A small dose of hydrogen peroxide enhances tumor necrosis factor-alpha toxicity in inducing human vascular endothelial cell apoptosis: Reversal with propofol
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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DOI：
10.1213/01.ane.0000221183.02244.80
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发表时间：
2006-07-01
影响因子：
5.7
通讯作者：
Xia, Zhengyuan
中科院分区：
文献类型：
--
作者：
Luo, Tao;Xia, Zhengyuan
我们设计了本项研究以验证氧自由基能够增强肿瘤坏死因子（TNF）-α的细胞毒性这一假设，而在人血管内皮细胞系ECV304中，具有抗氧化特性的麻醉剂丙泊酚可能逆转这种毒性。培养的ECV304细胞分为以下几组：未处理组、用10μM过氧化氢（H₂O₂）处理组、单独用TNF -α（40 ng/mL）处理组、在10μM H₂O₂存在的情况下用TNF -α（H + T）处理组，以及丙泊酚加H₂O₂处理24小时组。通过乳酸脱氢酶（LDH）测定法检测细胞活力。通过流式细胞术和末端脱氧核苷酸转移酶（TdT）介导的脱氧尿苷三磷酸（dUTP）缺口末端标记法评估细胞凋亡。通过免疫细胞化学分析检测抗凋亡蛋白Bcl - 2和促凋亡蛋白Bax的表达。在TNF -α处理的细胞中观察到凋亡增加、Bax增加、脂质过氧化产物丙二醛增加、LDH增加，以及Bcl - 2减少、超氧化物歧化酶减少和谷胱甘肽过氧化物酶减少。与对照组（0.70 ± 0.04 nmol/mg丙二醛蛋白）相比，10μM H₂O₂未引起显著的脂质过氧化（0.75 ± 0.03 nmol/mg丙二醛蛋白）（P > 0.05），但进一步增强了TNF -α诱导的脂质过氧化，上调了Bax，下调了Bcl - 2的表达，并增强了TNF -α诱导的细胞凋亡（P < 0.05）。50μM丙泊酚减轻了TNF -α和H₂O₂诱导的细胞凋亡，同时伴有丙二醛和LDH产生减少以及Bcl - 2表达的恢复。丙泊酚通过减轻氧化损伤对H₂O₂增强的TNF -α细胞毒性发挥保护作用。
We designed the present study to test the hypothesis that oxygen free radicals can enhance tumor necrosis factor (TNF)-alpha 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 mu M of hydrogen peroxide (H2O2), treated with TNF-alpha (40 ng/mL) alone, TNF-alpha in the presence of 10 mu M 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. Increase's in apoptosis, Bax, lipid peroxidation product malondialdehyde, LDH, and decreases in Bcl-2, superoxide dismutase, and glutathione peroxidase were observed in TNF-alpha-treated cells. H2O2 10 mu M 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 malondialclehyde protein) (P > 0.05) but further enhanced TNF-alpha-induced lipid peroxidation, upregulated Bax, and down-regulated Bcl-2 expression and enhanced TNF-alpha-induced cell apoptosis (P < 0.05). Propofol 50 mu M attenuated TNF-alpha 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-a cell toxicity by reducing oxidative injury.