Ginsburg I, Kohen R, Ligumsky M
Department of Oral Biology, Hebrew University-Hadassah School of Dental Medicine, Jerusalem, Israel.
Free Radic Biol Med. 1994 Feb;16(2):263-9. doi: 10.1016/0891-5849(94)90151-1.
Monkey kidney epithelial cells, labeled with chromium and grown in culture, were killed in a synergistic manner when subtoxic amounts of ethanol were combined either with subtoxic amounts of glucose oxidase-generated hydrogen peroxide, or with mixtures of peroxide and with 2,2'-Azo-bis (2-amidinopropane)HCl (AAPH)-generated peroxyl radical. A further enhancement of cytotoxicity occurred when subtoxic amounts of trypsin were added to mixtures of all three agents. While ethanol alone caused shrinkage of the monolayers and cell rounding, no visible cytotoxic changes were observed. Hydrogen peroxide at the concentrations used (about 1 mM), caused only some cell rounding. On the other hand, cells exposed simultaneously to ethanol and to H2O2 developed extensive membrane damage characterized by the formation of large polar blebs, which is compatible with altered membrane permeability. The presence of trypsin markedly enhanced cellular cytotoxicity induced by mixtures of peroxide, peroxyl radical, and ethanol. This could markedly be depressed by catalase and by dimethylthiourea. The tissue culture model described might serve to further investigate the role played by synergy among oxidants and a variety of membrane-damaging agents, and by xenobiotics in tissue damage induced by inflammatory processes.
用铬标记并在培养中生长的猴肾上皮细胞，当亚毒性剂量的乙醇与亚毒性剂量的葡萄糖氧化酶产生的过氧化氢，或与过氧化物和2,2'-偶氮双（2-脒基丙烷）盐酸盐（AAPH）产生的过氧自由基的混合物联合使用时，会以协同方式被杀死。当将亚毒性剂量的胰蛋白酶添加到所有三种试剂的混合物中时，细胞毒性进一步增强。虽然单独的乙醇会导致单层细胞收缩和细胞变圆，但未观察到明显的细胞毒性变化。所用浓度（约1 mM）的过氧化氢仅导致一些细胞变圆。另一方面，同时暴露于乙醇和过氧化氢的细胞出现广泛的膜损伤，其特征是形成大的极性泡，这与膜通透性改变一致。胰蛋白酶的存在显著增强了过氧化物、过氧自由基和乙醇混合物诱导的细胞毒性。这可以被过氧化氢酶和二甲基硫脲显著抑制。所描述的组织培养模型可能有助于进一步研究氧化剂与多种膜损伤剂之间的协同作用以及外源性物质在炎症过程诱导的组织损伤中所起的作用。
