Yadav Umesh C S, Ramana K V, Srivastava Satish K
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555, USA.
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555, USA.
Free Radic Biol Med. 2013 Dec;65:15-25. doi: 10.1016/j.freeradbiomed.2013.06.008. Epub 2013 Jun 12.
Aldose reductase (AR), a glucose-metabolizing enzyme, reduces lipid aldehydes and their glutathione conjugates with more than 1000-fold efficiency (Km aldehydes 5-30 µM) relative to glucose. Acrolein, a major endogenous lipid peroxidation product as well as a component of environmental pollutants and cigarette smoke, is known to be involved in various pathologies including atherosclerosis, airway inflammation, COPD, and age-related disorders, but the mechanism of acrolein-induced cytotoxicity is not clearly understood. We have investigated the role of AR in acrolein-induced cytotoxicity in primary human small airway epithelial cells (SAECs). Exposure of SAECs to varying concentrations of acrolein caused cell death in a concentration- and time-dependent manner. AR inhibition by fidarestat prevented the low-dose (5-10 µM) but not the high-dose (>10 µM) acrolein-induced SAEC death. AR inhibition protected SAECs from low-dose (5 µM) acrolein-induced cellular reactive oxygen species (ROS). Inhibition of acrolein-induced apoptosis by fidarestat was confirmed by decreased condensation of nuclear chromatin, DNA fragmentation, comet tail moment, and annexin V fluorescence. Further, fidarestat inhibited acrolein-induced translocation of the proapoptotic proteins Bax and Bad from the cytosol to the mitochondria and that of Bcl2 and BclXL from the mitochondria to the cytosol. Acrolein-induced cytochrome c release from mitochondria was also prevented by AR inhibition. The mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinases 1 and 2, stress-activated protein kinase/c-Jun NH2-terminal kinase, and p38MAPK, and c-Jun were transiently activated in airway epithelial cells by acrolein in a concentration- and time-dependent fashion, which was significantly prevented by AR inhibition. These results suggest that AR inhibitors could prevent acrolein-induced cytotoxicity in the lung epithelial cells.
醛糖还原酶(AR)是一种葡萄糖代谢酶,相对于葡萄糖,它能以超过1000倍的效率(醛的Km值为5 - 30 μM)还原脂质醛及其谷胱甘肽共轭物。丙烯醛是一种主要的内源性脂质过氧化产物,也是环境污染物和香烟烟雾的成分之一,已知其与多种病理状况有关,包括动脉粥样硬化、气道炎症、慢性阻塞性肺疾病(COPD)和与年龄相关的疾病,但丙烯醛诱导细胞毒性的机制尚不清楚。我们研究了AR在人原代小气道上皮细胞(SAECs)中丙烯醛诱导的细胞毒性中的作用。将SAECs暴露于不同浓度的丙烯醛会导致细胞死亡,且呈浓度和时间依赖性。非达司他抑制AR可预防低剂量(5 - 10 μM)但不能预防高剂量(>10 μM)丙烯醛诱导的SAEC死亡。AR抑制可保护SAECs免受低剂量(5 μM)丙烯醛诱导的细胞活性氧(ROS)产生。非达司他抑制丙烯醛诱导的凋亡可通过核染色质凝聚减少、DNA片段化、彗星尾矩和膜联蛋白V荧光的降低得到证实。此外,非达司他抑制了丙烯醛诱导的促凋亡蛋白Bax和Bad从细胞质向线粒体的转位以及Bcl2和BclXL从线粒体向细胞质的转位。AR抑制还可预防丙烯醛诱导的细胞色素c从线粒体释放。丝裂原活化蛋白激酶(MAPK),如细胞外信号调节激酶1和2、应激激活蛋白激酶/c-Jun氨基末端激酶以及p38MAPK,和c-Jun在气道上皮细胞中被丙烯醛以浓度和时间依赖性方式短暂激活,而AR抑制可显著预防这种激活。这些结果表明,AR抑制剂可预防丙烯醛诱导的肺上皮细胞毒性。