase (p < 0.05) in ROS production. LDH activity and green fluorescence intensity (which indicated mitochondrial depolarisation ) were increased following H2O2 induction . With reference to the control, the H2O2- induced group had a higher percentage of late apoptotic cells, which was associated with the upregulation of the pro-apoptotic gene, Bax, and downregulation of the anti-apoptotic gene, Bcl-2 (p < 0.05). H2O2 also upregulated GPx expression , apart from downregulating CAT and Cu/Zn SOD expression (p < 0.05). The pre- and post-treatment groups had increased cell viability and reduced ROS production. Pre-treatment with TRF protected the cell membranes and mitochondria from H2O2- induced injury, as reflected by the reduction in extracellular LDH activity and apoptosis (the latter of which was associated with the downregulation of Bax). Meanwhile, the expression of GPx, Cat, and Cu/Zn SOD was reduced in the post-treatment group. Conclusion: By scavenging for ROS, palm TRF directly protects the cell membrane from H2O2- induced injury, leading to a decrease in oxidative stress. Thus, palm TRF maintains the mitochondrial membrane potential and prevents apoptosis secondary to decreased Bax expression.

Azelnidipine Inhibits H2O2-Induced Cell Death in Neonatal Rat Cardiomyocytes

2007 ◽  
Vol 21 (1) ◽  
pp. 69-72 ◽  
Author(s):  
Yo Koyama ◽  
Yasuchika Takeishi ◽  
Hiroki Takahashi ◽  
Tetsuro Shishido ◽  
Takanori Arimoto ◽  
...  
Keyword(s):  
Cell Death ◽  
Neonatal Rat ◽  
Rat Cardiomyocytes ◽  
Neonatal Rat Cardiomyocytes

scholarly journals Effects of endothelin-11-31 on cell viability and [Ca2+]i in cultured neonatal rat cardiomyocytes

2008 ◽  
pp. 373-378
Author(s):  
A-J Ren ◽  
X Yuan ◽  
L Lin ◽  
Y-X Pan ◽  
Y-W Qing ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cell Injury ◽  
Neonatal Rat ◽  
Sod Activity ◽  
Rat Cardiomyocytes ◽  
Continuous Increase ◽  
Neonatal Rat Cardiomyocytes ◽  
Eta Receptor ◽  
Laser Confocal Microscope ◽  
Eta Receptor Antagonist

We previously found that Endothelin-11-31 (ET-11-31) exhibited a pro-arrhythmogenic effect in isolated rat hearts. In this study, we further investigated the effects of ET-11-31 on a cell viability and observed [Ca2+]i in cultured cardiomyocytes. Cultured neonatal rat cardiomyocytes were treated with 0.1, 1, and 10 nM ET-11-31 for 24h in the presence or absence of ETA receptor antagonist (BQ123) or phosphoramidon, a NEP/ECE inhibitor. Cell injury was evaluated by supernatant lactate dehydrogenase (LDH) assay, superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content. Cell viability was assessed by MTT assay. [Ca2+]i was measured with Fluo-3/AM under a laser confocal microscope. 1) ET-11-31 dose-dependently increased LDH release and decreased cell viability. 2) LDH and MDA levels were significantly elevated and SOD activity decreased after administration of 1 nM ET-11-31 for 24h, and these changes were markedly attenuated by 1 uM BQ123. 3) Exposure to 10 nM ET-11-31 caused a continuous increase in [Ca2+]i to cultured beating cardiomyocytes and termination of [Ca2+]i transient within 6 min, and this change was reversed by 1 uM BQ123 and attenuated by 0.5 mM phosphoramidon. These results suggest that ET-11-31 could cause cell injury, and that the effect of ET-11-31 on [Ca2+]i transients is mainly mediated by ETA receptor and partially attributed to the conversion of ET-11-31 to ET-11-21.

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