NOX4/ROS mediate ethanol‑induced apoptosis via MAPK signal pathway in L‑02 cells

Abstract

The aim of the present study was to assess the molecular mechanism of ethanol‑induced oxidative stress‑mediated apoptosis in L‑02 liver cells in order to elucidate novel pathways associated with alcoholic liver disease. L‑02 cells were treated with 400 mM ethanol with or without inhibitors. The cell viability was measured by an MTT assay. Cell apoptosis was assessed by flow cytometry and a single‑stranded DNA (ssDNA) assay. Intracellular reactive oxygen species (ROS) production of L‑02 cells was determined using the 2',7'‑dichlorofluorescein‑diacetate dye. The protein expression of c‑Jun N‑terminal kinase (JNK), phosphorylated (p)‑JNK, P38, p‑P38, NADPH oxidase (NOX)1, NOX4, p22phox, B‑cell lymphoma 2 (Bcl‑2) and Bcl‑2‑associated X protein were measured by western blot analysis. The mRNA expression of NOX1, NOX4 and p22phox was measured by reverse transcription polymerase chain reaction analysis. The results indicated that after treatment with various concentrations of ethanol for the indicated durations, L‑02 cells were displayed a significant decrease in cell viability in a dose‑and time‑dependent manner. Ethanol‑induced apoptosis and cell death of L‑02 cells was accompanied by the generation of ROS, elevated expression of NOX, as well as phosphorylation of JNK and P‑38. In addition, increased expression of Bcl‑2 was induced by 400 mM ethanol. Furthermore, treatment with NOX inhibitor attenuated the ethanol‑induced a decrease in cell viability, and an increase in apoptosis and Bcl‑2 expression. In conclusion, ethanol induced apoptosis in the L‑02 hepatocyte cell line via generation of ROS and elevated expression of NOX4. This indicated that activation of JNK and p38 in the mitogen‑activated protein kinase pathway promotes apoptosis in L‑02 cells.

Dipeptide YA is Responsible for the Positive Effect of Oyster Hydrolysates on Alcohol Metabolism in Single Ethanol Binge Rodent Models

“…Although the method of displaying concentration is different, 100 ~ 200 mM EtOH induces ~30% cell death. In line with our study, cell death is induced by ~50% in human normal liver cells L-02 treated with 400 mM EtOH for 24 h [ 41 ]. The 2% EtOH used in this study is a very high concentration with 50% cytotoxicity.…”

“…In general, 100 mM is used to induce cell injury. However, even in reports, 100 mM EtOH induces cell death approximately 25% of the time [ 40 , 41 ]. These results are similar to our results.…”

miR-378b Regulates Insulin Sensitivity by Targeting Insulin Receptor and p110α in Alcohol-Induced Hepatic Steatosis

“…Numerous studies have shown that alcohol can cause alcoholic liver damage by activating oxidative stress, inhibiting liver fat-acid metabolism, and increasing liver lipid synthesis (Li et al, 2017;Li et al, 2018;Yang C. F. et al, 2018). Studies have also revealed that alcohol reduces hepatic glycogen synthesis and increases gluconeogenesis by regulating the liver insulin signaling pathway (Pang et al, 2009;Magdaleno et al, 2017).…”

NOX4 activation is involved in ROS‐dependent Jurkat T‐cell death induced by Entamoeba histolytica

“…In addition, NOX4 participation in the stress‐induced human islet cell death process has been demonstrated using a NOX4 inhibitor . Also, it has been reported that elevated NOX4 expression by ethanol induced ROS generation, which causes cell death in hepatocytes . It has also been shown that NOX4 also participated in the death of TGF‐β–induced hepatocytes .…”