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ANESTHETIC PHARMACOLOGY

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

From the Department of Anesthesiology, Anesthesiology Research Laboratories, Renmin Hospital of Wuhan University, People's Republic of China.

Address correspondence and reprint requests to Zhengyuan Xia, MD, PhD, Anesthesiology Research Laboratory, Department of Anesthesiology, Renmin Hospital of Wuhan University, Wuhan, 430060 People's Republic of China. Address e-mail to zhengyuan_xia{at}yahoo.com.

We designed the present study to test the hypothesis that oxygen free radicals can enhance tumor necrosis factor (TNF)- cellular toxicity, which might be reversed by propofol, an anesthetic with antioxidant properties, in human vascular endothelial cell line ECV304. Cultured ECV304 were either not treated, treated with 10 µM of hydrogen peroxide (H₂O₂), treated with TNF- (40 ng/mL) alone, TNF- 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-–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-–induced lipid peroxidation, upregulated Bax, and down-regulated Bcl-2 expression and enhanced TNF-–induced cell apoptosis (P < 0.05). Propofol 50 µM attenuated TNF- 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- cell toxicity by reducing oxidative injury.

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Anesthesia & Analgesia® is published for the International Anesthesia Research Society® by Lippincott Williams & Wilkins with the assistance of Stanford University Libraries' HighWire Press®. Copyright 2006 by the International Anesthesia Research Society. Online ISSN: 1526-7598   Print ISSN: 0003-2999