摘要
目的 研究小剂量(100 nmol/L)H_2O_2对大鼠肝卵圆细胞株WB-F344细胞膜理化特性、脂质过氧化及基因组DNA的影响。方法利用标记膜脂及膜蛋白质的荧光标记物1，6-二苯基-1，3，5-己三烯(DPH)和N-(3芘)马来酰亚胺[N-(3P)M]标记不同剂量H_2O_2作用及经H_2O_2多次作用后发生转化的WB细胞，通过测定其荧光偏振度确定膜理化特性的改变；以硫代巴比妥酸(TBA)法测定其脂质过氧化产物丙二醛（MDA）含量的变化；以及利用溴化乙锭(EB)作荧光探针，通过DNA-EB结合物荧光强度的变化确定其基因组DNA的损伤程度。结果小剂量(100 nmol/L)H2O2的一次作用可使膜蛋白运动度增加，膜脂流动性增大，MDA的含量增高；H_2O_2的多次作用使上述变化更为显著。大剂量(1 mmol/L)、中剂量(100 μmol/L)的H_2O_2作用细胞后可直接损伤基因组DNA，小剂量H2O2的一次作用虽然不能直接损伤基因组DNA，但多次作用后却可使基因组DNA受损。结论H_2O_2可诱使WB细胞膜理化特性发生改变，脂质过氧化产生及造成基因组DNA损伤，此可能与H_2O_2的致、促癌作用有关。
Objective To study the alterations of membrane physicochemical properties, lipid peroxidation and DNA damage in WB-F344 rat liver oval cell induced by hydrogen peroxide(H_2O_2). Methods The physicochemical properties of cell membrane, malondialdehyde(MDA) content and DNA-EB complex were measured by means of detecting fluorescence polarizations of 1,6-dipheny-1,3,5-hexatriene(DPH) and N-(3-pyrene) maleimide [N-(3P)M] labeled in membrane, thiobarbituric acide (TBA) assay, and fluorescence intensity assay respectively. Results The low concentration of H_2O_2 (100 nmol/L) increased the fluidity of lipid domain and the mobility of protein domain in WB cell membrane. When H_2O_2 reacted with whole WB cell, the amount of MDA increased and fluorescence intensity of DNA-EB complex decreased which meant lipid peroxidation and DNA damage of cell respectively. Conclusions The proliferation and transformation of the cell could be related to the changes of physicochemical properties in the membrane lipid and protein domains, lipid peroxidation of the membrane and DNA damage of the genomics.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2001年第3期247-250,共4页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金(39670196)资助&&
