Plant-derived neuroprotective peptides and protein hydrolysates have gained increasing attention for their po-tential in alleviating neurocognitive dysfunction.However,limited research has explored the effects of alca...Plant-derived neuroprotective peptides and protein hydrolysates have gained increasing attention for their po-tential in alleviating neurocognitive dysfunction.However,limited research has explored the effects of alcalase-hydrolyzed black soybean protein hydrolysates(Alc-BSPEHs)and their peptides on cognitive impairment.In this study,we investigated the neuroprotective effects and underlying mechanisms of Alc-BSPEHs on scopolamine-induced memory impairment in mice.Additionally,a target peptide(S-8-F)was identified using traditional purification methods(fast protein liquid chromatography,FPLC)combined with computational screening,and its protective effects were evaluated in scopolamine-treated PC12 cells in vitro.Oral administration of Alc-BSPEHs(666 mg/kg)significantly improved behavioral performance and alleviated cholinergic dysfunction,as evidenced by a 53.29%increase in acetylcholine levels and a 36.35%reduction in acetylcholinesterase(AChE)activity.Alc-BSPEHs also mitigated oxidative stress in the brain,increasing SOD and GSH-px activity by 62.92%and 273.16%,respectively,and reducing MDA levels by 85.94%compared to scopolamine-treated controls.Among the FPLC-purified fractions,F6 exhibited the strongest antioxidant(DPPH:52.31%)and AChE inhibitory(32.19%)activities.Furthermore,molecular docking predicted 14 peptides with high Pepti-deRanker scores(>0.5).Among them,S-8-F showed high binding affinities for Keap1 and AChE,with binding energies of-8.9 and-7.9 kcal/mol,respectively.Molecular dynamics(MD)simulations further confirmed the stability of these complexes.Finally,cellular assays demonstrated S-8-F(0.04 mg/mL)exerted neuroprotective effects in scopolamine-induced PC12 cells by reducing intracellular ROS and AChE activity,increasing acetyl-choline levels,and activating the Nrf2/HO-1 signaling pathway.These findings highlight Alc-BSPEHs,particu-larly peptide S-8-F,as a promising strategy for mitigating neurodegenerative disorders.展开更多
基金supported by the National Key Research and Development Plan(2023YFD2100803)the National Natural Science Foundation of China(32372387,32402015)+5 种基金the Heilongjiang Province Outstanding Youth Fund Project(JQ2024C003)the Natural Science Foundation of Heilongjiang Province(PL2024C011)the Double Firstclass Discipline Collaborative Innovation Achievement Project(LJGXCG202080,LJGXCG202083)the Heilongjiang Province Key Research and Development Plan Unveiling Project(2023ZXJ08B03)the Heilongjiang Province Young Science and Technology Talent Lift Project(2023QNTJ001)the Central Leading Local Science and Technology Development Project(ZY2022B-HRB-12).
文摘Plant-derived neuroprotective peptides and protein hydrolysates have gained increasing attention for their po-tential in alleviating neurocognitive dysfunction.However,limited research has explored the effects of alcalase-hydrolyzed black soybean protein hydrolysates(Alc-BSPEHs)and their peptides on cognitive impairment.In this study,we investigated the neuroprotective effects and underlying mechanisms of Alc-BSPEHs on scopolamine-induced memory impairment in mice.Additionally,a target peptide(S-8-F)was identified using traditional purification methods(fast protein liquid chromatography,FPLC)combined with computational screening,and its protective effects were evaluated in scopolamine-treated PC12 cells in vitro.Oral administration of Alc-BSPEHs(666 mg/kg)significantly improved behavioral performance and alleviated cholinergic dysfunction,as evidenced by a 53.29%increase in acetylcholine levels and a 36.35%reduction in acetylcholinesterase(AChE)activity.Alc-BSPEHs also mitigated oxidative stress in the brain,increasing SOD and GSH-px activity by 62.92%and 273.16%,respectively,and reducing MDA levels by 85.94%compared to scopolamine-treated controls.Among the FPLC-purified fractions,F6 exhibited the strongest antioxidant(DPPH:52.31%)and AChE inhibitory(32.19%)activities.Furthermore,molecular docking predicted 14 peptides with high Pepti-deRanker scores(>0.5).Among them,S-8-F showed high binding affinities for Keap1 and AChE,with binding energies of-8.9 and-7.9 kcal/mol,respectively.Molecular dynamics(MD)simulations further confirmed the stability of these complexes.Finally,cellular assays demonstrated S-8-F(0.04 mg/mL)exerted neuroprotective effects in scopolamine-induced PC12 cells by reducing intracellular ROS and AChE activity,increasing acetyl-choline levels,and activating the Nrf2/HO-1 signaling pathway.These findings highlight Alc-BSPEHs,particu-larly peptide S-8-F,as a promising strategy for mitigating neurodegenerative disorders.
