Introduction:Human chemerin is an adipokine that regulates chemotaxis,inflammation,and glucose metabolism.In addition,accumulating evidence suggests that chemerin promotes apoptosis,autophagy,and pyroptosis.However,th...Introduction:Human chemerin is an adipokine that regulates chemotaxis,inflammation,and glucose metabolism.In addition,accumulating evidence suggests that chemerin promotes apoptosis,autophagy,and pyroptosis.However,there are no data on its impact on eryptosis.The current study aimed to analyze the effects of human active Glu^(21)-Ser^(157) chemerin on eryptosis in vitro.Materials and Methods:Human chemerin 0-2-10-50μg/mL was incubated for 24 h with human erythrocytes(hematocrit 0.4%)obtained from eight healthy individuals.Flow cytometry-based determination of phospholipid scrambling,reactive oxygen species(ROS)production,and intracellular Ca^(2+)levels was performed.To supplement data on ROS and Ca^(2+)signaling in chemerin-mediated eryptosis,incubation in the presence or absence of antioxidants vitamin C and N-acetylcysteine and Ca^(2+)-binding agent EGTA was carried out,respectively.Confocal microscopy-based techniques were used to detect reactive nitrogen species(RNS)generation,involvement of caspase-3 and caspase-8,as well as the state of lipid order in cell membranes of erythrocytes exposed to human Glu^(21)-Ser^(157) chemerin.Results:Our observations suggest that human Glu^(21)-Ser^(157) chemerin had no impact on eryptosis parameters at 2μg/mL.However,chemerin stimulated phosphatidylserine externalization,ROS production,and Ca^(2+)accumulation at higher concentrations suggesting activation of eryptosis.Ca^(2+)uptake turned out to be at least partly required for chemerin-mediated eryptosis.Chemerin-mediated erythrotoxicity was additionally mediated by RNS,caspase-3,and caspase-8.Moreover,Glu^(21)-Ser^(157) chemerin promoted reduction in the liquid-ordered phase of cell membranes in erythrocytes.Conclusions:The present study first discloses that human chemerin can induce eryptosis via Ca^(2+)-dependent mechanisms at concentrations noticeably exceeding circulating levels.Thus,chemerin-induced eryptosis can hardly contribute to eryptosis-mediated anemia in diseases associated with enhanced levels of chemerin in blood.展开更多
This paper presents original research studying the effects of biocompatible nanoparticles standardized(ICNB)on the permeability of erythrocytes and eryptosis.The presented evidence demonstrates that changing orientati...This paper presents original research studying the effects of biocompatible nanoparticles standardized(ICNB)on the permeability of erythrocytes and eryptosis.The presented evidence demonstrates that changing orientation and mobility protons of the hydrogen atoms in the pericellular fluid significantly modifies the permeability and physiological activity of erythrocytes.The leading role of the state of cell membrane and transport activity enzymes(ATPase)in ensuring its permeability and functional activity is exaggerated.Magnetite nanoparticles(ICNB)act on the fluid of pericellular structure by changing the orientation and mobility of hydrogen protons that ultimately determine the permeability,and physiological activity of cells.These studies support the G.Ling's theory of an"association-induction"and"multi-layered organization polarized water".展开更多
Objectives:Targeting epigenetic modifications in anticancer therapy is a promising approach to overcoming cancer cell chemoresistance.The histone deacetylase/DNA methyltransferase inhibitor,parthenolide(PTL),has antit...Objectives:Targeting epigenetic modifications in anticancer therapy is a promising approach to overcoming cancer cell chemoresistance.The histone deacetylase/DNA methyltransferase inhibitor,parthenolide(PTL),has antitumor activity,but contrasting findings exist on its effect in normal cells.This study aims to examine the nongenomic toxic mechanisms of PTL in human erythrocytes.Methods:Cell death as stimulated by 20–200μMof PTL for 24 h at 37℃ was assessed using fluorescence-assorted cell sorting and spectrophotometric assays.Canonical markers of cell death,including membrane scrambling,oxidative stress,and Ca^(2+)mobilization,were captured by annexin V-fluorescein isothiocyanate,2′,7′-dichlorodihydrofluorescein diacetate,and Fluo4/AM labeling,respectively.Rescue experiments usingawide arrayof inhibitorswere also conducted.Results:PTL stimulated significantmembrane scrambling and blebbing,and showed potent hemolytic activity coupled with significant elevations in dichlorofluorescein and Fluo4 fluorescence.While hemolysis was ameliorated by glutathione,caffeine,acetylsalicylic acid,staurosporin,necrosulfonamide,guanosine,and Ca^(2+)deprivation,it was rather exacerbated by necrostatin-2,tumor necrosis factor α(TNFα)or Fas ligand(FasL)neutralization,and concurrent Ca^(2+)deprivation and membrane depolarization.In contrast,eryptosis was attenuated by N-acetyl-cysteine,TNFα,or FasL blockade,and simultaneous Ca^(2+)elimination and KCl enrichment;and augmented by necrostatin-2,necrosulfonamide,and adenosine triphosphate.Interestingly,loss of volume was only prevented by melatonin,acetylsalicylic acid,and N(gamma)-nitro-L-arginine methyl ester.Conclusion:PTL stimulates oxidative hemolysis and eryptosis through Ca^(2+)mobilization and death ligand signaling involving the cyclooxygenase/protein kinase C/mixed lineage kinase domain-like pseudokinase axis.This research highlights the non-genomic toxic mechanisms of PTL and presents potential pharmacological targets for mitigating its adverse off-target effects.展开更多
文摘Introduction:Human chemerin is an adipokine that regulates chemotaxis,inflammation,and glucose metabolism.In addition,accumulating evidence suggests that chemerin promotes apoptosis,autophagy,and pyroptosis.However,there are no data on its impact on eryptosis.The current study aimed to analyze the effects of human active Glu^(21)-Ser^(157) chemerin on eryptosis in vitro.Materials and Methods:Human chemerin 0-2-10-50μg/mL was incubated for 24 h with human erythrocytes(hematocrit 0.4%)obtained from eight healthy individuals.Flow cytometry-based determination of phospholipid scrambling,reactive oxygen species(ROS)production,and intracellular Ca^(2+)levels was performed.To supplement data on ROS and Ca^(2+)signaling in chemerin-mediated eryptosis,incubation in the presence or absence of antioxidants vitamin C and N-acetylcysteine and Ca^(2+)-binding agent EGTA was carried out,respectively.Confocal microscopy-based techniques were used to detect reactive nitrogen species(RNS)generation,involvement of caspase-3 and caspase-8,as well as the state of lipid order in cell membranes of erythrocytes exposed to human Glu^(21)-Ser^(157) chemerin.Results:Our observations suggest that human Glu^(21)-Ser^(157) chemerin had no impact on eryptosis parameters at 2μg/mL.However,chemerin stimulated phosphatidylserine externalization,ROS production,and Ca^(2+)accumulation at higher concentrations suggesting activation of eryptosis.Ca^(2+)uptake turned out to be at least partly required for chemerin-mediated eryptosis.Chemerin-mediated erythrotoxicity was additionally mediated by RNS,caspase-3,and caspase-8.Moreover,Glu^(21)-Ser^(157) chemerin promoted reduction in the liquid-ordered phase of cell membranes in erythrocytes.Conclusions:The present study first discloses that human chemerin can induce eryptosis via Ca^(2+)-dependent mechanisms at concentrations noticeably exceeding circulating levels.Thus,chemerin-induced eryptosis can hardly contribute to eryptosis-mediated anemia in diseases associated with enhanced levels of chemerin in blood.
文摘This paper presents original research studying the effects of biocompatible nanoparticles standardized(ICNB)on the permeability of erythrocytes and eryptosis.The presented evidence demonstrates that changing orientation and mobility protons of the hydrogen atoms in the pericellular fluid significantly modifies the permeability and physiological activity of erythrocytes.The leading role of the state of cell membrane and transport activity enzymes(ATPase)in ensuring its permeability and functional activity is exaggerated.Magnetite nanoparticles(ICNB)act on the fluid of pericellular structure by changing the orientation and mobility of hydrogen protons that ultimately determine the permeability,and physiological activity of cells.These studies support the G.Ling's theory of an"association-induction"and"multi-layered organization polarized water".
基金funded by the Ongoing Research Funding Program at King Saud University,Riyadh,Saudi Arabia through grant number ORF-2025-554.
文摘Objectives:Targeting epigenetic modifications in anticancer therapy is a promising approach to overcoming cancer cell chemoresistance.The histone deacetylase/DNA methyltransferase inhibitor,parthenolide(PTL),has antitumor activity,but contrasting findings exist on its effect in normal cells.This study aims to examine the nongenomic toxic mechanisms of PTL in human erythrocytes.Methods:Cell death as stimulated by 20–200μMof PTL for 24 h at 37℃ was assessed using fluorescence-assorted cell sorting and spectrophotometric assays.Canonical markers of cell death,including membrane scrambling,oxidative stress,and Ca^(2+)mobilization,were captured by annexin V-fluorescein isothiocyanate,2′,7′-dichlorodihydrofluorescein diacetate,and Fluo4/AM labeling,respectively.Rescue experiments usingawide arrayof inhibitorswere also conducted.Results:PTL stimulated significantmembrane scrambling and blebbing,and showed potent hemolytic activity coupled with significant elevations in dichlorofluorescein and Fluo4 fluorescence.While hemolysis was ameliorated by glutathione,caffeine,acetylsalicylic acid,staurosporin,necrosulfonamide,guanosine,and Ca^(2+)deprivation,it was rather exacerbated by necrostatin-2,tumor necrosis factor α(TNFα)or Fas ligand(FasL)neutralization,and concurrent Ca^(2+)deprivation and membrane depolarization.In contrast,eryptosis was attenuated by N-acetyl-cysteine,TNFα,or FasL blockade,and simultaneous Ca^(2+)elimination and KCl enrichment;and augmented by necrostatin-2,necrosulfonamide,and adenosine triphosphate.Interestingly,loss of volume was only prevented by melatonin,acetylsalicylic acid,and N(gamma)-nitro-L-arginine methyl ester.Conclusion:PTL stimulates oxidative hemolysis and eryptosis through Ca^(2+)mobilization and death ligand signaling involving the cyclooxygenase/protein kinase C/mixed lineage kinase domain-like pseudokinase axis.This research highlights the non-genomic toxic mechanisms of PTL and presents potential pharmacological targets for mitigating its adverse off-target effects.
