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Oligopeptides (Thr–His–Leu–Pro–Lys (THLPK), His–Pro–Leu–Lys (HPLK), Leu–Pro–Lys (LPK), His–Leu–Lys (HLK), and Leu–His–Lys (LHK)) are newly identified from rapeseed napin (Brassica napus) protein-derived hydrolysates with the capability of upregulating glucose transporter-4 (GLUT4) expression and translocation. However, whether each of them enhances GLUT4 expression and translocation and their specific mechanisms remain unclear. Here, we assess the effects of the oligopeptides against insulin resistance (IR) and oxidative stress in hepatocytes and screen out the most antidiabetic one. Specifically, compared with other oligopeptides, LPK not only remarkably elevated glucose consumption to 8.45 mmol/L protein; superoxide dismutase (SOD) activity to 319 U/mg protein; GLUT4 expression and translocation; and phosphorylated level of insulin receptor substrate-1 (IRS-1), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (Akt) (P < 0.05) but also remarkably attenuated the reactive oxygen species (ROS) level to 2255, lactate dehydrogenase (LDH) activity to 20.5 U/mg protein, malondialdehyde (MDA) content to 241 nmol/mg protein, and NO content to 1302 μmol/mL protein (P < 0.05). These findings demonstrated that antidiabetic oligopeptide LPK possessed the most potential to protect HepG2 cells from IR and oxidative stress via activating IRS-1/PI3K/Akt/GLUT4 and regulating common oxidative markers in vitro.