Bioavailability and Metabolism of Bioactive Peptide IRW with Angiotensin-Converting Enzyme 2 (ACE2) Upregulatory Activity in Spontaneously Hypertensive Rats.

Peptide IRW is the first food-derived angiotensin-converting enzyme 2 (ACE2) upregulator. This study aimed to investigate the pharmacokinetic characteristics of IRW and identify the metabolites contributing to its antihypertensive activity in spontaneously hypertensive rats (SHRs). Rats were administered 100 mg of IRW/kg of the body weight via an intragastric or intravenous route. The bioavailability (F %) was determined to be 11.7%, and the half-lives were 7.9 ± 0.5 and 28.5 ± 6.8 min for gavage and injection, respectively. Interestingly, significant blood pressure reduction was not observed until 1.5 h post oral administration, or 2 h post injection, indicating that the peptide's metabolites are likely responsible for the blood pressure-lowering activity. Time-course metabolomics revealed a significant increase in the level of kynurenine, a tryptophan metabolite, in blood after IRW administration. Kynurenine increased the level of ACE2 in cells. Oral administration of tryptophan (W), but not dipeptide IR, lowered the blood pressure and upregulated aortic ACE2 in SHRs. Our study supports the key role of tryptophan and its metabolite, kynurenine, in IRW's blood pressure-lowering effects.

Chicken Muscle-Derived ACE2-Upregulating Peptide VVHPKESF Reduces Blood Pressure Associated with the ACE2/Ang (1-7)/MasR Axis in Spontaneously Hypertensive Rats.

SCOPE: This study aims to investigate the antihypertensive effect of four chicken muscle-derived angiotensin (Ang)-converting enzymes (ACE)-regulating peptides: Val-Arg-Pro (VRP, ACE inhibition), Leu-Lys-Tyr and Val-Arg-Tyr (LKY and VRY, ACE inhibition and ACE2 upregulation), and Val-Val-His-Pro-Lys-Glu-Ser-Phe (VVHPKESF [V-F], ACE2 upregulation) in spontaneously hypertensive rats. METHODS AND RESULTS: Rats (12-14 weeks old) are grouped: 1) untreated, 2) VRP, 3) LKY, 4) VRY, and 5) V-F. Blood pressure (BP) is monitored using implantable telemetry technology. Over 18-day oral administration of 15 mg kg-1 body weight (BW) per day, only peptide V-F significantly (p < 0.05) reduces BP, decreases circulating Ang II, and increases ACE2 and Ang (1-7) levels, and enhances aortic expressions of ACE2 and Mas receptor (MasR). Peptide V-F also attenuates vascular inflammation (TNFα, MCP-1, IL-1α, IL-15, and cyclooxygenase 2 [COX2]) and vascular oxidative stress (nitrotyrosine). The gastrointestinal (GI)-degraded fragment of peptide V-F, Val-Val-His-Pro-Lys (VVHPK), is also an ACE2-upregulating peptide. Peptides VRP, LKY, and VRY do not reduce BP, possibly due to low bioavailability or other unknown reasons. CONCLUSIONS: Peptide V-F is the first ACE2-upregulating peptide, purified and fractionated from food proteins based on in vitro ACE2 upregulation, that reduces BP associated with the activation of ACE2/Ang (1-7)/MasR axis; the N-terminal moiety VVHPK may be responsible for the antihypertensive effect of V-F.

Dual cryoprotective and antioxidant effects of young apple polyphenols on myofibrillar protein degradation and gelation properties of bighead carp mince during frozen storage.

Commercial cryoprotectants can delay quality loss in frozen fish mince, but they are associated with a sweet taste and high calorie content. Young apple polyphenols (YAP), extracted from unripe apples, show potential as an alternative cryoprotectant. This study evaluated the cryoprotective effect of YAP at varying levels (0.3%, 0.7%, and 1%) in unwashed bighead carp mince. The changes in sulfhydryl content, carbonyl content, thiobarbituric acid reactive substances, intrinsic fluorescence intensity, and Fourier transform infrared spectrum indicated that YAP retarded oxidation and structural changes in myofibrillar proteins during the first 8 weeks of frozen storage, as well as lipid oxidation, which protected the structure of myofibrillar protein. At higher concentrations (0.7% and 1%), YAP maintained gel properties, gel springiness, and water-holding capacity of the gel prepared from frozen fish mince, potentially through the promotion of cross-linking of myofibrillar proteins. Overall, YAP can be used as a cryoprotectant and antioxidant in fish mince. PRACTICAL APPLICATION: Our research found that young apple polyphenols have the potential to be an alternative to commercial cryoprotectants. Young apple polyphenols may be used as a sugar-free and healthy cryoprotectant for frozen fish mince production in the future.

Angiotensin-Converting Enzyme 2 Activation Is Not a Common Feature of Angiotensin-Converting Enzyme Inhibitory Peptides.

Angiotensin-converting enzyme (ACE) catalyzes the formation of angiotensin II (Ang II), a vasoconstrictor, whereas its homologue ACE2 degrades Ang II into angiotensin (1-7) (Ang (1-7)), a vasodilator. Given the similarities in structure and their interconnected roles in the regulation of cardiovascular system, this study aims to investigate if ACE-inhibitory (ACEi) peptides can also activate ACE2. About 200 potent ACEi peptides were subjected to molecular docking, 20 peptides were selected for cell and in vitro enzymatic activity studies, and 5 peptides were fed orally to spontaneously hypertensive rats at a dose of 15 mg/kg body weight/day for 7 days. Peptides IKW and RIY showed significant antihypertensive activity with activated circulating/aortic ACE2, circulating Ang (1-7), and decreased Ang II levels. IQY reduced blood pressure, increased Ang (1-7) level, but did not affect ACE and ACE2. Peptides MAW and MRW did not affect blood pressure, ACE, and ACE2. Our study showed that ACE2 activation is not a common feature of ACEi peptides.

ACE2 Activation by Tripeptide IRW (Ile-Arg-Trp) Depends on the G Protein-Coupled Receptor 30 Signaling Cascade.

This study aimed to understand how specific cell-bound receptors influence ACE2 activation by IRW. Our results showed that G protein-coupled receptor 30 (GPR30), a 7-transmembrane domain protein, was involved in IRW-mediated ACE2 increase. IRW treatment (50 µM) significantly increased the GPR30 pool levels (3.2 ± 0.5 folds) (p < 0.001). IRW treatment also boosted the consecutive GEF (guanine nucleotide exchange factor) activity (2.2 ± 0.2 folds) (p < 0.001), and GNB1 levels (2.0 ± 0.5 folds) (p < 0.05), associated with the functional subunits of G proteins, in cells. These results were translated in hypertensive animal studies as well (p < 0.05), indicated by an increase in the aortal levels of GPR30 (p < 0.01); further experiments showed an increase in downstream PIP3/PI3K/Akt pathway activation following IRW treatment. The blockade of GPR30 by an antagonist and siRNA in cells abolished the ACE2-activating ability of IRW, as shown by the depleted levels of ACE2 mRNA (p < 0.001), protein levels in whole cells and membrane, angiotensin (1-7) (p < 0.01), and ACE2 promoter HNF1α (p < 0.05). Finally, the GPR30 blockade in ACE2-overexpressing cells using the antagonist (p < 0.01) and siRNA (p < 0.05) significantly depleted the innate cellular pool of ACE2, thus confirming the relationship between the membrane-bound GPR30 and ACE2. Overall, these results showed that the vasodilatory peptide IRW could activate ACE2 via the membrane-bound receptor GPR30.

Novel Hypocholesterolemic Peptides Derived from Silver Carp Muscle: The Modulatory Effects on Enterohepatic Cholesterol Metabolism In Vitro and In Vivo.

This research aimed to investigate the effect of silver carp hydrolysates (SCHs) on hypercholesterolemia and enterohepatic cholesterol metabolism. Results showed that in vitro gastrointestinal digestion products of Alcalase-SCH (GID-Alcalase) exhibited the highest inhibitory activity of cholesterol absorption mainly through downregulating the expression of essential genes related to cholesterol transport in a Caco-2 monolayer. After being absorbed by the Caco-2 monolayer, GID-Alcalase increased the low-density lipoprotein (LDL) uptake in HepG2 cells by enhancing the protein level of the LDL receptor (LDLR). The in vivo experiment showed that long-term intervention of Alcalase-SCH ameliorated hypercholesterolemia in ApoE-/- mice fed with a Western diet (WD). After transepithelial transport, four novel peptides (TKY, LIL, FPK, and IAIM) were identified, and these peptides possessed dual hypocholesterolemic functions including inhibition of cholesterol absorption and promotion of peripheric LDL uptake. Our results indicated for the first time the potential of SCHs as functional food ingredients for the management of hypercholesterolemia.

Peptidomics Study of Plant-Based Meat Analogs as a Source of Bioactive Peptides.

The demand for plant-based meat analogs (PBMA) is on the rise as a strategy to sustain the food protein supply while mitigating environmental change. In addition to supplying essential amino acids and energy, food proteins are known sources of bioactive peptides. Whether protein in PBMA affords similar peptide profiles and bioactivities as real meat remains largely unknown. The purpose of this study was to investigate the gastrointestinal digestion fate of beef and PBMA proteins with a special focus on their potential as precursors of bioactive peptides. Results showed that PBMA protein showed inferior digestibility than that in beef. However, PBMA hydrolysates possessed a comparable amino acid profile to that of beef. A total of 37, 2420 and 2021 peptides were identified in the gastrointestinal digests of beef, Beyond Meat and Impossible Meat, respectively. The astonishingly fewer peptides identified from beef digest is probably due to the near-full digestion of beef proteins. Almost all peptides in Impossible Meat digest were from soy, whereas 81%, 14% and 5% of peptides in Beyond Meat digest were derived from pea, rice and mung proteins, respectively. Peptides in PBMA digests were predicted to exert a wide range of regulatory roles and were shown to have ACE inhibitory, antioxidant and anti-inflammatory activities, supporting the potential of PBMA as a source of bioactive peptides.

Soybean-Derived Tripeptide Leu-Ser-Trp (LSW) Protects Human Vascular Endothelial Cells from TNFα-Induced Oxidative Stress and Inflammation via Modulating TNFα Receptors and SIRT1.

Soybean is a rich source of high-quality proteins and an excellent food source of bioactive peptides. A tripeptide, Leu−Ser−Trp (LSW), was previously identified from soybean as an angiotensin-converting enzyme inhibitory peptide. In the present work, we further studied its antioxidant and anti-inflammatory activities in human vascular endothelial cells (EA.hy926) and elucidated the mechanisms underlying these biological activities. In tumor necrosis factor alpha (TNFα)-stimulated EA.hy926 cells, LSW significantly inhibited oxidative stress, both reduced superoxide and malondialdehyde levels (p < 0.001), owing to its free-radical-scavenging ability. LSW treatment also mitigated the elevated protein expression of vascular adhesion molecule-1 (p < 0.001) and cyclooxygenase 2 (p < 0.01) via inhibition of NF-κB and p38/JNK signaling, respectively. Additionally, LSW also inhibited the endogenous formation of TNFα and attenuated the expression of its two receptors in EA.hy926 cells. Furthermore, LSW upregulated sirtuin-1 level, which partially contributed to its anti-inflammatory activity. These results demonstrate the multiple roles of LSW in ameliorating vascular endothelial oxidative stress and inflammatory responses, which support its uses as a nutraceutical or functional food ingredient for combating endothelial dysfunction and cardiovascular diseases.

Chicken Muscle Protein-Derived Peptide VVHPKESF Reduces TNFα-Induced Inflammation and Oxidative Stress by Suppressing TNFR1 Signaling in Human Vascular Endothelial Cells.

SCOPE: This study aims to investigate the protective effects of four chicken muscle-derived peptides Val-Arg-Pro (VRP), Leu-Lys-Tyr (LKY), Val-Arg-Tyr (VRY), and Val-Val-His-Pro-Lys-Glu-Ser-Phe [VVHPKESF (V-F)] on tumor necrosis factor alpha (TNFα)-induced endothelial inflammation and oxidative stress in human vascular endothelial EA.hy926 cells. METHODS AND RESULTS: Inflammation and oxidative stress are induced in EA.hy926 cells by TNFα (10 ng mL-1 ) treatment for different periods of time. Inflammatory proteins and signaling molecules including inducible nitric oxide synthase, intracellular cell adhesion molecule-1, vascular cell adhesion molecule-1 (VCAM-1), cyclooxygenase 2 (COX2), nuclear factor kappa B (NF-κB), mitogen-activated protein kinases (MAPKs), and TNFα receptor 1 (TNFR1) are measured by qRT-PCR or western blotting; soluble TNFR1 level and nicotinamide adenine dinucleotide phosphate NADPH) oxidase activity are determined by Elisa kits; superoxide is measured by dihydroethidium staining. Only V-F treatment inhibits the expression of VCAM-1 and COX2, via suppressing NF-κB and p38 MAPK signaling, respectively, while reduced oxidative stress via the inhibition of NADPH oxidase activity; V-F treatment attenuates both gene and protein expressions of TNFR1. CONCLUSION: V-F treatment ameliorates TNFα-induced endothelial inflammation and oxidative stress likely via the inhibition of TNFR1 signaling, suggesting its potential as a functional food ingredient or nutraceutical in the prevention and treatment of hypertension and cardiovascular diseases.

Chicken Muscle-Derived ACE2 Upregulating Peptide VVHPKESF Inhibits Angiotensin II-Stimulated Inflammation in Vascular Smooth Muscle Cells via the ACE2/Ang (1-7)/MasR Axis.

This study aimed to evaluate the modulatory effects of four chicken muscle-derived peptides [VRP, LKY, VRY, and VVHPKESF (V-F)] on angiotensin II (Ang II)-induced inflammation in rat vascular smooth muscle A7r5 cells. Only V-F could significantly attenuate Ang II-stimulated inflammation via the inhibition of NF-κB and p38 MAPK signaling, being dependent on the Mas receptor (MasR) not on the Ang II type 1 or type 2 receptor (AT1R or AT2R). V-F accelerated Ang II degradation by enhancing cellular ACE2 activity, which was due to ACE2 upregulation other than a direct ACE2 activation. These findings demonstrated that V-F ameliorated Ang II-induced inflammation in A7r5 cells via the ACE2/Ang (1-7)/MasR axis. Three peptide metabolites of V-F─VHPKESF, PKESF, and SF─were identified but were not considered major contributors to V-F's bioactivity. The regulation of peptide V-F on vascular inflammation supported its functional food or nutraceutical application in the prevention and treatment of hypertension and cardiovascular diseases.

Conventional use and sustainable valorization of spent egg-laying hens as functional foods and biomaterials: A review.

Spent hen are egg-laying hens reaching the end of their laying cycles; billions of spent hens are produced globally each year. Differences in people's attitudes towards spent hen as foods lead to their different fates among countries. While spent hens are consumed as raw or processed meat products in Asian countries such as China, India, Korea, and Thailand, they are treated as a byproduct or waste, not a food product, in the western society; they are instead disposed by burial, incineration, composting (as fertilizers), or rendering into animal feed and pet food, which either create little market value or cause animal welfare and environmental concerns. Despite being a waste, spent hen is a rich source of animal proteins and lipids, which are suitable starting materials for developing valorized products. This review discussed the conventional uses of spent hens, including food, animal feed, pet food, and compost, and the emerging uses, including biomaterials and functional food ingredients. These recent advances enable more sustainable utilization of spent hen, contributing to alternative solutions to its disposal while yielding residual value to the egg industry. Future research will continue to focus on the conversion of spent hen biomass into value-added products.

Chicken muscle hydrolysate reduces blood pressure in spontaneously hypertensive rats, upregulates ACE2, and ameliorates vascular inflammation, fibrosis, and oxidative stress.

Spent hens are egg-laying chicken reaching the end of their egg-laying cycle and are seen as a by-product to the egg industry. A spent hen muscle protein hydrolysate prepared by food-grade thermoase PC10F (SPH-T) has previously shown antihypertensive potential. In the present work, we further investigated its antihypertensive effect and underlying mechanisms in spontaneously hypertensive rats. There are three groups: untreated, low dose (250 mg SPH-T/kg/day body weight), and high dose (1,000 mg SPH-T/kg/day body weight). Oral administration of SPH-T over a period of 20 days reduced systolic blood pressure by 25.7 mm Hg (p < 0.001) and 11.9 mm Hg (p < 0.05), respectively, for the high- and low-dose groups. The high-dose treatment decreased the circulating level of angiotensin II (from 25.0 to 5.7 pg/ml) while increased angiotensin-converting enzyme 2 (ACE2) (from 1.3 to 3.3 IU/ml) and angiotensin (1-7) (from 37.0 to 70.1 pg/ml) significantly (p < 0.05). Furthermore, the high-dose group doubled the aortic expression of ACE2 while reduced the expression of angiotensin (Ang) II type 1 receptor (by 35%). Circulating inflammatory cytokines including tumor necrosis factor alpha and monocyte chemoattractant protein-1 as well as vascular inflammatory proteins including inducible nitric oxide synthase and vascular cell adhesion molecule-1 were attenuated by ∼15%-50% by the treatment; nitrosative stress (35%) and type I collagen synthesis (50%) in the aorta were also attenuated significantly (p < 0.05). Moreover, SPH-T possessed an umami taste (no obvious bitter taste) as analyzed by electronic tongue. PRACTICAL APPLICATION: Hypertension is a global health concern, afflicting more than 20% of adults worldwide. Uncovering the antihypertensive effect of spent hen protein hydrolysate underpinned its functional food nutraceutical applications for the prevention and treatment of hypertension.

Purification of Extracellular Protease from Staphylococcus simulans QB7and Its Ability in Generating Antioxidant and Anti-inflammatory Peptides from Meat Proteins.

Proteases, especially microbial proteases, are widely used in food processing. The purpose of this study was aimed to purify an extracellular protease produced by the strain Staphylococcus simulans QB7 and to evaluate its ability in hydrolyzing meat proteins and generating antioxidant and anti-inflammatory peptides. The optimal conditions for producing the enzyme were as follows: inoculum ratio, 10%; initial pH, 6.5; temperature, 32 °C; incubation time, 36 h; and rotation speed, 160 rpm. The protease had a molecular weight of approximately 47 kDa, possessing the optimal activity at 50 °C, pH 7.0, The protease was stable at pH 4.0-8.0 and 30-60 °C, and the activity was improved by Na+, Mg2+, Ca2+, and Zn2+ ions, whereas it was inhibited by Cu2+, Co2+, Fe3+, Ba2+, Fe2+, ß-M, and ethylene diamine tetraacetic acid disodium salt (EDTA). The protease could effectively hydrolyze meat proteins, and the generated hydrolysate could significantly inhibit tumor necrosis factor-alpha (TNFα)-induced oxidative stress, including superoxide and malondialdehyde levels and inflammation (vascular adhesion molecule-1 [VCAM-1] and cyclooxygenase 2 [COX2)) in human vascular EA.hy926 cells. The present findings support the ability of S. simulans QB7 protease in generating antioxidant and anti-inflammatory peptides during the fermentation of meat products.

Amylase enhances production of low molecular weight collagen peptides from the skin of spent hen, bovine, porcine, and tilapia.

Low molecular weight (LMW) collagen peptides show skin and bone health benefits for human. However, the production of LMW collagen peptides from land vertebrate sources remains challenging due to the presence of advanced glycation end products (AGEs) cross-links. In this study, the effect of α-amylase pre-treatment on proteolytic production of LMW collagen peptides by papain was investigated; spent hen, bovine, porcine, and tilapia skin collagens (HSC, BSC, PSC, and TSC, respectively) were chosen. Results showed that pre-treatment with α-amylase considerably improved the production of LMW peptides (<2 kDa) from HSC (33.79-67.66%), PSC (86.03-90.85%), BSC (6.60-28.78%), and TSC (89.92-90.27%). The HSC presented the highest carbohydrate content and was increased the most in LMW peptides after amylase pretreatment. These results suggested that α-amylase could cleave glycosidic bonds of AGEs between collagen and thus enhance the production of LMW collagen peptides.

Spent Hen Muscle Protein-Derived RAS Regulating Peptides Show Antioxidant Activity in Vascular Cells.

Spent hens are egg-laying hens reaching the end of their egg-laying cycles, being a major byproduct of the egg industry. Recent studies have been focusing on finding new value-added uses for spent hens. We have previously identified four bioactive peptides from spent hen muscle proteins, including three angiotensin-converting enzyme (ACE) inhibitory (ACEi) peptides (VRP, LKY, and VRY), and one ACE2 upregulating (ACE2u) peptide (VVHPKESF (V-F)). In the current study, we further assessed their antioxidant and cytoprotective activities in two vascular cell lines-vascular smooth muscle A7r5 cells (VSMCs) and endothelial EA.hy926 cells (ECs)-upon stimulation by tumor necrosis factor alpha (TNFα) and angiotensin (Ang) II, respectively. The results from our study revealed that all four peptides attenuated oxidative stress in both cells. None of the investigated peptides altered the expression of TNFα receptors in ECs; however, VRY and V-F downregulated Ang II type 1 receptor (AT1R), while V-F upregulated the Mas receptor (MasR) in VSMCs. Further, we found that the antioxidant effects of VRP, LKY, and VRY were likely through acting as direct radical scavengers, while that of V-F was at least partially ascribed to increased endogenous antioxidant enzymes (GPx4 and SOD2) in both cells. Besides, as an ACE2u peptide, V-F exerted antioxidant effect in a MasR-dependent manner, indicating a possible involvement of the upregulated ACE2-MasR axis underlying its antioxidant action. The antioxidant activities of VRP, LKY, VRY, and V-F in vascular cells indicated their multifunctional properties, in addition to their ACEi or ACE2u activity, which supports their potential use as functional food ingredients against hypertension.

Purification and identification of novel ACE inhibitory and ACE2 upregulating peptides from spent hen muscle proteins.

The study explored the use of spent hen, a major egg industry byproduct, as the starting material for preparing antihypertensive peptides. While previous studies were focused mainly on ACE inhibitory (ACEi) peptides, this work also studied peptides with ACE2 upregulating (ACE2u) activity, an emerging target for treating hypertension. Spent hen muscle protein hydrolysate prepared by thermoase (SPH-T) exhibited both ACEi and ACE2u activities. After ultrafiltration and chromatographic fractionation, five potent ACEi peptides, VRP, LKY, VRY, KYKA, and LKYKA, with IC50 values of 0.034-5.77 µg/mL, respectively, and four ACE2u peptides, VKW, VHPKESF, VVHPKESF and VAQWRTKYETDAIQRTEELEEAKKK, which increased ACE2 expression by 0.52-0.84 folds, respectively, were identified; VKW also showed ACEi activity. All peptides, except for VRP, are susceptible to degradation during the simulated gastrointestinal digestion. Our study supports the potential use of spent hens as antihypertensive functional food ingredients and nutraceuticals.

Spent Hen Protein Hydrolysate with Good Gastrointestinal Stability and Permeability in Caco-2 Cells Shows Antihypertensive Activity in SHR.

Spent hens are a major byproduct of the egg industry but are rich in muscle proteins that can be enzymatically transformed into bioactive peptides. The present study aimed to develop a spent hen muscle protein hydrolysate (SPH) with antihypertensive activity. Spent hen muscle proteins were hydrolyzed by nine enzymes, either individually or in combination; 18 SPHs were assessed initially for their in vitro angiotensin-converting enzyme (ACE) inhibitory activity, and three SPHs, prepared by Protex 26L (SPH-26L), pepsin (SPH-P), and thermoase (SPH-T), showed promising activity and peptide yield. These three hydrolysates were further assessed for their angiotensin-converting enzyme 2 (ACE2) upregulating, antioxidant, and anti-inflammatory activities; only SPH-T upregulated ACE2 expression, while all three SPHs showed antioxidant and anti-inflammatory activities. During simulated gastrointestinal digestion, ACE2 upregulating, ACE inhibitory and antioxidant activities of SPH-T were not affected, but those of SPH-26L and SPH-P were reduced. ACE inhibitory activity of gastrointestinal-digested SPH-T was not affected after the permeability study in Caco-2 cells, while ACE2 upregulating, antioxidant and anti-inflammatory activities were improved; nine novel peptides with five-eight amino acid residues were identified from the Caco-2 permeate. Among these three hydrolysates, only SPH-T reduced blood pressure significantly when given orally at a daily dose of 1000 mg/kg body weight to spontaneously hypertensive rats. SPH-T can be developed into a promising functional food ingredient against hypertension, contributing to a more sustainable utilization for spent hens while generating extra revenue for the egg industry.

Regulatory Effects of a Pea-Derived Peptide Leu-Arg-Trp (LRW) on Dysfunction of Rat Aortic Vascular Smooth Muscle Cells against Angiotensin II Stimulation.

Vascular oxidative stress, inflammatory response, and proliferation are crucial mediators of vascular dysfunction which contribute to the pathology of hypertension. A tripeptide, LRW (Leu-Arg-Trp), was characterized from pea protein legumin, and its previously studied isomer IRW (Ile-Arg-Trp) was reported to exhibit antihypertensive activity via activation of angiotensin-converting enzyme 2. The objective of the current study was to explore the effects of LRW on vascular stress in vascular smooth muscle cells (VSMCs) under angiotensin II (Ang II)-induced cellular stress. LRW treatment could decrease Ang II-triggered superoxide production, inflammation, and proliferation in VSMCs. The abovementioned advantageous effects appeared to involve the upregulation of the ACE2-Ang-(1-7)-MasR axis and modulation of the nuclear factor-κB pathway. These findings specified the prospective role of LRW as a functional food ingredient or nutraceutical in the prevention of cardiovascular diseases, particularly hypertension and vascular damage.

Preparation of low-molecular-weight, collagen hydrolysates (peptides): Current progress, challenges, and future perspectives.

Collagen hydrolysates (peptides) derived from food processing byproducts have been used to produce commercially valuable food ingredients due to their potential to trigger certain desirable physiological responses in the body. Low-molecular-weight (LMW) collagen hydrolysates are generally thought to exert better bioactivities than their larger counterparts. However, the preparation of LMW collagen hydrolysates is often impeded by their special structure, cross-linking, and hydroxyproline. This review briefly introduces the motivation of the food industry to prepare LMW collagen hydrolysate from food processing byproducts. We further summarize recent progress on the preparation of LMW collagen hydrolysates and methods to determine the molecular weight. We then discuss the challenges and then provide perspectives on future directions in preparing LMW collagen hydrolysates.

Molecular interactions, bioavailability, and cellular mechanisms of angiotensin-converting enzyme inhibitory peptides.

Food-derived angiotensin-converting enzyme inhibitory (ACEi) peptides have gained substantial interest as potential alternatives to synthetic drugs in the management of hypertension. Peptide size and sequence are two critical factors that determine their potency, bioavailability, and cellular mechanisms. Molecular interaction studies between ACE and ACEi peptides support that potent ACEi peptides are generally composed of hydrophobic, positively charged, and aromatic or cyclic amino acid residues at the third, second, and first position from the C-terminus, respectively. Small peptides containing N-terminal Tyr and/or C-terminal Pro could improve their stability against enterocyte peptidases, thus their bioavailability. Different ACEi peptides can reduce aberrant cellular proliferation, excessive inflammation, and oxidative stress but through different mechanisms. Further understanding the structure-activity-bioavailability relationships will help design novel potent ACEi peptides with improved bioavailability and in vivo efficacy. PRACTICAL APPLICATIONS: ACEi peptides have the potential for uses as functional food ingredients against hypertension.