Yoon Sun Myung, Lee Ye-Won, Kim Min Ju, Shin Jae-Joon, Bae Gun Won, Park Sunmin
Department of R&D, Unimed Pharmaceuticals Inc., Unimed Bldg., Seoul 05567, Republic of Korea.
Department of Food and Nutrition, Obesity/Diabetes Research Center, Hoseo University, Asan-si 31499, Republic of Korea.
Int J Mol Sci. 2025 Jun 24;26(13):6030. doi: 10.3390/ijms26136030.
This study investigated the potential neuroprotective mechanisms of porcine brain enzyme hydrolysate (PBEH) against Alzheimer's disease pathology using differentiated SH-SY5Y cells. Differentiated neuronal cells were treated with 40 μM amyloid-β(1-42; Aβ) to induce neurotoxicity, followed by PBEH treatment (12.5-400 μg/mL), Com-A (peptide-based neuroprotective supplement; 200 μg/mL) treatment, and Com-B (herbal extract known for improving memory function; 100 μg/mL) treatment. Key assessments included cell viability, Aβ aggregation in adding 10 μM Aβ, amyloidogenic proteins (APP, BACE), synaptic markers (BDNF, ERK), apoptotic markers (BAX/BCL-2, caspase-3), oxidative stress (reactive oxygen species (ROS)), cholinergic function (ChAT, AChE), MAPK signaling (JNK, p38), and neuroinflammation (IL-1β). PBEH contained high concentrations of amino acids, including L-lysine (32.3 mg/g), L-leucine (42.4 mg/g), L-phenylalanine (30.0 mg/g) and the PSIS peptide (86.9 μg/g). Treatment up to 400 μg/mL showed no cytotoxicity and had cognitive protection effects up to 152% under Aβ stress ( < 0.05). PBEH significantly attenuated Aβ aggregation, decreased APP (28%) and BACE (51%) expression, enhanced synaptic function through increased BDNF, and restored ERK phosphorylation ( < 0.05). Anti-apoptotic effects included a 76% reduction in the BAX/BCL-2 ratio, a 47% decrease in caspase-3, and a 56% reduction in ROS levels. Cholinergic function showed restoration via increased ChAT activity ( < 0.01) and decreased AChE activity ( < 0.05). PBEH reduced IL-1β levels by 70% and suppressed JNK/p38 phosphorylation ( < 0.05). While Com-A enhanced BDNF and Com-B showed anti-inflammatory effects, PBEH demonstrated activity across multiple pathway markers. In conclusion, these findings suggest that PBEH may enable neuronal preservation through multi-pathway modulation, establishing foundational evidence for further mechanistic investigation in cognitive enhancement applications.
本研究使用分化的SH-SY5Y细胞，探究了猪脑酶水解物（PBEH）针对阿尔茨海默病病理的潜在神经保护机制。用40μM淀粉样蛋白-β（1-42；Aβ）处理分化的神经元细胞以诱导神经毒性，随后进行PBEH处理（12.5 - 400μg/mL）、Com-A（基于肽的神经保护补充剂；200μg/mL）处理和Com-B（以改善记忆功能而闻名的草药提取物；100μg/mL）处理。关键评估指标包括细胞活力、添加10μM Aβ时的Aβ聚集、淀粉样蛋白生成蛋白（APP、BACE）、突触标记物（BDNF、ERK）、凋亡标记物（BAX/BCL-2、caspase-3）、氧化应激（活性氧（ROS））、胆碱能功能（ChAT、AChE）、MAPK信号传导（JNK、p38）和神经炎症（IL-1β）。PBEH含有高浓度的氨基酸，包括L-赖氨酸（32.3mg/g）、L-亮氨酸（42.4mg/g）、L-苯丙氨酸（30.0mg/g）和PSIS肽（86.9μg/g）。高达400μg/mL的处理未显示细胞毒性，并且在Aβ应激下具有高达152%的认知保护作用（<0.05）。PBEH显著减弱Aβ聚集，降低APP（28%）和BACE（51%）的表达，通过增加BDNF增强突触功能，并恢复ERK磷酸化（<0.05）。抗凋亡作用包括BAX/BCL-2比值降低76%、caspase-3降低47%以及ROS水平降低56%。胆碱能功能通过增加ChAT活性（<0.01）和降低AChE活性（<0.05）得以恢复。PBEH使IL-1β水平降低70%并抑制JNK/p38磷酸化（<0.05）。虽然Com-A增强BDNF且Com-B显示出抗炎作用，但PBEH在多个通路标记物上均表现出活性。总之，这些发现表明PBEH可能通过多通路调节实现神经元保护，为认知增强应用中进一步的机制研究奠定了基础证据。
