Vasorelaxing and antihypertensive activities of synthesized peptides derived from computer-aided simulation of pepsin hydrolysis of yam dioscorin
© Lin et al.; licensee Springer 2014
Received: 11 April 2014
Accepted: 16 May 2014
Published: 7 June 2014
We reported that yam dioscorin and its peptic hydrolysates exhibited ACE inhibition and antihypertensive effects on SHRs, however, the active peptides are not really isolated until now. Using ACE inhibitory screenings, two penta-peptides, KTCGY and KRIHF, were selected for ex vivo and in vivo experiments.
KTCGY, KRIHF, and captopril were shown to have similar vasodilating effects against phenylephrine (PE)-induced tensions in rat endothelium-dependent thoracic aortic rings, however, KTCGYKTCGY (two-repeated KTCGY) and TCGYTCGY (two-repeated TCGY) were showed endothelium-independent vasodilating effects against PE-induced tensions. KTCGY, KRIHF (10 or 20 mg/kg), and captopril (10 mg/kg) were used to evaluate antihypertensive activity during 24-h after a single oral administration to spontaneously hypertensive rats (SHRs). The KTCGY and KRIHF showed significantly different and reduced the systolic blood pressure of SHRs compared to the blank.
These results suggest that KTCGY and KRIHF may contribute important roles in yam dioscorin for regulating blood pressure in vivo.
Processed yam tuber is a traditional Chinese medicine used as a juvenescent substance, with no apparent side effects resulting from long-term use, which is ranked the top levels in Chinese Pharmacopoeia. Yams (Dioscorea spp., Dioscoreaceae) are an important tuber crop in Africa, Asia, and Middle and South America and are also a staple food in the Caribbean. Yam dioscorin and its peptic hydrolysates have been reported to exhibit several biological activities in vitro and in vivo (Lu et al. ), including antioxidant (Hou et al. ; Liu et al. ; Han et al. , [2014a],[b]), antihypertensive (Hsu et al. ; Lin et al. ; Liu et al. [2009a]), immunomodulatory (Liu et al. ; Liu et al. [2009b]), and enzyme activities (Hou et al. [1999a], [b]; Hou et al. ).
The untreated high blood pressure is considered to be the central factor in stroke which accounts approximately 33% deaths (Mark and Davis ). There are several classes of pharmacological agents which have been used in the treatment of hypertension, and one class of antihypertensive drugs known as angiotensin I converting enzyme (ACE) inhibitors is associated with a low rate of adverse side effects and is the preferred class of antihypertensive agents when treating patients with concurrent secondary diseases or cardiovascular diseases (Zaman et al. ). ACE (peptidyldipeptide hydrolase, EC 18.104.22.168) is a dipeptide-liberating Zn-containing exopeptidase, which removes a dipeptide from the C-terminus of angiotensin I to form angiotensin II, a very hypertensive compound. Several ACE inhibitory peptides were isolated from food proteins and exhibited generally to reduce blood pressures of SHRs (Martínez-Maqueda et al. ). Fujita et al. () found that the octapeptides of FFGRCVSP (IC50 = 0.4 μM) and ERKIKVYL (IC50 = 1.2 μM) were potent ACE inhibitors, but none of them were effective in spontaneously hypertensive rats (SHRs) to reduce the blood pressure. These potential ACE inhibitory peptides were further hydrolyzed by the rat’s gastrointestinal proteases and then lose their antihypertensive effects on SHR in vivo. We reported that yam dioscorin and its peptic hydrolysates exhibited ACE inhibitory activity (Hsu et al. ) and antihypertensive activity (Lin et al. ) using SHRs as models, however, the active peptides are not really isolated until now. In this study, using angiotensin converting enzyme inhibition as preliminary screenings, two out of twenty-three synthesized peptides from a computer-aided simulation of pepsin hydrolysis of yam dioscorin were selected for further ex vivo and in vivo experiments and captopril was used for comparisons. It is suggested that KTCGY and KRIHF show vasodilating effects and can reduce SHR’s systolic blood pressure (SBP) which may contribute important roles in yam dioscorin for regulating blood pressure in vivo.
The 23 peptides for ACE inhibitory activity screenings were synthesized by Mission Biotech. (Taipei, Taiwan). Peptides for ex vivo and in vivo experiments used in this study (KTCGY, KTCGYKTCGY, TCGYTCGY, and KRIHF) were synthesized by Shanghai Hanhong Chemical Co., Ltd (Shanghai, China) or Shanghai Mocell Biotech Co., Ltd (Shanghai, China). The purity of each peptide was determined to be greater than 97% by performing HPLC chromatography and mass spectroscopy. Captopril was purchased from Calbiochem Co. (CA, USA). N-(3-[2-furyl]acryloyl]-Phe-Gly-G ly (FAPGG), ACE (I unit, rabbit lung), dimethyl sulfoxide (DMSO) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Other chemicals and reagents were from Sigma Chemical Co. (St. Louis, MO, USA).
Computer-aided simulation of pepsin hydrolysis of yam dioscorin
The deduced sequences of dioscorin A (UniProtKB/TrEMBL:Q9M519) and dioscorin B (UniProtKB/TrEMBL:Q9M501) were selected from the website of the ExPASy SIB Bioinformatics Resource Portal (http://www.expasy.org/proteomics) to participate in a computer-aided simulation of pepsin hydrolysis (pH > 2) (http://web.expasy.org/peptide_mass/). Twenty-three peptides were selected for syntheses following the general rules which the peptide contained aromatic or branched-chain aliphatic amino acids closed to the COOH-terminus for ACE inhibitions (Cheung et al. ), including (1) KTCGNGME, (2) PPCSE, (3) CDDRVIRTPLT, (4) KTCGY, (5) PPCTE, (6) RDNGVIF, (7) KRIHF, (8) RRDY, (9) RSVF, (10) PTNF, (11) GISW, (12) MGSF, (13) VSIL, (14) HSPA, (15) DPF, (16) RY, (17) RF, (18) NW, (19) RL, (20) GVI, (21) GSL, (22) SY, and (23) GPA.
ACE inhibitory assay screenings
The ACE inhibitory activity was measured following the previous reports with some modifications (Hsu et al. ). Each synthesized peptide was dissolved in DMSO to 4 mM as stocks. The 1 ml, 0.5 mM FAPGG (dissolved in 50 mM Tris–HCl buffer, pH 7.5, containing 0.3 M NaCl) was mixed with 12.2 μl peptide solution, and then twenty μl (20 μU) of commercial ACE (stock solution, 1U/ ml) was added. The 0.1% DMSO solution was used instead of sample solution for blank experiments (ΔAblank/min). The decreased absorbance at 345 nm (ΔAsample/min) was recorded during 90 sec at room temperature. The ACE inhibition (%) was calculated as followed: [1 - (ΔAsample/min ÷ ΔAblank/min)] × 100.
Effects of KTCGY, KTCGYKTCGY, TCGYTCGY, and KRIHF on phenylephrine-induced tensions in rat thoracic aortic ring ex vivo
Effects of four peptides and captopril on vascular tension were examined following the previous method (Wang et al. ). Male Sprague–Dawley rats were killed by decapitation, and sections of the thoracic aorta between the aortic arch and the diaphragm were excised carefully and placed in a Petri dish with oxygenated Krebs’ buffer (120 mM NaCl, 4.5 mM KCl, 2.5 mM CaCl2, 1 mM MgSO4, 27 mM NaHCO3, 1 mM KH2PO4, 10 mM glucose, pH 7.4, with 95% O2, 5% CO2) and gently dissected free of fat and connective tissue. The endothelium was removed by gently rubbing the intimal layer of the vessel with a wire. The isolated aortic rings, 3 to 4 mm in length with or without the endothelium, were fixed isometrically in organ chambers under passive tension (1.8 g) for 60 min. Changes in vascular tension were recorded with a polygraph (Gould, model 2400, Valley View, OH, USA) via a force displacement transducer (Grass FT03, Quincy, MA, USA). After equilibration, the near maximal concentration was induced by PE (0.3 μM). When the rings achieved a stable tension, the acetylcholine (3 μM) was added to the bath to assess endothelial integrity. The relaxing effects of KTCGY, KTCGYKTCGY, TCGYTCGY, KRIHF, and captopril on PE (0.3 μM) pre-contracted endothelium-intact or endothelium-denuded aortic rings were examined. When PE treatments reached a steady plateau (considered as 100%), KTCGY, KTCGYKTCGY, TCGYTCGY, KRIHF, and captopril were then tested at cumulative concentrations (0.1 to 100 μM) for their ability to induce aortic ring relaxations.
Antihypertensive Effects of KTCGY, KRIHF, and captopril in SHRs
Experimental procedures were reviewed and approved by the Institutional Animal Care and Use Committee of Taipei Medical University (LAC-95-0076, LAC-97-0117, and LAC-100-0038). Male SHRs (18–28 weeks old; from National Laboratory Animal Center, Taipei, Taiwan) were housed individually in steel cages kept at 24°C under a 12-h light–dark cycle, with free access to water and a standard mouse/rat chow (Prolab® RMH2500, 5P14 Diet, PMI Nutrition International Brentwood, MO, USA). SHRs were randomly divided into 4 groups (N = 6 of each group), KTCGY or KRIHF at concentration of 10 mg/kg and 20 mg/kg was orally administered to the SHRs, and the SBP were measured after 0, 2, 4, 6, 8, and 24 h by using an indirect tail-cuff blood pressure meter (BP98-A, Softron, Tokyo, Japan). Distilled water (0.5 ml) was administered to the SHRs in the blank group. The captopril (10 mg/kg) was used as the positive control.
Data were expressed as mean ± S.D. For animal experiments, the differences between the blank and the experimental group at the same time was analyzed using Student’s t-test, and the P-value of less than 0.05 (*), 0.01 (**), and 0.001 (***) were recognized as different significantly. The statistical analysis was performed using the SigmaPlot software 10.0.
ACE inhibitory assay screenings
The synthesized peptides derived from computer-aided simulation of pepsin hydrolysis a of yam dioscorin b,c with potential angiotensin converting enzyme inhibitory activities used in this study
Synthesized peptide no.
Molecular mass (Average mass, Da)
dioscorin B (52–56)
dioscorin A (118–122)
dioscorin B (118–122)
Relaxations in rat thoracic aortic ring with or without endothelium ex vivo
Antihypertensive activity in vivo
The present ex vivo and in vivo data showed KTCGY and KRIHF with vasodilating effects against PE-induced endothelium-intact aortic tensions and lowering SHR’s SBP which were derived from a computer-aided simulation of pepsin hydrolysis of the yam tuber dioscorin. The penta-peptide of KTCGY was positioned at amino acid residue 52 to residue 56 of dioscorin B protein (UniProtKB/TrEMBL:Q9M501, Fig. S2), and the penta-peptide of KRIHF was positioned at amino acid residue 118 to residue 122 of dioscorin A protein (UniProtKB/TrEMBL:Q9M519, Fig. S1) and dioscorin B protein (UniProtKB/TrEMBL:Q9M501, Fig. S2). Conlan et al. () first reported two classes of cDNA clones encoding major yam tuber storage proteins from Dioscorea cayenensis and named as dioscorin. We demonstrated that dioscorins from six cultivars of three Dioscorea species accounted over 90% of water-soluble proteins by immune stains (Hou et al. ). The processed yam tuber is a traditional Chinese medicine used as a juvenescent substance, with no apparent side effects after long-term uses; the tuber is involved in the top levels in a famous Chinese pharmacopoeia. It was recently reviewed that yam dioscorin and its peptic hydrolysates exhibited several biological activities in vitro and in vivo (Lu et al. ). It seems that yam or its major protein, dioscorin, may be potentials in developments of several aspects of functional food and needed further investigations.
From the present results of ACE inhibitory activity screenings (Figure 1), the orders of top 5 among 23 synthetic peptides were KTCGY (No.4) > KRIHF (No. 7) > NW (No. 18) > RRDY (No. 8) > RY (No. 16) ≅ SY (No. 22) ≅ CDDRVIRTPLT (No. 3). These potent ACE inhibitory peptides were promised to the general rules which peptides contained aromatic or branched-chain aliphatic amino acids closed to the COOH-terminus for ACE inhibitions (Cheung et al. ). The dioscorin peptic hydrolysates were reported to exhibit ACE inhibitory activity (Hsu et al. ) and antihypertensive activity (Lin et al. ) using SHR as models. Therefore, the synthetic KTCGY and KRIHF were used for ex vivo and in vivo experiments. KTCGY and KRIHF showed endothelium-dependent aortic ring relaxations in PE-induced tensions (Figure 2A, D) which the vasodilating effects were similar to ACE inhibitor, captopril (Figure 2E). The captopril is designed to inhibit ACE, one of important enzymes in renin-angiotensin system, to achieve antihypertension (Rubin et al. ). It was reported that captopril also showed endothelium-dependent rabbit aortic ring relaxations in PE-induced tensions, and the possible action mechanism might be from its superoxide radical scavenging activity of its structural sulfhydryl group (Mittra and Singh ). Hernández-Ledesma et al. () reported that individual amino acids exhibited different oxygen radical absorbance capacity (ORAC) activities, such as Trp [W, 4.649 μmole trolox equivalents (TE)/μmole], Tyr (Y, 1.574 μmole TE/μmole), Met (M, 1.126 μmole TE/μmole), Cys (0.1492 μmole TE/μmole), His (H, 0.073 μmole TE/μmole), and Phe (F, 0.0025 μmole TE/μmole), which a Trp-indole group, Tyr-phenolic group, His-imidazole group, and Cys-sulfhydryl group served as hydrogen donors to contribute ORAC activity. It was reported that the ovokinin (2–7) (a hexa-peptide, RADHPF) isolated from chymotryptic ovalbumin hydrolysates exhibited dose-dependently nitric oxide-stimulated vaso-relaxations in endothelium-intact PE-induced tensions of SHR’s mesenteric artery, however, ovokinin (2–7) showed less effect on ACE inhibition (IC50 higher than 1 mM) (Matoba et al. ). Therefore, we propose that vaso-relaxing peptides of KTCGY and KRIHF in endothelium-intact aortic rings may involve radical scavenging and nitric oxide stimulating activities and need further investigations.
The KTCGYKTCGY and TCGYTCGY showed endothelium-intact and endothelium-denuded vaso-relaxing activities in PE-induced tensions which were different from the action mode of KTCGY of above-mentioned. The ovokinin (an octa-peptide, FRADHPFL) isolated from peptic hydrolysates of egg ovalbumin showed bradykinin B1 receptor antagonist and exhibited endothelium-independent canine mesenteric artery relaxations in prostaglandin F2α-induced tensions (Fujita et al. ). Recently, a series of experiments by Matsui team demonstrated that WH di-peptide showed endothelium-independent SD rat aortic ring relaxations in KCl-induced contractions by suppressing Ca2+ influx (Tanaka et al. ), which the inhibition of extracellular calcium entry into vascular smooth muscle cells by blocking dihydropyridine-like L-type Ca2+ channels (Wang et al. ), and the inhibition against voltage-dependent L-type Ca2+ channels phosphorylation (Kobayashi et al. ). The HRW tri-peptide also showed endothelium-independent vaso-relaxing activities in PE-induced tensions by inhibiting extracellular Ca2+ influx (Tanaka et al. ). Therefore, it is proposed that KTCGYKTCGY and TCGYTCGY with endothelium-independent vaso-relaxing activities in PE-induced tensions might involve Ca2+ influx and Ca2+ channels and needed investigations further.
From the present results of in vivo SHR oral administration, both penta-peptides of KTCGY and KRIHF at doses of 10 and 20 mg/kg exhibited antihypertensive activities by lowering SBP, but not diastolic blood pressure (data not shown), among which KTCGY of 20 mg/kg exhibited the similar lowering SBP profile to captopril of 10 mg/kg after a single oral administration. However, the vaso-relaxing peptide with antihypertensive activity is not necessary for potent ACE inhibition. Therefore, it might loss some antihypertensive peptides from ACE inhibitory screenings in the present study. The RF di-peptide (Kagebayashi et al. ) and IHRF tetra-peptide (Kontani et al. ) isolated from rice glutelin with lower ACE inhibition was reported to exhibit cholecystokinin-dependent vaso-relaxing and antihypertensive activities in SHR, which the RF di-peptide was the same as No. 17 synthesized peptide in Figure 1 of less ACE inhibitory activity in the present study from computer-aided simulation of pepsin hydrolysis of yam dioscorin A (residues of 134–135 and 158–159) and yam dioscorin B (residues of 158–159). However, these results provide evidences to support yam dioscorin after ingestion for blood pressure regulations.
The KTCGY and KRIHF show vasodilating effects and can reduce SHR’s systolic blood pressure which may contribute important roles in yam dioscorin for regulating blood pressure in vivo and will be beneficial for anti-hypertension in functional food preparations.
The authors would like to express thanks to Ministry of Science and Technology, Republic of China (NSC 102-2313-B-038 -004 -MY3) for financial supports.
- Cheung HS, Wang FL, Ondetti MA, Sabo EF, Cushman DW: Binding of peptide substrates and inhibitors of angiotensin-converting enzyme. Importance of the COOH-terminal dipeptide sequence. J Biol Chem 1980, 255: 401–407.PubMedGoogle Scholar
- Conlan RS, Griffiths LA, Napier JA, Shewry PR, Mantell S, Ainsworth C: Isolation and characterisation of cDNA clones representing the genes encoding the major tuber storage protein (dioscorin) of yam ( Dioscorea cayenensis Lam.). Plant Mol Biol 1995, 28: 369–380. 10.1007/BF00020387View ArticlePubMedGoogle Scholar
- Fujita H, Usui H, Kurahashi K, Yoshikawa M: Isolation and characterization of ovokinin, a bradykinin B 1 agonist peptide derived from ovalbumin. Peptides 1995, 16: 785–790. 10.1016/0196-9781(95)00054-NView ArticlePubMedGoogle Scholar
- Fujita H, Yokoyama K, Yoshikawa M: Classification and antihypertensive activity of angiotensin I-converting enzyme inhibitory peptides derived from food proteins. J Food Sci 2000, 65: 564–569. 10.1111/j.1365-2621.2000.tb16049.xView ArticleGoogle Scholar
- Han CH, Liu JC, Fang SU, Hou WC: Antioxidant activities of the synthesized thiol-contained peptides derived from computer-aided pepsin hydrolysis of yam tuber storage protein, dioscorin. Food Chem 2013, 138: 923–930. 10.1016/j.foodchem.2012.11.101View ArticlePubMedGoogle Scholar
- Han CH, Lin YS, Lin SY, Hou WC: Antioxidant and antiglycation activities of the synthesised dipeptide, Asn-Trp, derived from computer-aided simulation of yam dioscorin hydrolysis and its analogue, Gln-Trp. Food Chem 2014, 147: 195–202. 10.1016/j.foodchem.2013.09.109View ArticlePubMedGoogle Scholar
- Han CH, Lin YF, Lin YS, Lee TL, Huang WJ, Lin SY, Hou WC: Effects of yam tuber protein, dioscorin, on attenuating oxidative status and learning dysfunction in D-galactose-induced BALB/c mice. Food Chem Toxicol 2014, 65: 356–363. 10.1016/j.fct.2014.01.012View ArticlePubMedGoogle Scholar
- Hernández-Ledesma B, Dávalos A, Bartolomé B, Amigo L: Preparation of antioxidant enzymatic hydrolysates from alpha-lactalbumin and beta-lactoglobulin. Identification of active peptides by HPLC-MS/MS. J Agric Food Chem 2005, 53: 588–593. 10.1021/jf048626mView ArticlePubMedGoogle Scholar
- Hou WC, Liu JS, Chen HJ, Chen TE, Chang CF, Lin YH: Dioscorin, the major tuber storage protein of yam ( Dioscorea batatas Decne) with carbonic anhydrase and trypsin inhibitor activities. J Agric Food Chem 1999, 47: 2168–2172. 10.1021/jf980738oView ArticlePubMedGoogle Scholar
- Hou WC, Chen HJ, Lin YH: Dioscorins, the major tuber storage proteins of yam ( Dioscorea batatas Decne), with dehydroascorbate reductase and monodehydroascorbate reductase activities. Plant Sci 1999, 149: 151–156. 10.1016/S0168-9452(99)00152-1View ArticleGoogle Scholar
- Hou WC, Chen HJ, Lin YH: Dioscorins from different Dioscorea species all exhibit both carbonic anhydrase and trypsin inhibitor activities. Bot Bull Acad Sin 2000, 41: 191–196.Google Scholar
- Hou WC, Lee MH, Chen HJ, Liang WL, Han CH, Liu YW, Lin YH: Antioxidant activities of dioscorin, the storage protein of yam ( Dioscorea batatas Decne) tuber. J Agric Food Chem 2001, 49: 4956–4960. 10.1021/jf010606mView ArticlePubMedGoogle Scholar
- Hsu FL, Lin YH, Lee MH, Lin CL, Hou WC: Both dioscorin, the tuber storage protein of yam ( Dioscorea alata cv. Tainong No. 1), and its peptic hydrolysates exhibited angiotensin converting enzyme inhibitory activities. J Agric Food Chem 2002, 50: 6109–6113. 10.1021/jf0203287View ArticlePubMedGoogle Scholar
- Kagebayashi T, Kontani N, Yamada Y, Mizushige T, Arai T, Kino K, Ohinata K: Novel CCK-dependent vasorelaxing dipeptide, Arg-Phe, decreases blood pressure and food intake in rodents. Mol Nutr Food Res 2012, 56: 1456–1463. 10.1002/mnfr.201200168View ArticlePubMedGoogle Scholar
- Kobayashi Y, Fukuda T, Tanaka M, Matsui T: The anti-atherosclerotic di-peptide, Trp-His, inhibits the phosphorylation of voltage-dependent L-type Ca 2+ channels in rat vascular smooth muscle cells. FEBS Open Bio 2012, 2: 83–88. 10.1016/j.fob.2012.04.005View ArticlePubMedPubMed CentralGoogle Scholar
- Kontani N, Omae R, Kagebayashi T, Kaneko K, Yamada Y, Mizushige T, Kanamoto R, Ohinata K: Characterization of Ile-His-Arg-Phe, a novel rice-derived vasorelaxing peptide with hypotensive and anorexigenic activities. Mol Nutr Food Res 2014, 58: 359–364. 10.1002/mnfr.201300334View ArticlePubMedGoogle Scholar
- Lin CL, Lin SY, Lin YH, Hou WC: Effects of tuber storage protein of yam ( Dioscorea alata cv. Tainong No. 1) and its peptic hydrolyzates on spontaneously hypertensive rats. J Sci Food Agric 2006, 86: 1489–1494. 10.1002/jsfa.2530View ArticleGoogle Scholar
- Liu YH, Liang HJ, Cheng HC, Liu YW, Hou WC: Comparisons of in vitro antioxidant activities of storage proteins in tuber of two Dioscorea species. Bot Stud 2006, 47: 231–237.Google Scholar
- Liu YW, Shang HF, Wang CK, Hsu FL, Hou WC: Immunomodulatory activity of dioscorin, the storage protein of yam ( Dioscorea alata cv. Tainong No. 1) tuber. Food Chem Toxicol 2007, 45: 2312–2318. 10.1016/j.fct.2007.06.009View ArticlePubMedGoogle Scholar
- Liu DZ, Liang HJ, Han CH, Lin SY, Chen CT, Fan M, Hou WC: Feeding trial of instant food containing lyophilised yam powder in hypertensive subjects. J Sci Food Agric 2009, 89: 138–143. 10.1002/jsfa.3420View ArticleGoogle Scholar
- Liu YW, Liu JC, Huang CY, Wang CK, Shang HF, Hou WC: Effects of oral administration of yam tuber storage protein, dioscorin, to BALB/c mice for 21-days on immune responses. J Agric Food Chem 2009, 57: 9274–9279. 10.1021/jf902245kView ArticlePubMedGoogle Scholar
- Lu YL, Chia CY, Liu YW, Hou WC: Biological activities and applications of dioscorins, the major tuber storage proteins of yam. J Tradit Complement Med 2012, 2: 41–46.PubMedPubMed CentralGoogle Scholar
- Mark KS, Davis TP: Stroke: development, prevention and treatment with peptidase inhibitors. Peptides 2000, 21: 1965–1973. 10.1016/S0196-9781(00)00346-6View ArticlePubMedGoogle Scholar
- Martínez-Maqueda D, Miralles B, Recio I, Hernández-Ledesma B: Antihypertensive peptides from food proteins: a review. Food Funct 2012, 3: 350–361. 10.1039/c2fo10192kView ArticlePubMedGoogle Scholar
- Matoba N, Usui H, Fujita H, Yoshikawa M: A novel anti-hypertensinve peptide derived from ovalbumin induces nitric oxide-mediated vasorelaxation in an isolated SHR mesenteric artery. FEBS Lett 1999, 452: 181–184. 10.1016/S0014-5793(99)00587-6View ArticlePubMedGoogle Scholar
- Mittra S, Singh M: Possible mechanism of captopril induced endothelium-dependent relaxation in isolated rabbit aorta. Mol Cell Biochem 1998, 183: 63–67. 10.1023/A:1006854313163View ArticlePubMedGoogle Scholar
- Rubin B, Antonaccio MJ, Horovitz ZP: Captopril (SQ 14 225) (D-3-mercapto-2-methylpropanoyl-L-proline): a novel orally active inhibitor of angiotensin-converting enzyme. Prog Cardiovasc Dis 1978, 21: 183–194. 10.1016/0033-0620(78)90024-5View ArticlePubMedGoogle Scholar
- Tanaka M, Tokuyasu M, Matsui T, Matsumoto K: Endothelium-independent vasodilation effect of di- and tri-peptides in thoracic aorta of SpragueDawley rats. Life Sci 2008, 82: 869–875. 10.1016/j.lfs.2008.02.001View ArticlePubMedGoogle Scholar
- Tanaka M, Watanabe S, Wang Z, Matsumoto K, Matsui T: His-Arg-Trp potently attenuates contracted tension of thoracic aorta of Sprague–Dawley rats through the suppression of extracellular Ca 2+ influx. Peptides 2009, 30: 1502–1507. 10.1016/j.peptides.2009.05.012View ArticlePubMedGoogle Scholar
- Wang GJ, Shum AY, Lin YL, Liao JF, Wu XC, Ren J, Chen CF: Calcium channel blockade in vascular smooth muscle cells: major hypotensive mechanism of S-petasin, a hypotensive sesquiterpene from Petasites formosanus . J Pharmacol Exp Ther 2001, 297: 240–246.PubMedGoogle Scholar
- Wang Z, Watanabe S, Kobayashi Y, Tanaka M, Matsui T: Trp-His, a vasorelaxant di-peptide, can inhibit extracellular Ca 2+ entry to rat vascular smooth muscle cells through blockade of dihydropyridine-like L-type Ca 2+ channels. Peptides 2010, 31: 2060–2066. 10.1016/j.peptides.2010.07.013View ArticlePubMedGoogle Scholar
- Zaman MA, Oparil S, Calhoun DA: Drugs targeting the renin-angiotensin-aldosterone system. Nat Rev Drug Discov 2002, 1: 621–636. 10.1038/nrd873View ArticlePubMedGoogle Scholar
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