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机构地区:[1]湖南医科大学分子药理研究室
出 处:《中国药理学报》1996年第5期467-470,共4页Acta Pharmacologica Sinica
摘 要:氧化型低密度脂蛋白诱导巨噬细胞凋亡牛喜林,张修武,郭兆贵(湖南医科大学分子药理研究室,长沙410078,中国)关键词DNA片断;细胞凋亡;低密度脂蛋白;腹腔巨噬细胞目的:研究氧化型低密度脂蛋白(oxLDL)诱导巨噬细胞凋亡.方法:超速离心法分离人血...IM: To examine whether oxidized low density lipoproteins (ox LDL) might induce apoptosis in mouse peritoneal macrophages (MPM). METHODS: Low density lipoproteins (LDL) were isolated from healthy human plasma by ultracentrifugation and oxidized by CuSO 4 10 μmol·L -1 . MPM were incubated in a medium containing ox LDL, LDL, or phosphate buffer solution (PBS) as control. DNA fragmentation was visualized by agarose gel electrophoresis and determined quantitatively using Hoechst 33258 fluorochrome. RESULTS: Ox LDL, not LDL, elicited typical apoptotic morphological changes (shrinkage of cytoplasm and condensation of chromatin) and DNA fragmentation in a time and dose dependent manner. Incubation for 24 h was necessary for ox LDL 200 mg protein ·L -1 to induce DNA fragmentation, and the maximal effect reached at 72 h. The DNA fragmentation after 24 h incubation with ox LDL at concentrations of 100, 200, and 400 mg protein·L -1 amounted to 6 0 % ( P >0 05), 9 3 % ( P <0 05), and 30 9 % ( P <0 01) , respectively vs PBS control. Dextran sulfate, a scavenger receptor blocker and cycloheximide, a protein synthesis inhibitor, exhibited no effect on DNA fragmentation. However, antioxidant butylated hydroxytoluene (BHT) 20 μmol·L -1 completely inhibited Cu 2+ mediated oxidation of LDL as well as the apoptosis inducing effect of Cu 2+ exposed LDL. Lysophosphatidylcholine (LPC), an active component in ox LDL, at concentration up to 60 μmol·L -1 , did not elicit DNA fragmentation in MPM. CONCLUSION: Ox LDL induces apoptosis in MPM without involving LPC.
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