• Title/Summary/Keyword: Mitochondrial stress

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Neuropeptide Y protects kidney against cisplatin-induced nephrotoxicity by regulating p53-dependent apoptosis pathway

  • Kim, Namoh;Min, Woo-Kie;Park, Min Hee;Lee, Jong Kil;Jin, Hee Kyung;Bae, Jae-sung
    • BMB Reports
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    • v.49 no.5
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    • pp.288-292
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    • 2016
  • Cisplatin is a platinum-based chemotherapeutic drug for treating various types of cancers. However, the use of cisplatin is limited by its negative effect on normal tissues, particularly nephrotoxicity. Various mechanisms such as DNA adduct formation, mitochondrial dysfunction, oxidative stress, and apoptosis are involved in the adverse effect induced by cisplatin treatment. Several studies have suggested that neuropeptide Y (NPY) is involved in neuroprotection as well as restoration of bone marrow dysfunction from chemotherapy induced nerve injury. However, the role of NPY in chemotherapy-induced nephrotoxicity has not been studied. Here, we show that NPY rescues renal dysfunction by reducing the expression of pro-apoptotic proteins in cisplatin induced nephrotoxicity through Y1 receptor, suggesting that NPY can protect kidney against cisplatin nephrotoxicity as a possible useful agent to prevent and treat cisplatin-induced nephrotoxicity.

Protective Effects of Natural Phytochemicals on the Lipid Peroxides Induced Apoptosis in the Human Endothelial ECV 304 Cells

  • Kim, Ae-Jung;Kim, Mae-Wha;Kang, Young-Hee;Lee, Myoung-Sook
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.436-441
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    • 2009
  • The final bio-metabolites of lipid peroxidation (LPO) such as 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA) have been suggested to mediate the oxidative stress-linked pathological incidences. Natural phytochemicals such as polyphenolic compounds in green tea have been known in preventing the LPO induced cellular growth inhibition and apoptosis. We investigated that green tea ethanol extracts (GTE) inhibit LPO-induced apoptosis in ECV 304 cells. GTE had time- or dose-dependent anti-apoptotic effects as evidenced by changes in cell morphology, MTT assay, DNA fragmentation, LPO production, and the Western blotting for apoptotic expression. In the 4-HNE-induced apoptosis model, GTE $10-20{\mu}g/mL$ decreased cell death through decreasing LPO production. GTE protected 4-HNE induced apoptosis, as evidence with down regulation of mitochondrial signaling such as cytochrome C and caspase-3 activity. GTE increased bcl2, survival signaling protein, compared to 4-HNE alone within 6 hr incubation. Since polyphenols in GTE are effective antioxidants in endothelial ECV 304 cells, we suggested that natural polyphenols might be anti-atherosclerotic.

Hydrogen Peroxide Induces Apoptosis of BJAB Cells Due to Formation of Hydroxyl Radicals Via Intracellular Iron-mediated Fenton Chemistry in Glucose Oxidase-mediated Oxidative Stress

  • Lee, Jeong-Chae;Son, Young-Ok;Choi, Ki-Choon;Jang, Yong-Suk
    • Molecules and Cells
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    • v.22 no.1
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    • pp.21-29
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    • 2006
  • The aim of this study was to determine if hydrogen peroxide ($H_2O_2$) generated by glucose oxidase (GO) induces apoptosis or necrosis of BJAB cells and which radical is the direct mediator of cell death. We found that GO produced $H_2O_2$ continuously in low concentrations, similar to in vivo conditions, and decreased proliferation and cell viability in a dose-dependent manner. The GO-mediated cytotoxicity resulted from apoptosis, and was confirmed by monitoring the cells after H33342/Annexin V/propidium iodide staining. Decreases of mitochondrial membrane potential and intracellular glutathione level were found to be critical events in the $H_2O_2$-mediated apoptosis. Additional experiments revealed that $H_2O_2$ exerted its apoptotic action through the formation of hydroxyl radicals via the Fenton rather than the Haber-Weiss reaction. Moreover, intracellular redox-active iron, but not copper, participated in the $H_2O_2$-mediated apoptosis.

Isolation and Characterization of Malate Dehydrogenase Gene from Panax ginseng C.A. Meyer (고려인삼에서 Malate Dehydrogenase 유전자의 분리 및 분석)

  • Kim, Yu-Jin;Shim, Ju-Sun;Lee, Jung-Hye;Jung, Dae-Young;In, Jun-Gyo;Lee, Bum-Soo;Min, Byung-Hoon;Yang, Deok-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.4
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    • pp.261-267
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    • 2008
  • Malate dehydrogenase is a ubiquitous enzyme in plants, involving in a range of metabolic processes depending on its subcellular location. A malate dehydrogenase (PgMDH) cDNA was isolated and characterized from the root of Panax ginseng C. A. Meyer. The deduced amino acid sequence of PgMDH showed high similarity with the NAD-dependent mitochondrial malate dehydrogenase from Glycinemax (P17783), Eucalyptus gunnii (P46487), and Lycopersicon esculentum (AAU29198). And the segment of a malate dehydrogenase gene was amplified through RT-PCR. The expression of PgMDH was increased after treatments of chilling, salt, UV, cadmium or copper treatment.

Water Extract of Ash Tree (Fraxinus rhynchophylla) Leaves Protects against Paracetamol-Induced Oxidative Damages in Mice

  • Jeon, Jeong-Ryae
    • Food Science and Biotechnology
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    • v.15 no.4
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    • pp.612-616
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    • 2006
  • The protective effect of water extract of ash tree leaves (ALE) against oxidative damages was investigated in paracetamol-induced BALB/c mice. Biochemical analysis of anti-oxidative enzymes, immunoblot analyses of hepatic cytochrome P450 2El (CYP2E1), and the gene expression of tumor necrosis factor (TNF-${\alpha}$) were examined to determine the extract's protective effect and its possible mechanisms. BALB/c mice were divided into three groups: normal, paracetamol-administered, and ALE-pretreated groups. A single dose of paracetamol led to a marked increase in lipid peroxidation as measured by malondialdehyde (MDA). This was associated with a significant reduction in the hepatic antioxidant system, e.g., glutathione (GSH). Paracetamol administration also significantly elevated the expression of CYP2E1, according to immunoblot analysis, and of TNF-${\alpha}$ mRNA in liver. However, ALE pretreatment prior to the administration of paracetamol significantly decreased hepatic MDA levels. ALE restored hepatic glutathione and catalase levels and suppressed the expression of CYP2E1 and TNF-${\alpha}$ observed in inflammatory tissues. Moreover, ALE restored mitochondrial ATP content depleted by the drug administration. These results show that the extract of ash tree leaves protects against paracetamol-induced oxidative damages by blocking oxidative stress and CYP2E1-mediated paracetamol bioactivation.

Effect of Bojungikgi-tang on cytochrome P450 and LKB1-AMPK anti-oxidant signaling pathway (보중익기탕이 cytochrome P450 및 LKB1-AMPK 항산화 신호에 미치는 영향)

  • Song, Yu Rim;Park, Sun-Dong;Kim, Young Woo
    • Herbal Formula Science
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    • v.29 no.4
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    • pp.277-283
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    • 2021
  • Objectives : We investigated the effects of Bojungikgi-tang (BJIGT) on P450 cytochrome enzyme and oxidative stress in the cells. Methods : We enrolled the HepG2 hepatocyte cell line to assess MTT assay, flow cytometer, and immunoblotting analysis. Expression of CYP450 was confirmed by immunoblotting analysis in the Huh7 cell line. Results : We determined that BJIKT markdely changed the expression of the CYP2C19, CYP2D6, and CYP2E1. Moreover, BJIKT inhibited the cell toxicity induced by arachidonic acid + iron treatment, as assessed by FACS analysis. BJIKT induced AMPK activation, which increased the phophorylation of ACC. Conclusions : This study verified the effects of BJIKT, on P450, ROS production, mitochondrial damage and AMPK signaling pathway, which might give us the scientific information about the traditional herbal prescription.

N-terminal formylmethionine as a novel initiator and N-degron of eukaryotic proteins

  • Kim, Jeong-Mok
    • BMB Reports
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    • v.52 no.3
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    • pp.163-164
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    • 2019
  • The ribosomal synthesis of proteins in the eukaryotic cytosol has always been thought to start from the unformylated N-terminal (Nt) methionine (Met). In contrast, in virtually all nascent proteins in bacteria and eukaryotic organelles, such as mitochondria and chloroplasts, Nt-formyl-methionine (fMet) is the first building block of ribosomal synthesis. Through extensive approaches, including mass spectrometric analyses of the N-termini of proteins and molecular genetic techniques with an affinity-purified antibody for Nt-formylation, we investigated whether Nt-formylated proteins could also be produced and have their own metabolic fate in the cytosol of a eukaryote, such as yeast Saccharomyces cerevisiae. We discovered that Nt-formylated proteins could be generated in the cytosol by yeast mitochondrial formyltransferase (Fmt1). These Nt-formylated proteins were massively upregulated in the stationary phase or upon starvation for specific amino acids and were crucial for the adaptation to specific stresses. The stress-activated kinase Gcn2 was strictly required for the upregulation of Nt-formylated proteins by regulating the activity of Fmt1 and its retention in the cytosol. We also found that the Nt-fMet residues of Nt-formylated proteins could be distinct N-terminal degradation signals, termed fMet/N-degrons, and that Psh1 E3 ubiquitin ligase mediated the selective destruction of Nt-formylated proteins as the recognition component of a novel eukaryotic fMet/N-end rule pathway, termed fMet/N-recognin.

Functions of PUF Family RNA-Binding Proteins in Aspergillus nidulans

  • Son, Sung-Hun;Jang, Seo-Yeong;Park, Hee-Soo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.676-685
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    • 2021
  • RNA-binding proteins are involved in RNA metabolism and posttranscriptional regulation of various fundamental biological processes. The PUF family of RNA-binding proteins is highly conserved in eukaryotes, and its members regulate gene expression, mitochondrial biogenesis, and RNA processing. However, their biological functions in Aspergillus species remain mostly unknown in filamentous fungi. Here we have characterized the puf genes in the model organism Aspergillus nidulans. We generated deletion mutant strains for the five putative puf genes present in the A. nidulans genome and investigated their developmental phenotypes. Deletion of pufA or pufE affected fungal growth and asexual development. pufA mutants exhibited decreased production of asexual spores and reduced mRNA expression of genes regulating asexual development. The pufE deletion reduced colony growth, increased formation of asexual spores, and delayed production of sexual fruiting bodies. In addition, the absence of pufE reduced both sterigmatocystin production and the mRNA levels of genes in the sterigmatocystin cluster. Finally, pufE deletion mutants showed reduced trehalose production and lower resistance to thermal stress. Overall, these results demonstrate that PufA and PufE play roles in the development and sterigmatocystin metabolism in A. nidulans.

Application of the modified handmade cloning technique to pigs

  • Lee, Eun Ji;Ji, Kuk Bin;Lee, Ji Hye;Oh, Hyun Ju;Kil, Tae Young;Kim, Min Kyu
    • Journal of Animal Science and Technology
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    • v.63 no.2
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    • pp.281-294
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    • 2021
  • Although somatic cell nuclear transfer (SCNT) is frequently employed to produce cloned animals in laboratories, this technique is expensive and inefficient. Therefore, the handmade cloning (HMC) technique has been suggested to simplify and advance the cloning process, however, HMC wastes many oocytes and leads to mitochondrial heteroplasmy. To solve these problems, we propose a modified handmade cloning (mHMC) technique that uses simple laboratory equipment, i.e., a Pasteur pipette and an alcohol lamp, applying it to porcine embryo cloning. To validate the application of mHMC to pig cloning, embryos produced through SCNT and mHMC are compared using multiple methods, such as enucleation efficiency, oxidative stress, embryo developmental competence, and gene expression. The results show no significant differences between techniques except in the enucleation efficiency. The 8-cell and 16-cell embryo developmental competence and Oct4 expression levels exhibit significant differences. However, the blastocyst rate is not significantly different between mHMC and SCNT. This study verifies that cloned embryos derived from the two techniques exhibit similar generation and developmental competence. Thus, we suggest that mHMC could replace SCNT for simpler and cheaper porcine cloning.

Effect of aqueous Nigella sativa extract on the functional parameters of post-thaw human spermatozoa during vitrification

  • Nasiri, Zohreh;Ghorbani, Fatemeh;Seify, Mohammad;Sharbati, Aysan
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.2
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    • pp.110-116
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    • 2022
  • Objective: Sperm vitrification leads to the production of reactive oxygen species (ROS) that can damage the functional parameters of sperm. The present study aimed to investigate the antioxidant effect of Nigella sativa extract on motility, plasma membrane function, mitochondrial membrane potential (MMP), DNA damage, and intracellular ROS production. Methods: A total of 20 sperm samples were used. Samples were divided into six experimental groups, including groups with aqueous extract from N. sativa seeds at concentrations of 1% to 6%, a cryopreserved control group, and a fresh control group. Results: Statistical analysis showed significantly higher total sperm motility at concentrations of 3% to 6% than in the vitrified semen control group. Additionally, progressive motility and all motion characteristics at all concentrations were significantly higher than in the vitrified semen control group. The presence of N. sativa seed extract also improved the quality of the sperm parameters assayed in all experimental groups (1%-6%; intracellular ROS production, DNA damage, MMP, and sperm membrane function) compared to the control group. Conclusion: Higher concentrations of N. sativa led to improvements in all sperm parameters and sperm quality. These findings indicate that N. sativa seed extract is effective for improving the quality of sperm after vitrification.