Home Journals Articles Updates About
CN
image
收藏切换
Study of screening, transport pathway, and vasodilation mechanisms on angiotensin-I converting enzyme inhibitory peptide from Ulva prolifera proteins
收藏切换
PDF
Zhiyong Li1, Yuan He1, Hongyan He1, Caiwei Fu1, Mengru Li1, Aiming Lu1, Dongren Zhang1, Tuanjie Che2, Songdong Shen1, *
Acta Oceanologica Sinica | 2023, 42(11) : 98 - 106
Less
收藏切换
Acta Oceanologica Sinica | 2023, 42(11): 98-106
• Marine Biology •
Study of screening, transport pathway, and vasodilation mechanisms on angiotensin-I converting enzyme inhibitory peptide from Ulva prolifera proteins
Full
Zhiyong Li1, Yuan He1, Hongyan He1, Caiwei Fu1, Mengru Li1, Aiming Lu1, Dongren Zhang1, Tuanjie Che2, Songdong Shen1, *
Affiliations
  • 1 School of Biology & Basic Medical Sciences, Soochow University, Suzhou 215101, China
  • 2 Key Laboratory of Functional Genomic and Molecular Diagnosis of Gansu Province, Lanzhou 730030, China
Published: 2023-11-25 doi: 10.1007/s13131-023-2170-1
Outline
收藏切换

In this study, Ulva prolifera protein was used for preparing angiotensin-I converting enzyme (ACE)-inhibitory peptide via virtual gastrointestinal digestion and in silico screening. Some parameters of the obtained peptide, such as inhibition kinetics, docking mechanism, stability, transport pathway, were explored by Lineweaver-Burk plots, molecular docking, in vitro stimulate gastrointestinal (GI) digestion and Caco-2 cells monolayer model, respectively. Then, a novel anti-ACE peptide LDF (IC50, (1.66 ± 0.34) μmol/L) was screened and synthesized by chemical synthesis. It was a no-competitive inhibitor and its anti-ACE inhibitory effect mainly attributable to four Conventional Hydrogen Bonds and Zn701 interactions. It could keep activity during simulated GI digestion in vitro and was transported by peptide transporter PepT1 and passive-mediated mode. Besides, it could activate Endothelial nitric oxide synthase (eNOS) activity to promote the production of NO and reduce Endothelin-1 (ET-1) secretion induced by Angiotensin II (Ang II) in Human Umbilical Vein Endothelial Cells (HUVECs). Meanwhile, it could promote mice splenocytes proliferation in a concentration-dependent manner. Our study indicated that this peptide was a potential ingredient functioning on vasodilation and enhancing immunity.

Ulva prolifera protein  /  ACE-inhibitory peptide  /  in vitro GI digestion  /  vasodilation mechanism  /  immunomodulation
Zhiyong Li, Yuan He, Hongyan He, Caiwei Fu, Mengru Li, Aiming Lu, Dongren Zhang, Tuanjie Che, Songdong Shen. Study of screening, transport pathway, and vasodilation mechanisms on angiotensin-I converting enzyme inhibitory peptide from Ulva prolifera proteins[J]. Acta Oceanologica Sinica, 2023 , 42 (11) : 98 -106 . DOI: 10.1007/s13131-023-2170-1
Full Text
Less
1 Introduction
Less
Ulva prolifera, an edible green alga, grows on rocks in intertidal zones on mud beaches or on other floating seaweeds on the sea (Tan et al., 1999). This alga contains 9%−14% protein, 2%−3.6% ether extract, 32%−36% ash, and n-3 and n-6 fatty acids 10.4 g/(100 g) and 10.9 g/(100 g) of total fatty acid (Aguilera-Morales et al., 2005). Meanwhile, it’s an important source of protein for human consumptions, attributable to its protein composition, containing eighteen kinds of amino acids, including all essential amino acids and a high protein digestibility (98%) (He et al., 2021). The eutrophication of sea water caused by industrial and domestic sewage provides favorable living conditions for U. prolifera, resulting in the excess production of this alga (Ye et al., 2011). Nearly all the algae are used as low economical-value food, animal feed, fertilizer or even regarded as industrial waste (Li et al., 2016). Thus, increasing the added value of U. prolifera is important in algae’s comprehensive utilization.
Hypertension, a chronic disease, afflicting more than 30% of adults worldwide, is the leading cause of cardiovascular disease and premature death (Mills et al., 2020). ACE is a vital enzyme in blood pressure regulation through both the renin-angiotensin system (RAS) and kallikrein kinnin system (KKS), which catalyzes the conversion of inactive Angiotensin I (Ang I) into a potent vasoconstrictor Ang II to inactivate the vasodilator bradykinin (Raghavan and Kristinsson, 2009). Therefore, screening ACE inhibitor is considered as an effective therapeutic approach for the treatment of hypertension. However, some synthetic ACE inhibitors, such as Captopril, Lisinopril, and Enalapril, may lead to significant side effect like inflammatory response, cough, or renal impairment (Ko et al., 2012). Recently, numerous algae proteins have been utilized to isolate ACE inhibitory peptides, such as Mazzaella japonica protein (Kumagai et al., 2020), Gracilariopsis lemaneiformis protein (Cao et al., 2017) and Grateloupia asiatica protein (Sumikawa et al., 2020). However, until now, no anti-ACE peptide purified form U. prolifera protein has been synthesized and reached clinical trials.
On the other hand, increasingly, various immunomodulatory peptides have been isolated from plant protein, such as wheat gluten protein (Horiguchi et al., 2005), Juglans regia L protein (Mao et al., 2020), and rice protein (Wen et al., 2021). However, only a few studies of immunomodulatory peptide from U. prolifera protein have been reported.
In recent years, in silico approaches have been used to predict and screen various anti-ACE peptides released from food-derived proteins, such as rice bran protein (Udenigwe, 2016), Chlorella vulgaris protein (Xie et al., 2018) and rabbit meat protein (Chen et al., 2022). In contrast to bench experiments, it can reduce the time required for screening peptides from different protein sources and help to discover new and sustainable precursors of known bioactive peptides (Udenigwe, 2014). Meanwhile, the in vivo anti-hypertension effect of peptides is attributable to its stable structure and high absorptivity in the GI tract. Therefore, it is essential to study the stability of bioactive peptides in the GI tract. Simulated GI digestion is a simple, rapid, and inexpensive measure (Lacroix et al., 2017). Differentiated and polarized Caco-2 monolayer is an effective tool to study the transport of peptides in vitro due to their morphology, secretion of marker enzymes and permeability characteristics, which are similar to those of the human small intestine (Ding et al., 2021).
Base on the above considerations, we aimed to screen a safe ACE inhibitory peptide from U. prolifera protein by in silico method. Some pharmacological properties of the selected peptide, such as immunomodulatory activity, GI stability, inhibitory pattern, intestinal tract transport pathway, molecular interactions mechanism of this peptide was also explored. Lastly, the cellular mechanism of the purified peptide on vasodilation was explored on Human Umbilical Vein Endothelial Cells (HUVECs).
2 Materials and methods
Less
2.1 Materials
Protein sequences were downloaded from NCBI (https://c1n.cn/wny80) and they were selected for in silico analysis. ACE (from rabbit lung), N-hippuril-L-histidy-L-leucine (HHL), Ang II, Human ET-1 ELISA Kit was purchased from Sangon Biotech (Shanghai, China). Nitric Oxide Assay Kit was purchased from Beyotime (Shanghai, China). MTT cell proliferation and cytotoxicity assay kits were purchased from Sigma-Aldrich (St. Louis, MO, USA). HUVECs cell line was purchased from SSRCC (Shanghai, China). All other reagents were of analytical reagent grade.
2.2 Evaluation of Ulva prolifera proteins as sources of ACE inhibitory peptides
Fourteen Ulva prolifera protein sequences as the precursors of ACE inhibitory peptides were evaluated using the BIOPEP-UWM database (https://c1n.cn/yDaOA). The parameters of proteins, such as name, amino acid number, molecular weight (MW) were shown in Table 1. The frequency (A) of anti-ACE peptides was calculated using the following formula:
$ A = a/N, $
where a was the number of peptides with anti-ACE activity in a given protein sequence and N was the total number of amino acid residues of protein.
2.3 In silico digestion and screening
All the proteins were virtually GI digested by complex enzymes (pepsin and trypsin) in PeptideCutter (expasy.org). Biological activity potential was evaluated at PeptideRanker (ucd.ie). Water solubility was calculated at Innovagen AB (Antibodies, proteins, and peptides). Toxicity was explored at ToxinPred (osdd.net). Human intestinal absorption (HIA) was tested at admetSAR (ecust.edu.cn). Finally, the affinities of these peptides against the ACE were evaluated by molecular docking in Discovery Studio 2020 software (DS 2020, Accelrys, San Diego, CA, USA).
2.4 Peptide synthesis and activity validation
The selected biopeptides were synthesized by China Peptides Co., Ltd. (Suzhou, China) and the purity of the peptide was ≥98%. The ACE inhibitory rate was explored according to previous reported (Pan et al., 2016). The IC50 value was defined as the inhibitor concentration inhibiting 50% activity of ACE.
2.5 Inhibition kinetics of the synthetic peptide.
The inhibition pattern of the synthetic peptides was explored by Lineweaver-Burk plots. The concentrations of the ACE substate (HHL) were set as 1 mol/L, 2.5 mol/L and 5 mol/L. Peptide concentrations were set as 0 μmol/L, 10 μmol/L and 100 μmol/L. Vmax and Km were measured as the y-axis and x-axis intercepts of the primary plot, respectively.
2.6 Stability of the synthetic peptides against GI proteases
Peptides solution was adjusted to pH 2.0 (using 1 mol/L HCl). Pepsin (1% E/S (enzyme/substrate), w/w) was added to the solution and the mixture was incubated at 37℃ for 2 h. Then, half of the juice was terminated by boiling water and the other half was adjusted to pH 7.5, and followed digested by trypsin (1% E/S, w/w) for another 2 h at 37℃. Then, the reaction was also terminated by boiling water and all the peptide solutions were adjusted to pH 7.0. Subsequently, both solvents were centrifuged at 12 000 r/min for 5 min. The supernatants were analyzed using Q-Exactive mass spectrometer. Besides, the ACE inhibitory activity of the processed peptide solutions was determined at the same concentration (5 μmol/L).
2.7 Molecular docking
ACE crystal structure (ID: 1O8A) was optimized in DS 2020, removing water molecules and ligand, adding hydrogen and then was set as a receptor. The three-dimensional structure of the selected peptide was constructed, followed and carried for energy minimization optimization. Then, followed a docking program between the selected peptide and ACE special binding sites (coordinates x, 40.79; y, 33.61; and z, 43.38) was carried by C docking. Lisinopril was selected as a control candidate. The other parameters were set as default values except for special instructions.
2.8 Proliferative effects of peptides on splenocyte
Preparation of mice splenocytes refers to our previous report (Li et al., 2020). Splenocyte proliferation assay was conducted by MTT Cell Proliferation and Cytotoxicity Assay Kit (Beyotime, Shanghai, China) and the cell viability rate was calculated as follows:
$\begin{split} {\mathrm{Cell}} \; {\mathrm{viability}}\; (\%) =& ({\mathrm{Optical}} \; {\mathrm{density}} \; {\mathrm{of}}\; {\mathrm{test}}\; {\mathrm{well}} - \\ &\rm {\mathrm{optical}} \; {\mathrm{density}} \; {\mathrm{of}} \; {\mathrm{spontaneous}}) \times \\ &\rm 100/({\mathrm{Optical}} \; {\mathrm{density}} \; {\mathrm{of}} \; {\mathrm{control}} -\\ & \rm {\mathrm{Optical}} \; {\mathrm{density}} \; {\mathrm{of}} \; {\mathrm{spontaneous}}). \end{split}$
2.9 Effect of the selected peptide on HUVECs
HUVECs were cultured in Ham’s F-12K (0.1 mg/mL Heparin, 0.03−0.05 mg/mL ECGs, 10% FBS, 100 U/mL 1% penicillin and 100 U/mL streptomycin) at 37℃ and 5% CO2. Peptides with gradient concentrations (0 μmol/L, 25 μmol/L, 50 μmol/L and 100 μmol/L) were used to culture HUVECs.
Intracellular NO content determination: 100 μL of this cell (5 × 105 cells/mL) was seeded in 96-well plates and cultured by peptides for 12 h, before adding 3-amino, 4-aminomethyl-2', 7'-difluorescein, diacetate (DAF-FM DA) (20 µmol/L) for another 30 min. Then, the intracellular fluorescence intensity of HUVECs was explored by Olympus IX81 fluorescent microscope (Olympus, Tokyo, Japan). The excitation and emission wavelength were 495 nm and 515 nm, respectively.
Extracellular NO content determination: HUVECs were cultured by peptides in the same condition as above and the NO content in medium was explored according to Nitric Oxide Synthase Assay Kit (Beyotime, Shanghai, China). The extracellular UV optical density was explored by microplate reader (Varioskan LUX, Thermo Scientific, USA) at 540 nm.
Intracellular Endothelial nitric oxide synthase (eNOS) activity determination: HUVECs were cultured by peptides in the same condition as above and the eNOS activity was explored according to Nitric oxide synthase test kit (Beyotime, Shanghai, China).
ET-1 secretion determination: HUVECs were cultured in 96-well plates for 12 h with the same condition as above, before adding Ang II (100 nmol/L, final concentration) for another 6 h. Finally, the content of ET-1 in processed cells was determined according to Human ET-1 ELISA Kit (Sangon Biotech, Shanghai, China).
2.10 Statistical analysis
Data were analyzed by one-way analysis of variance (ANOVA), and then Dunnett multiple tests was performed using GraphPad Prism Version 9 (San Diego, CA, USA). Values (mean ± SD) that do not share a common superscript lowercase letter within a column differ significantly (p < 0.05) (n = 3).
3 Results and discussion
Less
3.1 Screening of ACE inhibitory peptides
A previous study has confirmed that some peptide fragments present in U. prolifera protein could inhibit the activity of ACE (Pan et al., 2016). Thus, fourteen proteins from the algae were used as precursors to produce a novel anti-ACE peptide by in silico analysis (Table 1). Parameter A represents the frequency of anti-ACE peptides present in a given protein sequence. Various proteins with a given A value higher than 0.3 have been published, such as quinoa proteins (0.345 1 to 0.420 8) (Guo et al., 2020), rabbit meat protein (0.327 5 to 0.866 6) (Chen et al., 2022) and yak milk casein (0.441 2 to 0.655 5) (Lin et al., 2018). Therefore, to sum up, the higher values of parameter A, the more potential anti-ACE peptides in a given protein sequence (Iwaniak et al., 2020). In this study, the A value of the precursor protein was 0.2517, lower than that of the above-mentioned studies. Thus, we speculated that the high A value was not the determining factor in producing anti-ACE peptides.
In recent years, virtual GI digestion as cheap and efficient technology to produce peptide, has been more and more prominent in anti-ACE peptides production (Chen et al., 2022). Some pharmacological properties, such as the biological activity potential, toxicity, solubility, absorptivity and penetration, should be considered in the screening of novel anti-ACE peptides (Ferreira and Andricopulo, 2019). In addition, small peptides usually have higher anti-ACE activity (Pan et al., 2016). Therefore, peptides smaller than pentapeptide were collected and a total of 654 peptides were obtained after in silico proteolysis (Table S1). Among of them, twelve peptides (Peptide ranker >0.8) exhibited better ADMET characteristics (Table 2). However, among of these peptides, only LDF could dock with ACE. Subsequently, this peptide was synthesized and acquired an IC50 of (1.66 ± 0.34) μmol/L in vitro (Fig. 1). Compared to the selected LDF, various othered reported anti-ACE peptides with similar sequence, such as LDY (IC50, 6.10 μmol/L) (Furuta et al., 2016), DFG (IC50, 44.7 μmol/L) (Balti et al., 2010), CDF (IC50, 192.17 μmol/L) (Chen et al., 2022), VDF (IC50, 6.59 μmol/L) and LPF (IC50, 10.59 μmol/L) (Majumder and Wu, 2009). Thus, we speculated that L (Leu) in N-terminal residues or F (Phe, aromatic amino acids) in C-terminal residues may play a key role in the anti-ACE activity of peptides. It was similar to the previous report that aromatic or cyclic amino acids (F, Phe) were the most common C-terminal residues of highly active ACE inhibitors (Xie et al., 2018).
3.2 Inhibitory kinetics of peptides on ACE
Anti-ACE peptides were reported to play three patterns, including competitive, non-competitive, and mixed-competitive to bind with ACE (Maeno et al., 1996; Xie et al., 2018). In this study, as peptide concentration increased (0−100 μmol/L), Km remained constant while Vmax decreased, which was indicative of non-competitive inhibition (Fig. 2). Similarly, for instance, the peptide CDF from rabbit meat protein was a non-competitive inhibitor. Both of them interacted with free protease and/or protease-substrate complex with similar affinity to inhibit the activity of ACE (Sangsawad et al., 2018).
3.3 Stability of peptide against pepsin, pepsin-trypsin.
In order to determine the stability of the synthetic peptides during the incubation of GI proteases, a simulated GI digestion study was performed in vitro. As shown in Fig. 3, some of the peptides were degraded by pepsin and no noteworthy changes were found after incubation with trypsin. Fortunately, there was no significant change in ACE inhibitory activity of the processed peptides (Fig. 3d). Anti-ACE peptides processed by GI proteases showed different effects: an increase in the activity of the WMY peptide, no change in the IY peptide, and a decrease or even loss in the activity of other synthesized peptides (Xu et al., 2021). It was probable that the small peptide fragments from the GI digestion process have an inhibitory function on ACE.
3.4 Transepithelial transport of peptides across the Caco-2 cell monolayer
Caco-2 cell monolayers transport system is an effective model to study the absorption of bioactive peptides (Hidalgo et al., 1989). The basal surface (B side) of Caco-2 cell model is equivalent to the inner wall of the intestine and has an efflux function. In this study, the result indicated that transport rate of peptide LDF exhibited a time-dependent relationship and up to a maximum value of 6.47% at 90 min (Fig. 4a). Similarly, the transport time was consistent with the anti-ACE peptide LVLPGE (Pei et al., 2021). To determine which pathways were involved in the peptide transport, the effects of different modulators were investigated. Wortmannin, a transcytosis inhibitor, had no effect on the transport of LDF, suggesting that endocytosis was not the major transport mode. Gly-Pro was a suitable substrate for peptide transporter of PepT1 and could combine with the majority of PepT1 in the intestinal model. Compared to the control group, we found that the Papp of Gly-Pro treated group was decreased significantly (P < 0.05) (Fig. 4b). Therefore, we speculated that LDF was transported by PepT1. Meanwhile, Cytochalasin D was a disruptor of tight junction (TJ), and could significantly increase the transport of LDF (P < 0.05), suggesting that TJ was involved in the transport process (Fig. 4b). Therefore, the passive paracellular transport via TJ was involved in the transport process of LDF. A large number of water-soluble short peptides were transported by paracellular pathway, such as LVLPGE (Pei et al., 2021), KYIPIQ (Lin et al., 2020), IQW and LKP (Xu et al., 2017). In this study, LDF was consistent with the above reported peptides. Meanwhile, the monolayers of Caco-2 cells were tighter than that of human intestinal epithelial cells, which was not conducive to peptide transport (Conradi et al., 1993). In contrast, human intestine contains more carrier-mediated transports and a larger effective absorbing area (Ding et al., 2021). Therefore, the transport of LDF in vivo may be superior to that of the Caco-2 cells model.
3.5 Molecular docking
Molecular docking is a key tool in exploring the binding mechanism of inhibitor and ACE (Chen et al., 2022). As shown in Table 3, there were four types of bonds between LDF and ACE, including Electrostatic interaction, Conventional Hydrogen Bond, Carbon Hydrogen Bond, and Alkyl. The most stabilized pose of LDF was mainly attributable to the four conditional Hydrogen bonds, which docked onto the key sites, such as HIS353 in S1 pocket and TYR520 in S2 pocket, respectively. Meanwhile, some key sites in S2 pocket, such as HIS353, HIS513, and LYS511 were linked by Electrostatic interaction and Carbon Hydrogen Bond, respectively. In contrast, there were four types of bonds between Lisinopril and ACE, including Pi-Cation, Attractive Charge, Carbon Hydrogen Bond, Conventional Hydrogen Bond. Notably, the Conventional Hydrogen Bond number of Lisinopril was higher than LDF, which may contribute to the lower IC50 of Lisinopril (IC50 = 1.1 ng/mL) in vitro (Fig. 5). In addition, zinc is an important catalytic component of ACE (Natesh et al., 2003). Paradoxically, LDF as a non-competitive inhibitor can bind with the active site (Zn701) of ACE. Pervious study indicated that some non-competitive anti-ACE peptides, such as PFPGPIPN (Lin et al., 2017), TTW and VHW (Xie et al., 2018) can also bind with the active site of ACE during the molecular docking process. We speculated that the dock result is related to the evaluation mechanism of docking software, in which all the potential dock poses of peptide with ACE will be exhibited.
3.6 Effect of peptide on mice splenocytes.
A large body of evidence shows that bioactive peptides regulate immunity by enhancing lymphocyte proliferation and antibody synthesis (Yang et al., 2009). As shown in Fig. 6, the proliferative effect of LDF on mice splenocytes exhibited concentration-dependence. Some peptides with pro-proliferative activity usually contain a high proportion of hydrophobic amino acids, such as FFSDL (Sütas et al., 1996), FLPFNQL (Liu et al., 2020), RGPPP (Yang et al., 2020) and NGLAP (Hou et al., 2012). Thus, the hydrophobic amino acids Leu (L) and Phe (F) may play a key role in the proliferative effects of LDF. Meanwhile, immune cells play a major role in blood pressure control and have a critical impact as a therapeutic target for hypertension (Liu et al., 2017). The in vivo immunity and blood pressure regulation by LDF in mice need to be further studied.
3.7 Effect of this peptide on eNOS activity, NO and EF-1 secretion in HUVECs
HUVECs line the internal surface of blood vessels, and represent a good model to evaluate the molecular mechanisms involved in blood pressure regulation (García-Tejedor et al., 2015). In this study, In vitro HUVECs incubation was performed to explore the vasodilation mechanism of LDF. As shown in Fig. 7a, LDF (100 μmol/L) could promote the secretion of NO significantly (p < 0.05) by activating the activity of eNOS (P < 0.05). Similarly, the green fluorescence value indicated that, the intracellular NO content increased significantly under the effect of peptides (Fig. 7c). Bradykinin, a peptide fragment containing nine amino acids, acted to promote the production of NO by endothelial cells and it could be degraded by ACE (Fukuda et al., 2009). Hence, we speculated that the anti-ACE effect of LDF reduced the breakdown of bradykinin to promote the activation of eNOS and NO generation. In HUVECs, eNOS converts the L-arginine into L-citrulline and Endothelial NO and it is tightly regulated by co- and post-translational lipid modifications, and phosphorylation on multiple residues (Robb et al., 2004). eNOS-derived NO, an endogenous vasodilatory gas, functioned on vasorelaxation via the activation of vascular smooth muscle soluble guanylate cyclase (sGC) and maintained an anti-proliferative and anti-apoptotic environment in the vessel’s wall (Zabel et al., 1998; Sessa, 2004). However, which signaling pathway play vital role in mediating LDF-stimulate eNOS activation and NO production needs to be further studied. ET-1, an endothelium-derived peptide, was one of the most potent vasoconstrictors (Miyauchi et al., 1990). Ang II could promote the secretion of ET-1 in a concentration-dependent manner though the upregulation of the ET-1 mRNA and its effects were prevented by specific protease inhibitors (Imai et al., 1992). Pervious study indicated vasoconstriction to ET-1 and thromboxane is augmented in the face of eNOS deficiency (Lamping and Faraci, 2003). Similarly, LDF could induce eNOS activation and prevent the Ang II-induced overexpression of ET-1 (P < 0.05) (Fig. 7b). These results suggested the potential anti-hypertension effect of LDF in vivo. In addition, ET-1 could induce NAD(P)H oxidase subunit gp91phox mRNA expression to enhance superoxide anions generation in HUVECs (Duerrschmidt et al., 2000). Therefore, LDF could alleviate vascular endothelial oxidative damage mediated by ET-1.
4 Conclusions
Less
In this study, a novel anti-ACE peptide LDF (IC50, (1.66 ± 0.34) μmol/L) was screened from Ulva prolifera protein. It was a no-competitive inhibitor and its anti-ACE inhibitory effect mainly was attributable to four Conventional Hydrogen Bonds and Zn701 interactions. It could keep activity after simulated GI digestion in vitro and was transported by peptide transporter PepT1 and passive-mediated mode through Caco-2 cells monolayers. Besides, in HUVECs, LDF could activate eNOS activity to promote the production of NO and reduce ET-1 secretion induced by Ang II. In addition, LDF could promote mice splenocytes proliferation in a concentration-dependent manner. Our study indicated that LDF was a potential drug ingredient to relax blood vessels and enhance immunity.
References
Less
Aguilera-Morales M, Casas-Valdez M, Carrillo-Domı́nguez S, et al. 2005. Chemical composition and microbiological assays of marine algae Enteromorpha spp. as a potential food source. Journal of Food Composition and Analysis, 18(1): 79–88, doi: 10.1016/j.jfca.2003.12.012
Balti R, Nedjar-Arroume N, Adjé E Y, et al. 2010. Analysis of novel angiotensin I-converting enzyme inhibitory peptides from enzymatic hydrolysates of cuttlefish (Sepia officinalis) muscle proteins. Journal of Agricultural and Food Chemistry, 58(6): 3840–3846, doi: 10.1021/jf904300q
Cao Dequn, Lv Xiaojing, Xu Xiaoting, et al. 2017. Purification and identification of a novel ACE inhibitory peptide from marine alga Gracilariopsis lemaneiformis protein hydrolysate. European Food Research and Technology, 243(10): 1829–1837, doi: 10.1007/s00217-017-2886-2
Chen Junbo, Yu Xiaodong, Chen Qianzi, et al. 2022. Screening and mechanisms of novel angiotensin-I-converting enzyme inhibitory peptides from rabbit meat proteins: A combined in silico and in vitro study. Food Chemistry, 370: 131070, doi: 10.1016/j.foodchem.2021.131070
Conradi R A, Wilkinson K F, Rush B D, et al. 1993. In vitro/in vivo models for peptide oral absorption: Comparison of Caco-2 cell permeability with rat intestinal absorption of renin inhibitory peptides. Pharmaceutical Research, 10(12): 1790–1792, doi: 10.1023/A:1018990602102
Ding Xiaomeng, Hu Xiaoyi, Chen Yi, et al. 2021. Differentiated Caco-2 cell models in food-intestine interaction study: Current applications and future trends. Trends in Food Science & Technology, 107: 455–465, doi: 10.1016/j.jpgs.2020.11.015
Duerrschmidt N, Wippich N, Goettsch W, et al. 2000. Endothelin-1 induces NAD(P)H oxidase in human endothelial cells. Biochemical and Biophysical Research Communications, 269(3): 713–717, doi: 10.1006/bbrc.2000.2354
Ferreira L L G, Andricopulo A D. 2019. ADMET modeling approaches in drug discovery. Drug Discovery Today, 24(5): 1157–1165, doi: 10.1016/j.drudis.2019.03.015
Fukuda D, Enomoto S, Nagai R, et al. 2009. Inhibition of renin–angiotensin system attenuates periadventitial inflammation and reduces atherosclerotic lesion formation. Biomedicine & Pharmacotherapy, 63(10): 754–761, doi: 10.1016/j.biopha.2009.02.006
Furuta T, Miyabe Y, Yasui H, et al. 2016. Angiotensin I converting enzyme inhibitory peptides derived from phycobiliproteins of dulse Palmaria palmata. Marine Drugs, 14(2): 32, doi: 10.3390/md14020032
García-Tejedor A, Gimeno-Alcañíz J V, Tavárez S, et al. 2015. An antihypertensive lactoferrin hydrolysate inhibits angiotensin I-converting enzyme, modifies expression of hypertension-related genes and enhances nitric oxide production in cultured human endothelial cells. Journal of Functional Foods, 12: 45–54, doi: 10.1016/j.jff.2014.11.002
Guo Huimin, Richel A, Hao Yuqiong, et al. 2020. Novel dipeptidyl peptidase-IV and angiotensin-I-converting enzyme inhibitory peptides released from quinoa protein by in silico proteolysis. Food Science & Nutrition, 8(3): 1415–1422, doi: 10.1002/fsn3.1423
He Yuan, Shen Songdong, Yu Dachun, et al. 2021. The Ulva prolifera genome reveals the mechanism of green tides. Journal of Oceanology and Limnology, 39(4): 1458–1470, doi: 10.1007/s00343-020-0212-5
Hidalgo I J, Raub T J, Borchardt R T. 1989. Characterization of the human colon carcinoma cell line (Caco-2) as a model system for intestinal epithelial permeability. Gastroenterology, 96(3): 736–749, doi: 10.1016/0016-5085(89)90897-4
Horiguchi N, Horiguchi H, Suzuki Y. 2005. Effect of wheat gluten hydrolysate on the immune system in healthy human subjects. Bioscience, Biotechnology, and Biochemistry, 69(12): 2445–2449, doi: 10.1271/bbb.69.2445
Hou Hu, Fan Yan, Li Bafang, et al. 2012. Purification and identification of immunomodulating peptides from enzymatic hydrolysates of Alaska pollock frame. Food Chemistry, 134(2): 821–828, doi: 10.1016/j.foodchem.2012.02.186
Imai T, Hirata Y, Emori T, et al. 1992. Induction of endothelin-1 gene by angiotensin and vasopressin in endothelial cells. Hypertension, 19(6_pt_2): 753–757, doi: 10.1161/01.HYP.19.6.753
Iwaniak A, Minkiewicz P, Pliszka M, et al. 2020. Characteristics of biopeptides released in silico from collagens using quantitative parameters. Foods, 9(7): 965, doi: 10.3390/foods9070965
Ko S C, Kang N, Kim E A, et al. 2012. A novel angiotensin I-converting enzyme (ACE) inhibitory peptide from a marine Chlorella ellipsoidea and its antihypertensive effect in spontaneously hypertensive rats. Process Biochemistry, 47(12): 2005–2011, doi: 10.1016/j.procbio.2012.07.015
Kumagai Y, Kitade Y, Kobayashi M, et al. 2020. Identification of ACE inhibitory peptides from red alga Mazzaella japonica. European Food Research and Technology, 246(11): 2225–2231, doi: 10.1007/s00217-020-03567-z
Lacroix I M E, Chen Xiumin, Kitts D D, et al. 2017. Investigation into the bioavailability of milk protein-derived peptides with dipeptidyl-peptidase IV inhibitory activity using Caco-2 cell monolayers. Food & Function, 8(2): 701–709, doi: 10.1039/C6FO01411A
Lamping K, Faraci F. 2003. Enhanced vasoconstrictor responses in eNOS deficient mice. Nitric Oxide, 8(4): 207–213, doi: 10.1016/S1089-8603(03)00028-4
Li Hongmei, Zhang Yongyu, Han Xiurong, et al. 2016. Growth responses of Ulva prolifera to inorganic and organic nutrients: Implications for macroalgal blooms in the southern Yellow Sea, China. Scientific Reports, 6(1): 26498, doi: 10.1038/srep26498
Li Zhiyong, Zhao Shan, Xin Xiangdong, et al. 2020. Purification, identification and functional analysis of a novel immunomodulatory peptide from silkworm pupa protein. International Journal of Peptide Research and Therapeutics, 26(1): 243–249, doi: 10.1007/s10989-019-09832-4
Lin Kai, Ma Zhao, Ramachandran M, et al. 2020. ACE inhibitory peptide KYIPIQ derived from yak milk casein induces nitric oxide production in HUVECs and diffuses via a transcellular mechanism in Caco-2 monolayers. Process Biochemistry, 99: 103–111, doi: 10.1016/j.procbio.2020.08.031
Lin Kai, Zhang Lanwei, Han Xue, et al. 2017. Novel angiotensin I-converting enzyme inhibitory peptides from protease hydrolysates of Qula casein: Quantitative structure-activity relationship modeling and molecular docking study. Journal of Functional Foods, 32: 266–277, doi: 10.1016/j.jff.2017.03.008
Lin Kai, Zhang Lanwei, Han Xue, et al. 2018. Yak milk casein as potential precursor of angiotensin I-converting enzyme inhibitory peptides based on in silico proteolysis. Food Chemistry, 254: 340–347, doi: 10.1016/j.foodchem.2018.02.051
Liu Ping, Liao Wang, Qi Xingpu, et al. 2020. Identification of immunomodulatory peptides from zein hydrolysates. European Food Research and Technology, 246(5): 931–937, doi: 10.1007/s00217-020-03450-x
Liu Yunmeng, Rafferty T M, Rhee S W, et al. 2017. CD8+ T cells stimulate Na-Cl co-transporter NCC in distal convoluted tubules leading to salt-sensitive hypertension. Nature Communications, 8(1): 14037, doi: 10.1038/ncomms14037
Maeno M, Yamamoto N, Takano T. 1996. Identification of an antihypertensive peptide from casein hydrolysate produced by a proteinase from Lactobacillus helveticus CP790. Journal of Dairy Science, 79(8): 1316–1321, doi: 10.3168/jds.S0022-0302(96)76487-1
Majumder K, Wu Jianping. 2009. Angiotensin I converting enzyme inhibitory peptides from simulated in vitro gastrointestinal digestion of cooked eggs. Journal of Agricultural and Food Chemistry, 57(2): 471–477, doi: 10.1021/jf8028557
Mao Ruixue, Wu Lan, Zhu Na, et al. 2020. Immunomodulatory effects of walnut (Juglans regia L. ) oligopeptides on innate and adaptive immune responses in mice. Journal of Functional Foods, 73: 104068, doi: 10.1016/j.jff.2020.104068
Mills K T, Stefanescu A, He Jiang. 2020. The global epidemiology of hypertension. Nature Reviews Nephrology, 16(4): 223–237, doi: 10.1038/s41581-019-0244-2
Miyauchi T, Tomobe Y, Shiba R, et al. 1990. Involvement of endothelin in the regulation of human vascular tonus. Potent vasoconstrictor effect and existence in endothelial cells. Circulation, 81(6): 1874–1880, doi: 10.1161/01.CIR.81.6.1874
Natesh R, Schwager S L U, Sturrock E D, et al. 2003. Crystal structure of the human angiotensin-converting enzyme–lisinopril complex. Nature, 421(6922): 551–554, doi: 10.1038/nature01370
Saikun Pan, Shujun Wang , Lingling Jing , Dongrui Yao, et al. 2016. Purification and characterisation of a novel angiotensin-I converting enzyme (ACE)-inhibitory peptide derived from the enzymatic hydrolysate of Enteromorpha clathrata protein. Food Chemistry, 211: 423–430, doi: 10.1016/j.foodchem.2016.05.087
Pei Jingyan, Hua Ying, Zhou Tingyi, et al. 2021. Transport, in vivo antihypertensive effect, and pharmacokinetics of an Angiotensin-Converting Enzyme (ACE) inhibitory peptide LVLPGE. Journal of Agricultural and Food Chemistry, 69(7): 2149–2156, doi: 10.1021/acs.jafc.0c07048
Raghavan S, Kristinsson H G. 2009. ACE-inhibitory activity of tilapia protein hydrolysates. Food Chemistry, 117(4): 582–588, doi: 10.1016/j.foodchem.2009.04.058
Robb G B, Carson A R, Tai S C, et al. 2004. Post-transcriptional regulation of endothelial nitric-oxide synthase by an overlapping antisense mRNA transcript. Journal of Biological Chemistry, 279(36): 37982–37996, doi: 10.1074/jbc.M400271200
Sütas Y, Soppi E, Korhonen H, et al. 1996. Suppression of lymphocyte proliferation in vitro by bovine caseins hydrolyzed with Lactobacillus casei GG–derived enzymes. Journal of Allergy and Clinical Immunology, 98(1): 216–224, doi: 10.1016/S0091-6749(96)70245-2
Sangsawad P, Choowongkomon K, Kitts D D, et al. 2018. Transepithelial transport and structural changes of chicken angiotensin I-converting enzyme (ACE) inhibitory peptides through Caco-2 cell monolayers. Journal of Functional Foods, 45: 401–408, doi: 10.1016/j.jff.2018.04.020
Sessa W C. 2004. eNOS at a glance. Journal of Cell Science, 117(12): 2427–2429, doi: 10.1242/jcs.01165
Sumikawa K, Takei K, Kumagai Y, et al. 2020. In silico analysis of ACE inhibitory peptides from chloroplast proteins of red alga Grateloupia asiatica. Marine Biotechnology, 22(3): 391–402, doi: 10.1007/s10126-020-09959-2
Tan I H, Blomster J, Hansen G, et al. 1999. Molecular phylogenetic evidence for a reversible morphogenetic switch controlling the gross morphology of two common genera of green seaweeds, Ulva and Enteromorpha. Molecular Biology and Evolution, 16(8): 1011–1018, doi: 10.1093/oxfordjournals.molbev.a026190
Udenigwe C C. 2014. Bioinformatics approaches, prospects and challenges of food bioactive peptide research. Trends in Food Science & Technology, 36(2): 137–143, doi: 10.1016/j.jpgs.2014.02.004
Udenigwe C C. 2016. Towards rice bran protein utilization: In silico insight on the role of oryzacystatins in biologically-active peptide production. Food Chemistry, 191: 135–138, doi: 10.1016/j.foodchem.2015.01.043
Wen Li, Huang Lu, Li Yiwei, et al. 2021. New peptides with immunomodulatory activity identified from rice proteins through peptidomic and in silico analysis. Food Chemistry, 364: 130357, doi: 10.1016/j.foodchem.2021.130357
Xie Jingli, Chen Xujun, Wu Junjie, et al. 2018. Antihypertensive effects, molecular docking study, and isothermal titration calorimetry assay of angiotensin I-converting enzyme inhibitory peptides from Chlorella vulgaris. Journal of Agricultural and Food Chemistry, 66(6): 1359–1368, doi: 10.1021/acs.jafc.7b04294
Xu Qingbiao, Fan Hongbing, Yu Wenlin, et al. 2017. Transport study of egg-derived antihypertensive peptides (LKP and IQW) using Caco-2 and HT29 coculture monolayers. Journal of Agricultural and Food Chemistry, 65(34): 7406–7414, doi: 10.1021/acs.jafc.7b02176
Xu Zhenqiu, Wu Changping, Sun-Waterhouse D, et al. 2021. Identification of post-digestion angiotensin-I converting enzyme (ACE) inhibitory peptides from soybean protein isolate: Their production conditions and in silico molecular docking with ACE. Food Chemistry, 345: 128855, doi: 10.1016/j.foodchem.2020.128855
Yang Qian, Cai Xixi, Huang Muchen, et al. 2020. A specific peptide with immunomodulatory activity from Pseudostellaria heterophylla and the action mechanism. Journal of Functional Foods, 68: 103887, doi: 10.1016/j.jff.2020.103887
Yang Ruiyue, Zhang Zhaofeng, Pei Xinrong, et al. 2009. Immunomodulatory effects of marine oligopeptide preparation from Chum Salmon (Oncorhynchus keta) in mice. Food Chemistry, 113(2): 464–470, doi: 10.1016/j.foodchem.2008.07.086
Ye Naihao, Zhang Xiaowen, Mao Yuze, et al. 2011. ‘Green tides’ are overwhelming the coastline of our blue planet: taking the world’s largest example. Ecological Research, 26(3): 477–485, doi: 10.1007/s11284-011-0821-8
Zabel U, Weeger M, La M, et al. 1998. Human soluble guanylate cyclase: functional expression and revised isoenzyme family. Biochemical Journal, 335(1): 51–57, doi: 10.1042/bj3350051
Appendix
Less
Year 2023 volume 42 Issue 11
PDF
77
42
Cite this Article
BibTeX
Article Info
doi: 10.1007/s13131-023-2170-1
  • Receive Date：2022-07-29
  • Online Date：2025-11-22
  • Published：2023-11-25
Article Data
Affiliations
History
  • Received：2022-07-29
  • Accepted：2023-01-11
Affiliations
    1 School of Biology & Basic Medical Sciences, Soochow University, Suzhou 215101, China
    2 Key Laboratory of Functional Genomic and Molecular Diagnosis of Gansu Province, Lanzhou 730030, China

Corresponding:

* Songdong Shen Email:
References
Share
https://castjournals.cast.org.cn/joweb/aos/EN/10.1007/s13131-023-2170-1
Share to
QR

Scan QR to access full text

Cite this article
BibTeX
Citations
表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
关闭全屏
  • BibTeX
  • EndNote
  • RefWorks
  • TxT
Links
Articles
Latest Articles
Most Read
Collections
Updates
Events
News
Multimedia
About
About Us
Contact
No. 86 Xueyuan South Road, Haidian District, Beijing
100081
010-62199257
qkjq@cast.org.cn

Copyright © 2025 China Association for Science and Technology. All rights reserved. For all open access content, the relevant licensing terms apply.
Sponsored by the Office of the Leading Group for Cybersecurity and Informatization of CAST, and supported by Science and Technology Review Publishing House