高浓度花生四烯酸诱导小鼠成纤维细胞L929凋亡的研究  

Induction of apoptosis in mouse fibroblast cell line L929 by arachidonic acid

作　　者：王韻[1] 汪炳华[1] 代赵明[1] 洪嘉玲[1] 陈丽达[1] 蔡卫斌[1] 

机构地区：[1]武汉大学医学院生物化学与分子生物学系,湖北武汉430071

出　　处：《中国病理生理杂志》2003年第6期825-828,共4页Chinese Journal of Pathophysiology

摘　　要：目的 :探讨花生四烯酸是否能诱导小鼠成纤维细胞L92 9凋亡及其可能机制。方法 :噻唑蓝 (MTT)法检测细胞存活率 ,比色法测定乳酸脱氢酶 (LDH)释放率 ,硫代巴比妥酸法测定细胞脂质过氧化产物丙二醛 (MDA)含量 ,Hoechst 332 5 8染色观察凋亡细胞核形态变化 ,DNA琼脂糖凝胶电泳检测DNA降解。结果 :L92 9细胞在 4 0 - 16 0μmol/L花生四烯酸作用 2 4h后 ,细胞存活率明显下降 ,LDH释放率和细胞MDA含量显著增加 (P <0 0 1)。经 16 0μmol/L花生四烯酸处理后的L92 9细胞呈现典型的凋亡核固缩表现。琼脂糖凝胶电泳显示DNA凋亡梯带。 结论 :高浓度花生四烯酸 (80 - 16 0 μmol/L)能诱导小鼠成纤维细胞L92 9凋亡 ,可能与其促脂质过氧化作用有关。AIM: To observe whether arachidonic acid (AA) could induce apoptosis in mouse fibroblast cell line L929 and the potential mechanism involved. METHODS: The viability and damaged degree of L929 was monitored by MTT and the release of lactate dehydrogenase (LDH). Lipid peroxidation in L929 was measured as malondialdehyde (MDA) content by colorimetric assay. Hoechst 33258 staining was used to observe AA-induced morphological changes. Agarose gel electrophoresis was used to detect DNA fragmentation. RESULTS: Treatment of L929 cell with AA for 24 h, in the range of 40-160 μmol/L, caused a great decrease in cell survival and increased MDA contents and the release of LDH simultaneously( P< 0.01). Cells treated by 160 μmol/L AA showed typical morphological changes of apoptosis. A 'ladder' pattern representing fragmentation of DNA into oligonucleosome length fragments was observed 24 h after AA treatment. CONCLUSION: Higher concentration of arachidonic acid (80-160 μmol/L) induced apoptosis in mouse fibroblast cell line L929. The mechanism of its action might be related to lipid peroxidation.

关 键 词：花生四烯酸类 成纤维细胞 细胞凋亡 脂质过氧化作用 

分 类 号：R363[医药卫生—病理学]