目的 研究川芎嗪 (TMP)对花生四烯酸 (AA,16 0μm ol/ L)诱导的血管内皮细胞株 ECV 30 4损伤和凋亡的保护作用及其机制。方法 MTT法检测细胞存活率,比色法测乳酸脱氢酶 (L DH)释放率和细胞丙二醛(MDA)含量,Hoechst 332 5 8荧光染色法观察细胞核形态变化并测定细胞凋亡率,琼脂糖凝胶电泳检测 DNA降解。结果 TMP(0.2 5～1.0 m mol/ L)能浓度依赖性地抑制 AA引起的细胞存活率下降 (P<0.0 5),并降低细胞L DH释放率和 MDA含量 (P<0.0 5)。 AA处理 2 4 h后的细胞呈典型的凋亡核固缩表现,凝胶电泳显示 DNA凋亡梯带,TMP(0.2 5～1.0 m mol/ L)能降低其凋亡率 (P<0.0 5)。结论 TMP对 AA引起的 ECV30 4细胞损伤和凋亡具有保护作用,其机制可能与其抗脂质过氧化作用有关。

Object To study the protective effect of tetramethylpyrazine (TMP) against apoptosis of vascular endothelial cell line ECV304 induced by 160 μmol/L arachidonic acid (AA). Methods The viability and damaged degree of ECV304 cells were monitored by MTT assay. The release of lactate dehydrogenase (LDH) and malondialdehyde (MDA) content in ECV304 cells were measured by colorimetric assay. Hoechst 33258 dyeing was used to observe AA induced morphological changes and count apoptotic ratio. Agarose gel electrophoresis was used to detect DNA fragmentation. Results TMP (0.25-1.0 mmol/L) prevented the inhibitory effect of viability induced by AA (P0.05). TMP (0.25-1.0 mmol/L) partly prevented the increase in the release of LDH and MDA contents induced by AA (P0.05). Cells treated by 160 μmol/L AA showed typical morphological changes of apoptosis. A "DNA ladder" was also observed. TMP (0.25-1.0 mmol/L) partly decreased the ratio of apoptotic cells (P0.05). Conclusion TMP has protective effect on injury and apoptosis of ECV304 cells induced by AA and the mechanism of its action would be related to antilipid peroxidation.

湖北省自然科学基金资助项目(NO₂003ABA188)