高浓度花生四烯酸诱导ECV304细胞凋亡

目的 :探讨花生四烯酸是否能诱导人膀胱癌细胞ECV30 4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。结果 :ECV30 4细胞在 80～ 1 6 0 μmol·L-1 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 1 6 0 μmol·L-1 花生四烯酸处理 2 4h后的ECV30 4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 :80～ 1 6 0 μmol·L-1 的高浓度花生四烯酸能诱导ECV30 4细胞凋亡 ,脂质过氧化作用参与了花生四烯酸致ECV30 4细胞损伤及凋亡过程

Higher Concentration of Arachidonic Acid Induces Apoptosis in ECV304 cells

Objective: To observe whether arachidonic acid (AA) could induce apoptosis in human bladder carcinoma cells ECV304 and the potential mechanism involved. Methods: Cell viability was monitored by MTT assay. The release of lactate dehydrogenase (LDH) was determined by colorimetric assay. Lipid peroxidation in ECV304 cells was measured as malondialdehyde (MDA) content. Hoechst 33258 dying was used to observe AA induced morphological changes. Agarose gel electrophoresis was used to detect DNA fragmentation. Results: Treatment of ECV304 cells with AA for 24 h, in the range of 40～160 μmol·L -1 , caused a great decrease in cell viability,an increase in MDA contents and a release of LDH simultaneously(P<0.01). Cells treated by 160 μmol·L -1 AA showed typical morphological changes of apoptosis. A “ladder” pattern representing DNA fragmentation was also observed after 24 h of AA treatment. Conclusion: Higher concentration of arachidonic acid (80～160 μmol·L -1 ),could induce apoptosis in ECV304 cells. Lipid peroxidation might play a role in AA induced apoptosis of ECV304 cells.