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Results of cardiopulmonary resuscitation (CPR) for asphyxial cardiac arrest (aCA). The CPR time, epinephrine used during CPR, survival rate and neurological deficit (ND) scores in rats, were compared in saline and histidine-tryptophan-ketoglutarate solution (HTK) groups. (A) CPR time (p < 0.05 for the 6'30@ groups (endotracheal tube clamped and asphyxia for 6 min 30 s) compared to the 4'30@ groups (endotracheal tube clamped and asphyxia for 4 min 30s)). (B) The epinephrine dose used for CPR. (C) Survival rate on day 3. ( � p < 0.05, HTK vs. saline in the 6 0 30@ group). (D) Total neurological deficit (ND) scores are shown from day 0 to day 3. Numbers listed beside each icon on the line chart are the survived and observed rats at that time point. Treatment with HTK improved ND scores significantly at day 1 for the 4 0 30@ group and from day 1 to day 3 for the 6 0 30@ group. ( � p < 0.05 compared with the saline-treated controls with the same duration time of aCA). https://doi.org/10.1371/journal.pone.0221039.g005
Source publication
Ischemic neuron loss contributes to brain dysfunction in patients with cardiac arrest (CA). Histidine-tryptophan-ketoglutarate (HTK) solution is a preservative used during organ transplantation. We tested the potential of HTK to protect neurons from severe hypoxia (SH) following CA. We isolated rat primary cortical neurons and induced SH with or wi...
Contexts in source publication
Context 1
... was used because it could increase arterial pressure and coronary perfusion pressure, and provide good successful resuscitation rate while low dosage was used [24,25]. The 4 0 30@ groups (asphyxia for 4 min and 30 s) had shorter durations of resuscitation, and the 6 0 30@ groups (asphyxia for 6 min and 30 s) were resuscitated significantly longer (p < 0.05 as compared to the 4 0 30@ groups with the same treatment), but the difference in resuscitation duration between the saline and HTK groups was not significant at the same asphyxia time (Fig 5A). For the epinephrine dosage, the 6 0 30@ groups used nearly twice the dosage as the 4 0 30@ groups. ...
Context 2
... the epinephrine dosage, the 6 0 30@ groups used nearly twice the dosage as the 4 0 30@ groups. No significant differences were observed between the HTK group and the saline group with the same asphyxia time (Fig 5B). ...
Context 3
... significance (p = 0.6547), and significantly from 26.7% to 44.4% for the 6 0 30@ group at day 3 (p < 0.05, Fig 5C). The infarction size of the 4 0 30@ groups was lower than that of the 6 0 30@ groups, and was lower in the groups treated with HTK than that of saline-treated groups; however, these were not statistically significant. ...
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Numerous studies have demonstrated that metformin can reduce the incidence of myocardial infarction and improve the prognosis of patients. However, its specific mechanism has not been determined. Using a rat model of myocardial ischemia‑reperfusion injury (MIRI), it was observed that metformin significantly reduced infarct size, and decreased the l...
Citations
... Seven different isoforms of the NOX family have been identified, NOX1 to NOX5, as well as dual oxidase 1 (DUOX1) and DUOX2, each with similar but distinct structural and functional characteristics [1]. NOX4 is considered to be expressed in most cell types and regulated by altered gene expression, hypoxia, and the polymerase (DNA-directed) deltainteracting protein 2 [2][3][4][5]. At physiological conditions, NOX4, by producing hydrogen peroxide and superoxide, affects cell signaling events such as those promoted by receptor tyrosine kinases [6]. ...
Previous studies have reported beneficial effects of NADPH oxidase 4 (NOX4) inhibition on beta-cell survival in vitro and in vivo. The mechanisms by which NOX4 inhibition protects insulin producing cells are, however, not known. The aim of the present study was to investigate the effects of a pharmacological NOX4 inhibitor (GLX7013114) on human islet and EndoC-βH1 cell mitochondrial function, and to correlate such effects with survival in islets of different size, activity, and glucose-stimulated insulin release responsiveness. We found that maximal oxygen consumption rates, but not the rates of acidification and proton leak, were increased in islets after acute NOX4 inhibition. In EndoC-βH1 cells, NOX4 inhibition increased the mitochondrial membrane potential, as estimated by JC-1 fluorescence; mitochondrial reactive oxygen species (ROS) production, as estimated by MitoSOX fluorescence; and the ATP/ADP ratio, as assessed by a bioluminescent assay. Moreover, the insulin release from EndoC-βH1 cells at a high glucose concentration increased with NOX4 inhibition. These findings were paralleled by NOX4 inhibition-induced protection against human islet cell death when challenged with high glucose and sodium palmitate. The NOX4 inhibitor protected equally well islets of different size, activity, and glucose responsiveness. We conclude that pharmacological alleviation of NOX4-induced inhibition of beta-cell mitochondria leads to increased, and not decreased, mitochondrial ROS, and this was associated with protection against cell death occurring in different types of heterogeneous islets. Thus, NOX4 inhibition or modulation may be a therapeutic strategy in type 2 diabetes that targets all types of islets.
... A series of culture medium samples (200 μL) were collected after treatment. The culture medium was immediately kept in the dark and on ice until chemiluminescence (CL) measurement, usually finished within 2 h, as described previously [54,56]. Before CL measurement, 0.1 mL of phosphate-buffered saline (pH 7.4) was added to 0.1 mL of culture medium. ...
... CL was monitored continuously for an additional 300 s. We previously showed that the CL recordings of lucigenin and luminol are derived from superoxide and H2O2, respectively [54,56]. For superoxide, the total amount of CL was calculated by integrating the area under the curve and subtracting it from the background level, expressed as CL counts per second. ...
... RNA was extracted using a commercial kit (RareRNA, Bio-East Technology, Taipei, Taiwan), as previously described [56], and cDNA was synthesized at 42 °C for 45 min (reaction mixture: 2 μg of RNA, 5 μg of poly(dT)15 oligonucleotide primer (Life Technologies, Waltham, MA, USA)) and exposed to 200 units of reverse transcriptase (Moloney murine leukemia virus; Promega, Madison, WI, USA). Quantitative RT-PCR was performed in an ABI StepOne Plus system (Applied Biosystems, Foster City, CA, USA). ...
Inflammation worsens oxalate nephropathy by exacerbating tubular damage. The transient receptor potential vanilloid 1 (TRPV1) channel is present in kidney and has a polymodal sensing ability. Here, we tested whether TRPV1 plays a role in hyperoxaluria-induced renal inflammation. In TRPV1-expressed proximal tubular cells LLC-PK1, oxalate could induce cell damage in a time- and dose-dependent manner; this was associated with increased arachidonate 12-lipoxygenase (ALOX12) expression and synthesis of endovanilloid 12(S)-hydroxyeicosatetraenoic acid for TRPV1 activation. Inhibition of ALOX12 or TRPV1 attenuated oxalate-mediated cell damage. We further showed that increases in intracellular Ca2+ and protein kinase C α activation are downstream of TRPV1 for NADPH oxidase 4 upregulation and reactive oxygen species formation. These trigger tubular cell inflammation via increased NLR family pyrin domain-containing 3 expression, caspase-1 activation, and interleukin (IL)-1β release, and were alleviated by TRPV1 inhibition. Male hyperoxaluric rats demonstrated urinary supersaturation, tubular damage, and oxidative stress in a time-dependent manner. Chronic TRPV1 inhibition did not affect hyperoxaluria and urinary supersaturation, but markedly reduced tubular damage and calcium oxalate crystal deposition by lowering oxidative stress and inflammatory signaling. Taking all these results together, we conclude that TRPV1 hyperfunction contributes to oxalate-induced renal inflammation. Blunting TRPV1 function attenuates hyperoxaluric nephropathy.
... Stroke was one of the leading causes of disability and the second highest cause of death globally, with certain obviously increasing prevalence in some areas [1], while ischemic stroke is accounted for approximately 72.5% of strokes overall [2]. Of note, stroke was the main cause of neurological function loss and neuronal death during the process of cerebral ischemic injury [3]. Brain reperfusion injury following ischemic stroke was defined as damage of brain tissue suffered from insufficient blood supply which dramatically reduced the benefits of new establishing blood flow for acute ischemic stroke [4]. ...
Introduction:
The advances in cerebral ischemia treatment have resulted in a larger proportion of patients get the benefits of rebuilding blood flow to the brain. Then, ischemia-reperfusion injury has emerged as a new essential problem. Dectin-1 plays an important role in cerebral ischemia-reperfusion injury by regulating the function of immune cells.
Methods:
C57BL/6 was blindly divided into four groups including the sham-operated group and the three different kinds of middle cerebral artery occlusion (MCAO) groups (after 6 hours, 12 hours, and 24 hours after plug removal). The protein expression levels of dectin-1, proapoptosis molecule, and antiapoptosis molecule were measured by using western blot analysis. The brain tissue was analyzed by flow cytometry to detect the M1 macrophage levels.
Results:
The correlation analysis of dectin-1 and infarct areas showed that there was an obviously positive correlation in between them (R = 0.9603). Dectin-1, cleaved caspase-3, and Bax increased, while antiapoptosis molecule, Bcl-2, decreased at three appropriate time points (after 6 hours, 12 hours, and 24 hours). The level of M1 macrophages in the experimental group increased after ischemia-reperfusion injury compared to the control group.
Conclusions:
The high expression level of dectin-1 may affect M1 macrophage polarization and brain cell apoptosis in cerebral ischemia-reperfusion injury.
... The absorbance of the mixture solution was measured at 492 nm. The concentration of LDH was quantitated against a standard protein obtained from Sigma-Aldrich, previously described [54][55][56]. The cells' viability was determined using a 3-(4,5)-2,5-diphenyltetrazolium bromide (MTT) method. ...
... The optical density (O.D.) was measured at 570 nm to determine cell viability. The cell viability was calculated as viable cells (%) = (O.D. of treated sample/O.D. of control sample) × 100, as previously described [54][55][56]. ...
... Cellular RNA was isolated using a commercial kit (RareRNA, Bio-East Technology, Taipei, Taiwan), as previously described [56]. The DNase I amplification kit (Invitrogen, Carlsbad, CA, USA) was added to eliminate genomic DNA in RNA solution. ...
Indoxyl sulfate (IS) is accumulated during severe renal insufficiency and known for its nephrotoxic properties. Transient receptor potential vanilloid 1 (TRPV1) is present in the kidney and acts as a renal sensor. However, the mechanism underlying IS-mediated renal tubular damage in view of TRPV1 is lacking. Here, we demonstrated that TRPV1 was expressed in tubular cells of Lilly Laboratories cell-porcine kidney 1 (LLC-PK1) and Madin-Darby canine kidney cells (MDCK). IS treatment in both cells exhibited tubular damage with increased LDH release and reduced cell viability in dose- and time-dependent manners. MDCK, however, was more vulnerable to IS. We, therefore, investigated MDCK cells to explore a more detailed mechanism. Interestingly, IS-induced tubular damage was markedly attenuated in the presence of selective TRPV1 blockers. IS showed no effect on TRPV1 expression but significantly increased arachidonate 12-lipoxygenase (ALOX12) protein, mRNA expression, and 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE) amounts in a dose-dependent manner, indicating that the ALOX12/12(S)-HETE pathway induced TRPV1 hyperfunction in IS-mediated tubulotoxicity. Blockade of ALOX12 by cinnamyl-3,4-dihydroxy-α-cyanocinnamate or baicalein attenuated the effects of IS. Since aryl hydrocarbon receptor (AhR) activation after IS binding is crucial in mediating cell death, here, we found that the AhR blockade not only ameliorated tubular damage but also attenuated ALOX12 expression and 12(S)-HETE production caused by IS. The uremic toxic adsorbent AST-120, however, showed little effect on ALOX12 and 12(S)-HETE, as well as IS-induced cell damage. These results clearly indicated that IS activated AhR and then upregulated ALOX12, and this induced endovanilloid 12(S)-HETE synthesis and contributed to TRPV1 hyperfunction in IS-treated tubular cells. Further study on TRPV1 may attenuate kidney susceptibility to the functional loss of end-stage kidney disease via IS.
Subarachnoid hemorrhage (SAH), a type of hemorrhagic stroke, is a neurological emergency associated with a high morbidity and mortality rate. After SAH, early brain injury (EBI) is the leading cause of poor prognosis in SAH patients. Peroxiredoxins (PRDXs) are a family of sulphhydryl-dependent peroxidases. Peroxiredoxin-3 (PRDX3) is mainly located in the mitochondria of neurons, which can remove hydrogen peroxide (H2O2); however, the effect of PRDX3 on EBI after SAH remains unclear. In this study, an endovascular perforation model was used to mimic SAH in Sprague Dawley rats in vivo. The results revealed that after SAH, PRDX3 levels decreased in the neurons. PRDX3 overexpression by neuron-specific adeno-associated viruses upregulated PRDX3 levels. Furthermore, PRDX3 overexpression improved long- and short-term behavioral outcomes and alleviated neuronal impairment in rats. Nissl staining revealed that the upregulation of PRDX3 promoted cortical neuron survival. PRDX3 overexpression decreased the H2O2 content and downregulated caspase-9 expression. In conclusion, PRDX3 participates in neuronal protection by inhibiting the neuronal mitochondria-mediated death pathway; PRDX3 may be an important target for EBI intervention after SAH.
Hydrogen peroxide (H2O2), as a common disinfectant, has been extensively used in aquaculture. The toxicity of high ambient H2O2 for gills and liver of fish has received attention from many researchers. However, whether H2O2 exposure induced brain injury and neurotoxicity has not been reported in fish. Therefore, this study aimed to explore the potential mechanism of H2O2 toxicity in brain of common carp via transcriptome analysis and biochemical parameter detection. The fish were exposed to 0 (control) and 1 mM of H2O2 for 1 h per day lasting 14 days. The results showed that H2O2 exposure caused oxidative damage in brain evidenced by decreased glutathione (GSH), total antioxidant capacity (T-AOC) and nicotinamide adenine dinucleotide (NAD⁺) levels, and increased formation of malondialdehyde (MDA) and 8-hydroxy-2′-deoxyguanosine (8-OHdG). Meanwhile, H2O2 exposure reduced 5-hydroxytryptamine (5-HT) level, and down-regulated tryptophan hydroxylase 1 (tph1a), tph2, 5-hydroxytryptamine receptor 1A-beta (htr1ab) and htr2b expression in brain. Transcriptome analysis showed that H2O2 exposure up-regulated 604 genes and down-regulated 1209 genes in brain. Go enrichment displayed that the differently expressed genes (DEGs) were enriched mainly in cellular process, single-organism process, metabolic process, and biological regulation in the biological process category. Further, KEGG enrichment indicated that H2O2 exposure led to dysregulation of neurotransmitter signals including depression of glutamatergic synapse, GABAergic synapse and endocannabinoid signaling. Also, we found the alteration of three key pathways including calcium, cAMP and HIF-1 in brain after H2O2 exposure. In conclusion, our data indicated that H2O2 exposure induced oxidative damage and neurotoxicity, possibly related to dysregulation of neurotransmitters and calcium, cAMP and HIF-1 signaling pathways, which may adversely affect learning, memory and social responses of common carp. This study provided novel insight into biological effects and underlying mechanism of H2O2 toxicity in aquatic animal, and contributed to proper application of H2O2 in aquaculture.
Glutamate N-methyl-D-aspartate receptor (NMDAR) hyperfunction is known to contribute to acute renal failure due to ischemia/reperfusion and endotoxemia. D-Serine is a co-agonist for NMDAR activation, but whether NMDARs play a role in D-serine-mediated nephrotoxicity remains unclear. Here, we demonstrate that NMDAR blockade ameliorates D-serine-induced renal injury. In NMDAR-expressing LLC-PK 1 cells, which were used as a proximal tubule model, D-serine but not L-serine induced cytotoxicity in a dose-dependent manner, which was abrogated by selective NMDAR blockers MK-801 and AP-5. Time-dependent oxidative stress, evidenced by gradually increased superoxide and H 2 O 2 production, was associated with D-serine-mediated cytotoxicity; these reactive oxygen species could be alleviated not only after NMDAR inhibition but also by NADPH oxidase (NOX) inhibition. Activation of protein kinase C (PKC)d and PKCz is a downstream signal for NMDAR-mediated NOX activation because PKC inhibition diminishes the NOX activity that is induced by D-serine. Renal injury was further confirmed in male Wistar rats that intraperitoneally received D-serine but not L-serine. Peak changes in glucosuria, proteinuria, and urinary excretion of lactate dehydrogenase and malondialdehyde were found after 24 h of treatment. Persistent tubular damage was observed after 7 days of treatment. Co-treatment of the NMDAR blocker MK-801 for 24 h abolished D-serine-induced functional insufficiency and tubular damage. MK-801 attenuated renal superoxide formation by lowering NOX activity and protein upregulation of NOX4 but not NOX2. These results reveal that NMDAR hyperfunction underlies D-serine-induced renal injury via the effects of NOX4 on triggering oxidative stress.
