Sertraline induces endoplasmic reticulum stress in hepatic cells

Abstract

Sertraline is used for the treatment of depression, and is also used for the treatment of panic, obsessive-compulsive, and post-traumatic stress disorders. Previously, we have demonstrated that sertraline caused hepatic cytotoxicity, with mitochondrial dysfunction and apoptosis being underlying mechanisms. In this study, we used microarray and other biochemical and molecular analyses to identify endoplasmic reticulum (ER) stress as a novel molecular mechanism. HepG2 cells were exposed to sertraline and subjected to whole genome gene expression microarray analysis. Pathway analysis revealed that ER stress is among the significantly affected biological changes. We confirmed the increased expression of ER stress makers by real-time PCR and Western blots. The expression of typical ER stress markers such as PERK, IRE1α, and CHOP was significantly increased. To study better ER stress-mediated drug-induced liver toxicity; we established in vitro systems for monitoring ER stress quantitatively and efficiently, using Gaussia luciferase (Gluc) and secreted alkaline phosphatase (SEAP) as ER stress reporters. These in vitro systems were validated using well-known ER stress inducers. In these two reporter assays, sertraline inhibited the secretion of Gluc and SEAP. Moreover, we demonstrated that sertraline-induced apoptosis was coupled to ER stress and that the apoptotic effect was attenuated by 4-phenylbutyrate, a potent ER stress inhibitor. In addition, we showed that the MAP4K4-JNK signaling pathway contributed to the process of sertraline-induced ER stress. In summary, we demonstrated that ER stress is a mechanism of sertraline-induced liver toxicity.

Endoplasmic Reticulum Stress Induction and ERK1/2 Activation Contribute to Nefazodone-Induced Toxicity in Hepatic Cells

“…We have previously reported the construction of a Gaussia luciferase reporter assay to monitor ER stress (Chen et al ., 2014b,c). In brief, we established a stable cell line (HepG2-Fluc-Gluc) expressing firefly luciferase (Fluc) and secreted Gaussia luciferase (Gluc).…”

“…The primer information and amplification conditions are detailed in our previous publication (Chen et al ., 2014c). The PCR products, including the XBP1 unspliced (289 bp), spliced (263 bp), and the internal control GAPDH, were analyzed by electrophoresis on 4% (w/v) agarose gels, stained with 0.5 µg/ml ethidium bromide solution, and visualized under UV light.…”

“…The role of ER stress in drug-induced liver toxicity has been under-investigated but has recently received increased attention (Chen et al ., 2014b; Dara et al ., 2011). A number of in vivo and in vitro studies, including ours, has demonstrated that ER stress plays an important role in liver toxicity caused by drugs, such as the analgesic acetaminophen, the antidepressant sertraline, and the anti-HIV1 agent efavirenz (Apostolova et al ., 2013; Chen et al ., 2014c; Fredriksson et al ., 2014; Uzi et al ., 2013). …”

Activation of the Nrf2 signaling pathway in usnic acid-induced toxicity in HepG2 cells

“…cDNA was synthesized from 2 μg of total RNA using a high-capacity cDNA reverse transcription kit (Applied Biosystems, Foster City, CA) according to the manufacturer’s protocol. Real-time PCR was performed as described previously (Chen et al 2014b) to examine the gene expression levels of Nrf2 and GAPDH using the following probes obtained from Applied Biosystems: human Nrf2 (NFE2L2, Hs00975961_g1) and human GAPDH (Hs02758991_g1).…”

Development of HepG2-derived cells expressing cytochrome P450s for assessing metabolism-associated drug-induced liver toxicity

“…The causes of DILI are multifactorial, including toxic effects caused by reactive metabolites, reactive oxygen species, inflammatory reactions, mitochondrial dysfunction, and imbalances between cellular damage and protective responses [1][2][3][4][5]. Metabolism-related toxicity is generally mediated by the generation of reactive metabolites from non-toxic parenteral compounds, particularly via cytochrome P450 (CYP) enzyme pathways [6].…”