Wang Bin, Zhong Shuping, Zheng Fuchun, Zhang Yanmei, Gao Fenfei, Chen Yicun, Lu Binger, Xu Han, Shi Ganggang
Department of Pharmacology, Shantou University Medical College, Shantou, 515041 Guangdong, China.
Department of Biochemistry and Molecular Biology, University of Southern California, Los Angeles, CA 90033, USA.
Oncotarget. 2015 Sep 22;6(28):24709-21. doi: 10.18632/oncotarget.5077.
N-n-butyl haloperidol iodide (F2), a novel compound derived from haloperidol, protects against the damaging effects of ischemia/reperfusion (I/R) injury in vitro and in vivo. In this study, we hypothesized the myocardial protection of F2 on cardiomyocyte hypoxia/reoxygenation (H/R) injury is mediated by inhibiting autophagy in H9c2 cells. The degree of autophagy by treatment with F2 exposed to H/R in H9c2 cell was characterized by monodansylcadaverine, transmission electron microscopy, and expression of autophagy marker protein LC3. Our results indicated that treatment with F2 inhibited autophagy in H9c2 cells exposed to H/R. 3-methyladenine, an inhibitor of autophagy, suppressed H/R-induced autophagy, and decreased apoptosis, whereas rapamycin, a classical autophagy sensitizer, increased autophagy and apoptosis. Mechanistically, macrophage migration inhibitory factor (MIF) was inhibited by F2 treatment after H/R. Accordingly, small interfering RNA (siRNA)-mediated MIF knockdown decreased H/R-induced autophagy. In summary, F2 protects cardiomyocytes during H/R injury through suppressing autophagy activation. Our results provide a new mechanistic insight into a functional role of F2 against H/R-induced cardiomyocyte injury and death.
碘化N-正丁基氟哌啶醇（F2）是一种从氟哌啶醇衍生而来的新型化合物，在体外和体内均能保护细胞免受缺血/再灌注（I/R）损伤。在本研究中，我们假设F2对心肌细胞缺氧/复氧（H/R）损伤的心肌保护作用是通过抑制H9c2细胞中的自噬来介导的。用单丹磺酰尸胺、透射电子显微镜以及自噬标记蛋白LC3的表达来表征F2处理H9c2细胞后暴露于H/R时的自噬程度。我们的结果表明，F2处理可抑制暴露于H/R的H9c2细胞中的自噬。自噬抑制剂3-甲基腺嘌呤可抑制H/R诱导的自噬，并减少细胞凋亡，而经典的自噬激活剂雷帕霉素则增加自噬和细胞凋亡。从机制上讲，H/R后F2处理可抑制巨噬细胞迁移抑制因子（MIF）。因此，小干扰RNA（siRNA）介导的MIF敲低可减少H/R诱导的自噬。总之，F2通过抑制自噬激活在H/R损伤期间保护心肌细胞。我们的结果为F2抗H/R诱导的心肌细胞损伤和死亡的功能作用提供了新的机制见解。
