scholarly journals Noninvasive monitoring of hepatic glutathione depletion through fluorescence imaging and blood testing

2021 ◽  
Vol 7 (8) ◽  
pp. eabd9847
Author(s):  
Xingya Jiang ◽  
Qinhan Zhou ◽  
Bujie Du ◽  
Siqing Li ◽  
Yingyu Huang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Fluorescence Imaging ◽  
High Specificity ◽  
Glutathione Depletion ◽  
Entire Body ◽  
Preclinical Research ◽  
Blood Testing ◽  
Surgical Biopsy ◽  
Hepatic Glutathione

Hepatic glutathione plays a key role in regulating redox potential of the entire body, and its depletion is known to increase susceptibility to oxidative stress involved in many diseases. However, this crucial pathophysiological event can only be detected noninvasively with high-end instrumentation or invasively with surgical biopsy, limiting both preclinical research and clinical prevention of oxidative stress–related diseases. Here, we report that both in vivo fluorescence imaging and blood testing (the first-line detection in the clinics) can be used for noninvasive and consecutive monitoring of hepatic glutathione depletion at high specificity and accuracy with assistance of a body-clearable nanoprobe, of which emission and surface chemistries are selectively activated and transformed by hepatic glutathione in the liver sinusoids. These findings open a new avenue to designing exogenous blood markers that can carry information of local disease through specific nanobiochemical interactions back to the bloodstream for facile and rapid disease detection.

scholarly journals Ochratoxin A-Induced Hepatotoxicity through Phase I and Phase II Reactions Regulated by AhR in Liver Cells

Toxins ◽  
2019 ◽  
Vol 11 (7) ◽  
pp. 377 ◽  
Author(s):  
Hye Soo Shin ◽  
Hyun Jung Lee ◽  
Min Cheol Pyo ◽  
Dojin Ryu ◽  
Kwang-Won Lee
Keyword(s):  
Oxidative Stress ◽  
Phase I ◽  
Ochratoxin A ◽  
Phase Ii ◽  
Glutathione Depletion ◽  
Related Factor ◽  
Aryl Hydrocarbon ◽  
Similar Mode ◽  
Phase I And Ii

Ochratoxin A (OTA) is a widespread mycotoxin produced by several species of the genera Aspergillus and Penicillium. OTA exists in a variety of foods, including rice, oats, and coffee and is hepatotoxic, with a similar mode of action as aflatoxin B1. The precise mechanism of cytotoxicity is not yet known, but oxidative damage is suspected to contribute to its cytotoxic effects. In this study, human hepatocyte HepG2 cells were treated with various concentrations of OTA (5–500 nM) for 48 h. OTA triggered oxidative stress as demonstrated by glutathione depletion and increased reactive oxygen species, malondialdehyde level, and nitric oxide production. Apoptosis was observed with 500 nM OTA treatment. OTA increased both the mRNA and protein expression of phase I and II enzymes. The same results were observed in an in vivo study using ICR mice. Furthermore, the relationship between phase I and II enzymes was demonstrated by the knockdown of the aryl hydrocarbon receptor (AhR) and NF-E2-related factor 2 (Nrf2) with siRNA. Taken together, our results show that OTA induces oxidative stress through the phase I reaction regulated by AhR and induces apoptosis, and that the phase II reaction is activated by Nrf2 in the presence of oxidative stress.

scholarly journals PML is a ROS sensor activating p53 upon oxidative stress

2017 ◽  
Vol 214 (11) ◽  
pp. 3197-3206 ◽  
Author(s):  
Michiko Niwa-Kawakita ◽  
Omar Ferhi ◽  
Hassane Soilihi ◽  
Morgane Le Bras ◽  
Valérie Lallemand-Breitenbach ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Biology ◽  
Antioxidant Properties ◽  
Promyelocytic Leukemia ◽  
Nuclear Bodies ◽  
Glutathione Depletion ◽  
Pml Nuclear Bodies ◽  
P53 Activation ◽  
Induced Changes

Promyelocytic leukemia (PML) nuclear bodies (NBs) recruit partner proteins, including p53 and its regulators, thereby controlling their abundance or function. Investigating arsenic sensitivity of acute promyelocytic leukemia, we proposed that PML oxidation promotes NB biogenesis. However, physiological links between PML and oxidative stress response in vivo remain unexplored. Here, we identify PML as a reactive oxygen species (ROS) sensor. Pml−/− cells accumulate ROS, whereas PML expression decreases ROS levels. Unexpectedly, Pml−/− embryos survive acute glutathione depletion. Moreover, Pml−/− animals are resistant to acetaminophen hepatotoxicity or fasting-induced steatosis. Molecularly, Pml−/− animals fail to properly activate oxidative stress–responsive p53 targets, whereas the NRF2 response is amplified and accelerated. Finally, in an oxidative stress–prone background, Pml−/− animals display a longevity phenotype, likely reflecting decreased basal p53 activation. Thus, similar to p53, PML exerts basal antioxidant properties but also drives oxidative stress–induced changes in cell survival/proliferation or metabolism in vivo. Through NB biogenesis, PML therefore couples ROS sensing to p53 responses, shedding a new light on the role of PML in senescence or stem cell biology.

scholarly journals ATF4 is an oxidative stress–inducible, prodeath transcription factor in neurons in vitro and in vivo

2008 ◽  
Vol 205 (5) ◽  
pp. 1227-1242 ◽  
Author(s):  
Philipp S. Lange ◽  
Juan C. Chavez ◽  
John T. Pinto ◽  
Giovanni Coppola ◽  
Chiao-Wang Sun ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Transcription Factor ◽  
Neurological Diseases ◽  
Glutathione Depletion ◽  
Bzip Transcription Factor ◽  
Behavioral Recovery ◽  
Activating Transcription Factor 4 ◽  
Death And Loss

Oxidative stress is pathogenic in neurological diseases, including stroke. The identity of oxidative stress–inducible transcription factors and their role in propagating the death cascade are not well known. In an in vitro model of oxidative stress, the expression of the bZip transcription factor activating transcription factor 4 (ATF4) was induced by glutathione depletion and localized to the promoter of a putative death gene in neurons. Germline deletion of ATF4 resulted in a profound reduction in oxidative stress–induced gene expression and resistance to oxidative death. In neurons, ATF4 modulates an early, upstream event in the death pathway, as resistance to oxidative death by ATF4 deletion was associated with decreased consumption of the antioxidant glutathione. Forced expression of ATF4 was sufficient to promote cell death and loss of glutathione. In ATF4−/− neurons, restoration of ATF4 protein expression reinstated sensitivity to oxidative death. In addition, ATF4−/− mice experienced significantly smaller infarcts and improved behavioral recovery as compared with wild-type mice subjected to the same reductions in blood flow in a rodent model of ischemic stroke. Collectively, these findings establish ATF4 as a redox-regulated, prodeath transcriptional activator in the nervous system that propagates death responses to oxidative stress in vitro and to stroke in vivo.

scholarly journals Highly efficient neuronal gene knockout in vivo by CRISPR-Cas9 via neonatal intracerebroventricular injection of AAV in mice

Gene Therapy ◽  
2021 ◽  
Author(s):  
Sam Hana ◽  
Michael Peterson ◽  
Helen McLaughlin ◽  
Eric Marshall ◽  
Attila J. Fabian ◽  
...  
Keyword(s):  
Gene Knockout ◽  
Low Cost ◽  
High Specificity ◽  
Specific Gene ◽  
Preclinical Research ◽  
Guide Rnas ◽  
Neuronal Gene ◽  
Sgrna Design

AbstractCRISPR-Cas systems have emerged as a powerful tool to generate genetic models for studying normal and diseased central nervous system (CNS). Targeted gene disruption at specific loci has been demonstrated successfully in non-dividing neurons. Despite its simplicity, high specificity and low cost, the efficiency of CRISPR-mediated knockout in vivo can be substantially impacted by many parameters. Here, we used CRISPR-Cas9 to disrupt the neuronal-specific gene, NeuN, and optimized key parameters to achieve effective gene knockout broadly in the CNS in postnatal mice. Three cell lines and two primary neuron cultures were used to validate the disruption of NeuN by single-guide RNAs (sgRNA) harboring distinct spacers and scaffold sequences. This triage identified an optimal sgRNA design with the highest NeuN disruption in in vitro and in vivo systems. To enhance CRISPR efficiency, AAV-PHP.B, a vector with superior neuronal transduction, was used to deliver this sgRNA in Cas9 mice via neonatal intracerebroventricular (ICV) injection. This approach resulted in 99.4% biallelic indels rate in the transduced cells, leading to greater than 70% reduction of total NeuN proteins in the cortex, hippocampus and spinal cord. This work contributes to the optimization of CRISPR-mediated knockout and will be beneficial for fundamental and preclinical research.

The role of oxidative stress in the toxicity of pyridoxal isonicotinoyl hydrazone (PIH) analogues

2002 ◽  
Vol 30 (4) ◽  
pp. 755-758 ◽  
Author(s):  
J. L. Buss ◽  
J. Neuzil ◽  
P. Ponka
Keyword(s):  
Oxidative Stress ◽  
K562 Cells ◽  
Redox Status ◽  
Jurkat Cells ◽  
Glutathione Depletion ◽  
Pyridoxal Isonicotinoyl Hydrazone ◽  
Fe Complexes

Pyridoxal isonicotinoyl hydrazone (PIH) analogues are effective iron chelators in vivo and in vitro, and may be of value for the treatment of secondary iron overload. The sensitivity of Jurkat cells to Fe-chelator complexes was enhanced several-fold by the depletion of the antioxidant glutathione, indicating the role of oxidative stress in their toxicity. K562 cells loaded with eicosapentaenoic acid, a fatty acid particularly susceptible to oxidation, were also more sensitive to the toxic effects of the Fe complexes, and toxicity was proportional to lipid peroxidation. Thus Fechelator complexes cause oxidative stress, which may be a major component of their toxicity. As was the case for their Fe complexes, the toxicity of PIH analogues was enhanced by glutathione depletion of Jurkat cells and eicosapentaenoic acidloading of K562 cells. Thus the toxicity of the chelators themselves is also enhanced by compromised cellular redox status. In addition, the toxicity of the chelators was diminished by culturing Jurkat cells under hypoxic conditions, which may limit the production of the reactive oxygen species that initiate oxidative stress. A significant part of the toxicity of the chelators may be due to intracellular formation of Fe-chelator complexes, which oxidatively destroy the cell.

Recombinant human erythropoietin prevents etoposide- and methotrexate-induced toxicity in kidney and liver tissues via the regulation of oxidative damage and genotoxicity in Wistar rats

2017 ◽  
Vol 37 (8) ◽  
pp. 848-858 ◽  
Author(s):  
K Rjiba-Touati ◽  
I Amara ◽  
M Bousabbeh ◽  
I Ben Salem ◽  
A Azzebi ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Recombinant Human Erythropoietin ◽  
Wistar Rats ◽  
Glutathione Depletion ◽  
Control Group ◽  
Protective Effects ◽  
Drug Induced ◽  
Human Erythropoietin ◽  
Liver Tissues

Etoposide (ETO) and methotrexate (MTX) are two effective chemotherapeutic drugs. However, the clinical use of these drugs is limited by its toxicity in normal tissues, especially in kidney and in liver tissues. Recombinant human erythropoietin (rhEPO), erythropoietin hormone, has also been shown to exert tissue protective effects. The purpose of this study was to explore the protective effect of rhEPO against oxidative stress and genotoxicity induced by ETO and MTX in vivo. Adult male Wistar rats were divided into 10 groups (6 animals each): control group, rhEPO alone group, ETO alone group, MTX alone group and rhEPO + ETO/MTX groups. In rhEPO + ETO/MTX groups, three doses of pretreatment with rhEPO were performed: 1000, 3000 and 6000 IU/kg. Our results showed that rhEPO pretreatment protects liver and kidney tissues against oxidative stress induced by the anticancer drugs. The glycoprotein decreased malondialdehyde (MDA) levels, reduced catalase activity and ameliorated glutathione depletion. Furthermore, we showed that rhEPO administration prevented drug-induced DNA damage accessed by comet test. Altogether, our results suggested a protective role of rhEPO, especially at 3000 IU/kg, against ETO- and MTX-induced oxidative stress and genotoxicity in vivo.

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