Purification and Identification of Peptides from Hydrilla verticillata (Linn. f.) Royle with Cytoprotective
and Antioxidative Effect against H2O2‑Treated
HepG2 Cells
posted on 2024-02-15, 18:05authored byYufang Tang, Fan Liang, Yue Yan, Yanlin Zeng, Yuqin Li, Rong Zhou
Antioxidant
peptides were purified from Hydrilla
verticillata (Linn. f.) Royle (HVR) protein hydrolysate by ultrafiltration, gel filtration chromatography,
and semipreparative reversed-phase HPLC and identified by UPLC-ESI-MS/MS.
Therein, TCLGPK and TCLGER were selected to be synthesized, and they
displayed desirable radical-scavenging activity to ABTS (99.20 ±
0.56–99.20 ± 0.43%), DPPH (97.32 ± 0.59–97.56
± 0.97%), hydroxyl radical (54.32 ± 1.27–70.42 ±
2.01%), and superoxide anion (42.93 ± 1.46–52.62 ±
1.11%) at a concentration of 0.96 μmol/mL. They possessed a
cytoprotective effect against H2O2-induced oxidative
stress in HepG2 cells in a dose-dependent manner. 1.6 μmol/mL
of the two peptides could perfectly protect HepG2 cells from H2O2-induced injury. The TCLGPK exhibited higher
antioxidant activity and cytoprotective effect than TCLGER. Western
blot and molecular docking results indicated that the two peptides
achieved antioxidant ability and cytoprotective effect by combining
with Kelch-like ECH-associated protein 1 (Keap1) to activate the Keap1–nuclear
factor erythroid 2-related factor 2 (Nrf2)–antioxidant response
elements signaling pathway, leading to the activity and expression
of the related antioxidases in the pathway significantly up-regulating
and the intracellular reactive oxygen species level, lipid peroxidation,
and cell apoptosis rate significantly down-regulating.