Overexpression of Aldose Reductase Render Mouse Hepatocytes More Sensitive to Acetaminophen Induced Oxidative Stress and Cell Death

Abstract

Acetaminophen (APAP) a commonly used drug for decrease the fever and pain but is capable to induced hepatotoxicity at over dose. This study was carried out to investigate the effect of APAP on the expression of antiapoptotic and antioxidative defense genes, and whether aldose reductase over-expressing plasmid capable to protect against APAP-induced oxidative stress and cell death. APAP treatment induced oxidative stress and hepatotoxicity, and significantly increased aldose reductase mRNA and protein expression in mouse hepatocyte (AML-12).Unexpectedly, AML-12 cells over-expressing aldose reductase augmented APAP-induced reduction in cell viability, reactive oxygen species (ROS) production, glutathione (GSH) depletion and glutathione S-transferase A2 expression. Moreover, over-expression of aldose reductase potentiated APAP induced reduction on proliferating cell nuclear antigen, B cell lymphoma-extra large (bcl-x L ), catalase, glutathione peroxidase-1 (GPx-1) and abolished APAP-induced B-cell lymphoma 2 (bcl-2) inductions. Further, over-expression of aldose reductase significantly abolished AMP activated protein kinase (AMPK) activity in APAP-treated cells and induced p53 expression. This results demonstrate that APAP induced toxicity in AML-12, increased aldose reductase expression, and over-expression of aldose reductase render this cell more susceptible to APAP induced oxidative stress and cell death, this probably due to inhibition AMPK or bcl-2 activity, or may due to competition between aldose reductase and glutathione reductase for NADPH.

The role of apoptosis in acetaminophen hepatotoxicity

“…More recently, it became popular to measure Bcl-2 mRNA and protein levels during APAP hepatotoxicity (Ahmed et al, 2016; Cao et al, 2018; Dong et al, 2014; Hu et al, 2017; Hong et al, 2012; Li et al, 2013; Sharma et al, 2011; Song et al, 2014; Wang et al, 2017, Wang et al, 2018; Zhang et al, 2017; Zhao et al, 2012). Interestingly, most of these reports show high Bcl-2 expression in controls and declining levels during APAP toxicity (Ahmed et al, 2016; Cao et al, 2018; Dong et al, 2014; Hu et al, 2017; Hong et al, 2012; Li et al, 2013; Sharma et al, 2011; Song et al, 2014; Wang et al, 2017, Wang et al, 2018; Zhang et al, 2017; Zhao et al, 2012). Treatment with natural products appears to reverse this decline (Ahmed et al, 2016; Cao et al, 2018; Dong et al, 2014; Hu et al, 2017; Hong et al, 2012; Li et al, 2013; Sharma et al, 2011; Song et al, 2014; Wang et al, 2017, Wang et al, 2018; Zhang et al, 2017; Zhao et al, 2012).…”

“…Based on the fact that tBid supports the formation of Bax pores, it can be assumed that deficiency of Bid will at best cause a temporary delay in DNA fragmentation and injury similar to Bax deficiency (Bajt et al, 2008). APAP has been shown to reduce Bcl-xL expression and a number of natural products have attenuated this reduction in Bcl-xL expression (Ahmed et al, 2016; Dong et al, 2014; Ray et al, 1999, 2006; Ray and Jena, 2000). However, these results are merely correlative and do not support any causative role of Bcl-xL in APAP-induced cell death.…”

“…However, during the last few years a rapidly increasing number of studies, especially in the natural product literature, are being published that again claim that apoptosis is an important part of the pathophysiology (e.g, Ahmed et al, 2016; Cao et al, 2018; Dong et al, 2014; Hu et al, 2017; Hong et al, 2012; Li et al, 2013; Sharma et al, 2011; Song et al, 2014; Wang et al, 2010; Wang et al, 2017, Wang et al, 2018; Zhang et al, 2017; Zhao et al, 2012). Although most of these studies do not appear to directly challenge previous reports regarding the lack of apoptosis, they mainly ignore the pertinent literature and simply conclude based on the measurement of a few, assumed apoptosis parameters that the natural product that is being tested is protecting due to its anti-apoptotic effect (e.g.…”

“…Although most of these studies do not appear to directly challenge previous reports regarding the lack of apoptosis, they mainly ignore the pertinent literature and simply conclude based on the measurement of a few, assumed apoptosis parameters that the natural product that is being tested is protecting due to its anti-apoptotic effect (e.g. Ahmed et al, 2016; Cao et al, 2018; Dong et al, 2014; Hu et al, 2017; Hong et al, 2012; Li et al, 2013; Sharma et al, 2011; Song et al, 2014; Wang et al, 2010; Wang et al, 2017, Wang et al, 2018; Zhang et al, 2017; Zhao et al, 2012). Unfortunately, this is perpetuating a fundamentally wrong conclusion, which not only questions the proposed mechanism of protection of a particular natural product but runs the risk that many other studies follow their lead.…”

Prediction and mechanistic analysis of Drug-Induced Liver Injury (DILI) based on chemical structure

“…Given that the focus is set on proteins with bioactivity related to DILI risk, we only further examined those that were significantly enriched in DILI-related compounds as determined by a Wilcoxon rank test. This included aldose reductase AKR1B1 which has been linked to APAP-induced oxidative stress and hepatotoxicity (29), the CYP enzymes CYP1A2 and CYP2C9 which are involved in xenobiotic metabolism in the liver (30), and the p38 kinase MAPK11 which is known to mediate stress-related signals in hepatotoxicity (31). Moreover, aldo-keto reductase family 1 member C (AKR1C3) is essential for Phase II drug metabolism pathways and Hypoxia-inducible factor prolyl 4-hydroxylase (P4HTM)…”