Cytoprotection of rat hepatocytes by desipramine in a model of simulated ischemia/reperfusion

• Published in: Biochemistry and biophysics reports 2021-09, Vol.27, p.101075-101075, Article 101075
• Main Authors: Shin, Jun-Kyu, Kim, Jae-Sung
• Format: Article
• Language: English
• Subjects: Autophagy; Desipramine; Ischemia and reperfusion injury; Liver
• ISSN: 2405-5808; 2405-5808
• DOI: 10.1016/j.bbrep.2021.101075

We investigated the cytoprotective effect of desipramine (DMI) during in vitro simulated ischemia/reperfusion (I/R) of rat hepatocytes. Primary hepatocytes isolated from male Sprague-Dawley rats were subjected to 4 h of anoxia at pH 6.2 followed by normoxia at pH 7.4 for 2 h to simulate ischemia and reperfusion, respectively. During simulated reperfusion, some hepatocytes were reoxygenated using media containing 5 μM DMI. Necrotic cell death and the onset of mitochondrial permeability transition (MPT) were assessed using fluorometry and confocal microscopy. Changes in autophagic flux and autophagy-related proteins (ATGs) were analyzed by immunoblotting. DMI was shown to substantially delay MPT onset and suppress I/R related cell damage. Mechanistically, DMI treatment during reperfusion increased the expression level of the microtubule-associated protein 1A/1B-light chain 3 (LC3) processing enzymes, ATG4B and ATG7. Genetic knockdown of ATG4B abolished the cytoprotective effect of DMI. Together, these results indicate that DMI is a unique agent which enhances LC3 processing in an ATG4B-dependent way. •DMI suppresses I/R injury by blocking the onset of mitochondrial permeability transition.•DMI Increases autophagy and mitophagy.•Cytoprotection by DMI is ATG4B-dependent.