Abstract:To investigate the protective effects of kaempferol-3-O-rutinoside (K3R) against H2O2-induced oxidative damage in HepG2 cells, the regulatory mechanism was systematically analyzed by an in vitro model. Following pre-treatment with 10, 20, and 30 μmol·L-1 K3R for 24 h, the cells were exposed to 400 μmol·L-1 H2O2 for 4 h. Cellular viability, reactive oxygen species levels, oxidative damage status, and antioxidant capacity were assessed across experimental groups. Apoptosis was evaluated using Annexin V-FITC/PI staining and TUNEL assays. Western blotting was performed to analyze the expressions of antioxidant proteins (HO-1, NQO1), apoptosis-related proteins (Bax, Caspase3, Bcl-2), and key components of the MAPK/NF-κB signaling pathway. The results demonstrated that cell viability was significantly increased to 99.93% in the K3R-protected groups, accompanied by reduced ROS levels approaching normal ranges, enhanced antioxidant enzyme activity, and effective suppression of apoptosis. The protein expression of HO-1, NQO1, and Bcl-2 was upregulated, whereas Bax and Caspase3 expression was downregulated. Furthermore, phosphorylation of P38, JNK, ERK1/2, and NF-κB p65 in the MAPK/NF-κB pathway was effectively inhibited. In summary, K3R alleviates H2O2-induced oxidative damage in HepG2 cells by suppressing MAPK/NF-κB pathway activation, enhancing the antioxidant capacity and regulating apoptosis-related proteins. It provides a theoretical basis for its application in the development of antioxidant functional foods and drugs.
