scholarly journals Viscum albumL. Extracts Protects HeLa Cells against Nuclear and Mitochondrial DNA Damage

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Evren Önay-Uçar ◽  
Özlem Erol ◽  
Başak Kandemir ◽  
Elif Mertoğlu ◽  
Ali Karagöz ◽  
...  
Keyword(s):  
Dna Damage ◽  
Mitochondrial Dna ◽  
Hela Cells ◽  
Oxidative Dna Damage ◽  
Nuclear Dna ◽  
Viscum Album ◽  
Mitochondrial Dna Damage ◽  
Methanolic Extracts ◽  
Host Trees ◽  
Lime Tree

Viscum albumL. is a semiparasitic plant grown on trees and widely used for the treatment of many diseases in traditional and complementary therapy. It is well known that some activities ofViscum albumextracts are varied depending on the host trees, such as antioxidant, apoptosis-inducing, anticancer activities of the plant. The aim of the present study is to examine the comparative effects of methanolic extracts ofV. albumgrown on three different host trees (locust tree, lime tree, and hedge maple tree) on H2O2-induced DNA damage in HeLa cells. Oxidative damage in mitochondrial DNA and two nuclear regions was assessed by QPCR assay. The cells were pretreated with methanolic extracts (10 μg/mL) for 48 h, followed by the treatment with 750 μM H2O2for 1 hour. DNA damage was significantly induced by H2O2while it was inhibited byV. albumextracts. All extracts completely protected against nuclear DNA damage. While the extract from lime tree or white locust tree entirely inhibited mitochondrial DNA damage, that from hedge maple tree inhibited by only 50%. These results suggest that methanolic extracts ofV. albumcan prevent oxidative DNA damage, and the activity is dependent on the host tree.

scholarly journals Simultaneous Quantification of Mitochondrial DNA Damage and Copy Number in Circulating Blood: A Sensitive Approach to Systemic Oxidative Stress

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Sam W. Chan ◽  
Simone Chevalier ◽  
Armen Aprikian ◽  
Junjian Z. Chen
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Mitochondrial Dna ◽  
Copy Number ◽  
Structural Damage ◽  
Oxidative Dna Damage ◽  
Nuclear Dna ◽  
Blood Analysis ◽  
Systemic Oxidative Stress ◽  
Wide Range

Systemic oxidative stress is associated with a wide range of pathological conditions. Oxidative DNA damage is frequently measured in circulating lymphocytes. Mitochondrial DNA (mtDNA) is known to be more sensitive to oxidative damage than nuclear DNA but is rarely used for direct measurement of DNA damage in clinical studies. Based on the supercoiling-sensitive real-time PCR method, we propose a new approach for the noninvasive monitoring of systemic oxidative stress by quantifying the mtDNA structural damage and copy number change in isolated lymphocytes in a single test. We show that lymphocytes have significantly less mtDNA content and relatively lower baseline levels of damage than cancer cell lines. In anex vivochallenge experiment, we demonstrate, for the first time, that exogenous H2O2induces a significant increase in mtDNA damage in lymphocytes from healthy individuals, but no repair activity is observed after 1 h recovery. We further demonstrate that whole blood may serve as a convenient alternative to the isolated lymphocytes in mtDNA analysis. Thus, the blood analysis with the multiple mtDNA end-points proposed in the current study may provide a simple and sensitive test to interrogate the nature and extent of systemic oxidative stress for a broad spectrum of clinical investigations.

scholarly journals PCR‐Based Analysis of Mitochondrial DNA Copy Number, Mitochondrial DNA Damage, and Nuclear DNA Damage

2016 ◽  
Vol 67 (1) ◽  
Author(s):  
Claudia P. Gonzalez‐Hunt ◽  
John P. Rooney ◽  
Ian T. Ryde ◽  
Charumathi Anbalagan ◽  
Rashmi Joglekar ◽  
...  
Keyword(s):  
Dna Damage ◽  
Mitochondrial Dna ◽  
Copy Number ◽  
Nuclear Dna ◽  
Dna Copy Number ◽  
Mitochondrial Dna Damage ◽  
Mitochondrial Dna Copy Number

scholarly journals Sepsis is associated with mitochondrial DNA damage and a reduced mitochondrial mass in the kidney of patients with sepsis-AKI

Critical Care ◽  
2021 ◽  
Vol 25 (1) ◽  
Author(s):  
Elisabeth C. van der Slikke ◽  
Bastiaan S. Star ◽  
Matijs van Meurs ◽  
Robert H. Henning ◽  
Jill Moser ◽  
...  
Keyword(s):  
Dna Damage ◽  
Mitochondrial Dna ◽  
Mrna Expression ◽  
Oxidative Dna Damage ◽  
Umbilical Vein ◽  
University Teaching ◽  
Mitochondrial Mass ◽  
Control Subjects ◽  
Mitochondrial Dna Damage ◽  
And Control

Abstract Background Sepsis is a life-threatening condition accompanied by organ dysfunction subsequent to a dysregulated host response to infection. Up to 60% of patients with sepsis develop acute kidney injury (AKI), which is associated with a poor clinical outcome. The pathophysiology of sepsis-associated AKI (sepsis-AKI) remains incompletely understood, but mitochondria have emerged as key players in the pathogenesis. Therefore, our aim was to identify mitochondrial damage in patients with sepsis-AKI. Methods We conducted a clinical laboratory study using “warm” postmortem biopsies from sepsis-associated AKI patients from a university teaching hospital. Biopsies were taken from adult patients (n = 14) who died of sepsis with AKI at the intensive care unit (ICU) and control patients (n = 12) undergoing tumor nephrectomy. To define the mechanisms of the mitochondrial contribution to the pathogenesis of sepsis-AKI, we explored mRNA and DNA expression of mitochondrial quality mechanism pathways, DNA oxidation and mitochondrial DNA (mtDNA) integrity in renal biopsies from sepsis-AKI patients and control subjects. Next, we induced human umbilical vein endothelial cells (HUVECs) with lipopolysaccharide (LPS) for 48 h to mimic sepsis and validate our results in vitro. Results Compared to control subjects, sepsis-AKI patients had upregulated mRNA expression of oxidative damage markers, excess mitochondrial DNA damage and lower mitochondrial mass. Sepsis-AKI patients had lower mRNA expression of mitochondrial quality markers TFAM, PINK1 and PARKIN, but not of MFN2 and DRP1. Oxidative DNA damage was present in the cytosol of tubular epithelial cells in the kidney of sepsis-AKI patients, whereas it was almost absent in biopsies from control subjects. Oxidative DNA damage co-localized with both the nuclei and mitochondria. Accordingly, HUVECs induced with LPS for 48 h showed an increased mnSOD expression, a decreased TFAM expression and higher mtDNA damage levels. Conclusion Sepsis-AKI induces mitochondrial DNA damage in the human kidney, without upregulation of mitochondrial quality control mechanisms, which likely resulted in a reduction in mitochondrial mass.

scholarly journals Asbestos induces mitochondrial DNA damage and dysfunction linked to the development of apoptosis

2003 ◽  
Vol 285 (5) ◽  
pp. L1018-L1025 ◽  
Author(s):  
Arti Shukla ◽  
Michael Jung ◽  
Maria Stern ◽  
Naomi K. Fukagawa ◽  
Douglas J. Taatjes ◽  
...  
Keyword(s):  
Dna Damage ◽  
Mitochondrial Dna ◽  
Cell Injury ◽  
Nuclear Dna ◽  
Oxidant Stress ◽  
Mesothelial Cells ◽  
Confocal Scanning Laser Microscopy ◽  
Mrna Levels ◽  
Mitochondrial Dna Damage ◽  
Induced Apoptosis

To test the hypothesis that asbestos-mediated cell injury is mediated through an oxidant-dependent mitochondrial pathway, isolated mesothelial cells were examined for mitochondrial DNA damage as determined by quantitative PCR. Mitochondrial DNA damage occurred at fourfold lower concentrations of crocidolite asbestos compared with concentrations required for nuclear DNA damage. DNA damage by asbestos was preceded by oxidant stress as shown by confocal scanning laser microscopy using MitoTracker Green FM and the oxidant probe Redox Sensor Red CC-1. These events were associated with dose-related decreases in steady-state mRNA levels of cytochrome c oxidase, subunit 3 (COIII), and NADH dehydrogenase 5. Subsequently, dose-dependent decreases in formazan production, an indication of mitochondrial dysfunction, increased mRNA expression of pro- and antiapoptotic genes, and increased numbers of apoptotic cells were observed in asbestos-exposed mesothelial cells. The possible contribution of mitochondrial-derived pathways to asbestos-induced apoptosis was confirmed by its significant reduction after pretreatment of cells with a caspase-9 inhibitor. Apoptosis was decreased in the presence of catalase. Last, use of HeLa cells transfected with a mitochondrial transport sequence targeting the human DNA repair enzyme 8-oxoguanine DNA glycosylase to mitochondria demonstrated that asbestos-induced apoptosis was ameliorated with increased cell survival. Studies collectively indicate that mitochondria are initial targets of asbestos-induced DNA damage and apoptosis via an oxidant-related mechanism.

scholarly journals p53 is required for nuclear but not mitochondrial DNA damage-induced degeneration

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Matthew J. Geden ◽  
Selena E. Romero ◽  
Mohanish Deshmukh
Keyword(s):  
Dna Damage ◽  
Mitochondrial Dna ◽  
Nuclear Dna ◽  
Neuronal Cell ◽  
Axon Degeneration ◽  
Whole Cell ◽  
Axon Pruning ◽  
Mtdna Damage ◽  
Mitochondrial Dna Damage ◽  
Neuronal Cell Bodies

AbstractWhile the consequences of nuclear DNA damage have been well studied, the exact consequences of acute and selective mitochondrial DNA (mtDNA) damage are less understood. DNA damaging chemotherapeutic drugs are known to activate p53-dependent apoptosis in response to sustained nuclear DNA damage. While it is recognized that whole-cell exposure to these drugs also damages mtDNA, the specific contribution of mtDNA damage to cellular degeneration is less clear. To examine this, we induced selective mtDNA damage in neuronal axons using microfluidic chambers that allow for the spatial and fluidic isolation of neuronal cell bodies (containing nucleus and mitochondria) from the axons (containing mitochondria). Exposure of the DNA damaging drug cisplatin selectively to only the axons induced mtDNA damage in axonal mitochondria, without nuclear damage. We found that this resulted in the selective degeneration of only the targeted axons that were exposed to DNA damage, where ROS was induced but mitochondria were not permeabilized. mtDNA damage-induced axon degeneration was not mediated by any of the three known axon degeneration pathways: apoptosis, axon pruning, and Wallerian degeneration, as Bax-deficiency, or Casp3-deficiency, or Sarm1-deficiency failed to protect the degenerating axons. Strikingly, p53, which is essential for degeneration after nuclear DNA damage, was also not required for degeneration induced with mtDNA damage. This was most evident when the p53-deficient neurons were globally exposed to cisplatin. While the cell bodies of p53-deficient neurons were protected from degeneration in this context, the axons farthest from the cell bodies still underwent degeneration. These results highlight how whole cell exposure to DNA damage activates two pathways of degeneration; a faster, p53-dependent apoptotic degeneration that is triggered in the cell bodies with nuclear DNA damage, and a slower, p53-independent degeneration that is induced with mtDNA damage.

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