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Cell Stress and Chaperones

, Volume 14, Issue 2, pp 207–217 | Cite as

Heat-shock response protects peripheral blood mononuclear cells (PBMCs) from hydrogen peroxide-induced mitochondrial disturbance

  • Han-Yao Chiu
  • Lon-Yen Tsao
  • Rei-Cheng Yang
Original Paper

Abstract

The present study was designed to investigate ex vivo the protective mechanisms of heat-shock response against H2O2-induced oxidative stress in peripheral blood mononuclear cells (PBMCs) of rats. Twenty-four hours later, heat-shock treatment was executed in vivo; rat PBMCs were collected and treated with H2O2. The accumulation of reactive oxygen species and the mitochondrial membrane potential were evaluated by intracellular fluorescent dHE and JC-1 dye staining, respectively, and expression of HSP72 and cytochrome c was detected by Western blot analysis. Cellular apoptosis was assayed by TUNEL staining and double staining of Annexin V and PI. The results showed that H2O2-induced oxidative stress leads to intracellular superoxide accumulation and collapse of the mitochondrial membrane potential in rat PBMCs. Moreover, cellular apoptosis was detected after H2O2 treatment, and the release of mitochondrial cytochrome c from mitochondria to cytosol was significantly enhanced. Heat-shock pretreatment decreases the accumulation of intracellular superoxide in PBMCs during H2O2-induced oxidative stress. Moreover, heat-shock treatment prevents the collapse of the mitochondrial membrane potential and cytochrome c release from mitochondria during H2O2-induced oxidative stress. In conclusion, mitochondria are critical organelles of the protective effects of heat-shock treatment. Cellular apoptosis during H2O2-induced oxidative stress is decreased by heat-shock treatment through a decrease in superoxide induction and preservation of the mitochondrial membrane potential.

Keywords

Heat-shock response Rat Superoxide Oxidative stress Mitochondrion 

Abbreviation

DAB

diaminobenzidine

dHE

dihydroethidium

HSP

heat-shock protein

Δψ

mitochondrial membrane potential

PBMC

peripheral blood mononuclear cell

ROS

Reactive oxygen species

PI

propidium iodium

TUNEL

TdT-mediated dUTP nick end-labeling assay

TdT

terminal deoxynucleotidyl transferase

JC-1

5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazol-carbocyanine iodide

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Copyright information

© Cell Stress Society International 2008

Authors and Affiliations

  1. 1.Department of Respiratory CareChang Jung Christian UniversityTainanTaiwan
  2. 2.Department of PediatricsChanghua Christian HospitalChanghuaTaiwan
  3. 3.Department of Pediatrics, College of MedicineKaohsiung Medical UniversityKaohsiung CityTaiwan

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