Institute of Developmental Biology, School of Life Science, Shandong University, Jinan 250100, China.
Chem Res Toxicol. 2009 Mar 16;22(3):471-5. doi: 10.1021/tx8002824.
We previously found a butyrolactone derivative, 3-benzyl-5-((2-nitrophenoxy) methyl)-dihydrofuran-2(3H)-one (3BDO), could inhibit vascular endothelial cell (VEC) apoptosis and senescence induced by a deprivation of serum and FGF-2. In this study, we aimed to investigate its actions in VEC autophagy induced by chloroquine (CQ). The measurement on the volume of acidic compartments (VAC) and autophagy analysis by acridine orange (AO) staining and microtubule-associated protein 1 light chain 3 (MAP1LC3) process revealed that 3BDO was an effective inhibitor of autophagic vesicle accumulation (vacuolation) induced by CQ in VECs. 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) was used for mitochondrial membrane potential (MMP) measurement. The results showed that CQ elevated MMP significantly and that 3BDO could significantly inhibit CQ-induced MMP increase. Na+,K+-ATPase activity assay showed that CQ inhibited this enzyme activity significantly and that 3BDO attenuated the alteration of Na+,K+-ATPase activity caused by CQ. We concluded that 3BDO was a promising inhibitor of CQ-induced accumulation of autophagic vesicles in VECs and could weaken the alterations of MMP and Na+,K+-ATPase activity induced by CQ. The data indicate that 3BDO will be a potential tool for investigating the mechanism of autophagy.
我们之前发现一种丁内酯衍生物，3-苄基-5-((2-硝基苯氧基)甲基)-二氢呋喃-2(3H)-酮（3BDO），可抑制血清和 FGF-2 剥夺诱导的血管内皮细胞（VEC）凋亡和衰老。在这项研究中，我们旨在研究其在氯喹（CQ）诱导的 VEC 自噬中的作用。通过吖啶橙（AO）染色和微管相关蛋白 1 轻链 3（MAP1LC3）过程测量酸性区室（VAC）的体积和自噬分析表明，3BDO 是 VEC 中 CQ 诱导的自噬囊泡积累（空泡化）的有效抑制剂。5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑基羰花青碘化物（JC-1）用于测量线粒体膜电位（MMP）。结果表明，CQ 显著升高 MMP，3BDO 可显著抑制 CQ 诱导的 MMP 升高。Na+，K+-ATPase 活性测定表明，CQ 显著抑制该酶活性，3BDO 减轻 CQ 引起的 Na+，K+-ATPase 活性改变。我们得出结论，3BDO 是一种有前途的 CQ 诱导 VEC 自噬囊泡积累抑制剂，可减弱 CQ 诱导的 MMP 和 Na+，K+-ATPase 活性改变。数据表明，3BDO 将成为研究自噬机制的潜在工具。
