Zhang Shuzhi, Guo Jingjing, Suo Shikun, Ju Li, Jiang Zhaoqiang, Dong Pingshuan, Wang Yanli, Dang Yali, Du Laijing
School of Phamacy, Hangzhou Medical College, Hangzhou, China.
Luoyang Key Laboratory of Cardiovascular Science, The First Affiliated Hospital, and College of Clinical Medicine of Henan University of Science and Technology, Luoyang, China.
Front Nutr. 2025 Jan 27;12:1528184. doi: 10.3389/fnut.2025.1528184. eCollection 2025.
Many natural angiotensin-converting enzyme inhibitory (ACEI) peptides have been widely studied. However, their stability is poor in most cases. In this study, peptides were initially digested from broccoli , and absorption was simulated by Caco2 cells transport and then analyzed by Peptideomics and molecular docking. Subsequently, the mechanisms were verified using a high glucose-induced vascular smooth muscle cells (VSMCs) dysfunction model. Results showed that ACEI activity of broccoli crude peptide increased by 70.73 ± 1.42% after digestion. The enzymatic hydrolysates of crude broccoli peptides before and after digestion were detected by HPLC. The digested crude peptides were highly stable (with a stability level > 90%) in the intestine and possessed a strong absorptive potential. Five peptides with high stability and strong permeability were first identified, including HLEVR, LTEVR, LEHGF, HLVNK, and LLDGR, which exhibited high activity with IC values of 3.19 ± 0.23 mM, 17.07 ± 1.37 mM, 0.64 ± 0.02 mM, 0.06 ± 0.01 mM, and 2.81 ± 0.12 mM, respectively. When the VSMCs model was exposed to Ang II, the expressions of PCNA, MMP2, and Bcl2 were increased, while the expression of BAX was inhibited. When the VSMCs was exposed to high glucose (HG), the Ang II concentration significantly increased. This indicates that HG elevated Ang II levels. Finally, five peptides significantly attenuated Ang II-induced VSMCs proliferation and migration by down-regulating AT1R expression and inhibiting ERK and p38 MAPK phosphorylation. Notably, in exploring VSMCs dysfunction on a high glucose-induced model, ACEI peptides resulted in down-regulation of ACE and up-regulation of ACE2 expression. Therefore, it can be further referenced for the functional food against hypertension and cardiovascular diseases.
许多天然的血管紧张素转换酶抑制（ACEI）肽已得到广泛研究。然而，在大多数情况下它们的稳定性较差。在本研究中，肽最初从西兰花中消化得到，通过Caco2细胞转运模拟吸收情况，然后用肽组学和分子对接进行分析。随后，使用高糖诱导的血管平滑肌细胞（VSMC）功能障碍模型验证其作用机制。结果显示，西兰花粗肽消化后的ACEI活性提高了70.73±1.42%。用高效液相色谱法检测消化前后西兰花粗肽的酶解产物。消化后的粗肽在肠道中具有高度稳定性（稳定性水平>90%）且具有很强的吸收潜力。首次鉴定出5种具有高稳定性和强渗透性的肽，包括HLEVR、LTEVR、LEHGF、HLVNK和LLDGR，它们表现出高活性，IC值分别为3.19±0.23 mM、17.07±1.37 mM、0.64±0.02 mM、0.06±0.01 mM和2.81±0.12 mM。当VSMC模型暴露于血管紧张素II（Ang II）时，增殖细胞核抗原（PCNA）、基质金属蛋白酶2（MMP2）和Bcl2的表达增加，而Bax的表达受到抑制。当VSMC暴露于高糖（HG）时，Ang II浓度显著增加。这表明HG升高了Ang II水平。最后，5种肽通过下调血管紧张素II 1型受体（AT1R）表达并抑制细胞外信号调节激酶（ERK）和p38丝裂原活化蛋白激酶（MAPK）磷酸化，显著减弱了Ang II诱导的VSMC增殖和迁移。值得注意的是，在探索高糖诱导模型中的VSMC功能障碍时，ACEI肽导致血管紧张素转换酶（ACE）下调和血管紧张素转换酶2（ACE2）表达上调。因此，它可进一步作为抗高血压和心血管疾病功能性食品的参考。
