A Small Dose of Hydrogen Peroxide Enhances Tumor Necrosis Factor-Alpha Toxicity in Inducing Human Vascular Endothelial Cell Apoptosis: Reversal with Propofol

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Abstract
Ot treated, treated with 10 μM of hydrogen peroxide (H2O2), treated with TNF-α (40 ng/mL) alone, TNF-α in the presence of 10 μM of H2O2 (H+T), or propofol plus H2O2 for 24 h. Cell viability was measured by lactate dehydrogenate (LDH) assay. Cell apoptosis was assessed by flow cytometry and terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick end-labeling. The antiapoptotic Bcl-2 and pro-apoptotic Bax protein expressions were measured by immunocytochemical analysis. Increases in apoptosis, Bax, lipid peroxidation product malondialdehyde, LDH, and decreases in Bcl-2, superoxide dismutase, and glutathione peroxidase were observed in TNF-α–treated cells. H2O2 10 μM did not cause significant lipid peroxidation (0.75 ± 0.03 nmol/mg of malondialdehyde protein) as compared with control (0.70 ± 0.04 nmol/mg of malondialdehyde protein) (P > 0.05) but further enhanced TNF-α–induced lipid peroxidation, upregulated Bax, and down-regulated Bcl-2 expression and enhanced TNF-α–induced cell apoptosis (P < 0.05). Propofol 50 μM attenuated TNF-α and H2O2-induced cell apoptosis, accompanied by decreases in malondialdehyde and LDH production and restoration of Bcl-2 expression. Propofol exerts protective effects against H2O2-enhanced TNF-α cell toxicity by reducing oxidative injury....

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