Endoplasmic reticulum (ER) stress and unfolded protein response are closely associated with liver injury and liver disease progression, but molecular mechanisms are not fully understood. We have discovered several novel molecular targets for ER stress-induced hepatocyte injury. The expression of pleckstrin homology-like domain, family A, member-3 (PHLDA3) was induced by ER stress in hepatocytes, which was confirmed in patients and animal models with liver diseases. The increase in PHLDA3 by ER stress resulted from IRE1-dependent Xbp1s activation, which leads to Akt inhibition responsible for liver injury. In addition, the beneficial effect of farnesoid X receptor (FXR) ER stress-mediated NLRP3 inflammasome activation was revealed. FXR activation inhibited PERK-CHOP signaling pathway through the regulation of miR-186. Moreover, dual-specificity phosphatase 5 (DUSP5) was discovered as a novel target. ER stress increased DUSP5 expression in hepatocytes through CHOP activation, contributing to ERK inhibition and hepatocyte injury. Overall, these findings may provide the potential targets for ER stress-associated hepatocyte dysfunction in various pathological conditions.