Isolation and characterization of antihypertensive peptides from soy bean protein

Proteins and peptides are the most diverse biomolecules found in nature and make our interest due to their wide applications in food and pharmaceutical industry. Angiotensin Converting Enzyme (ACE) plays a major role in controlling blood pressure. The inhibition of ACE with peptides is a main target in the regulation of hypertension. The objective of the present study was to investigate the therapeutic potential of soy bean. This was accomplished by isolation of ACE inhibitory peptides using response surface methodology (RSM) and characterization of these bioactive peptides by mass spectrometry. 31 hydrolyzed fractions were isolated and evaluated for their ACE inhibition potential. Hydrolyzed fraction having highest ACE inhibitory activity was characterized by liquid chromatography-mass spectrometry (LC-MS) technique. RSM results showed maximum ACE inhibition potential (64%) by hydrolyzate was obtained at 45 ºC temperature, pH 8.0, E/S 0.2 in 2 hours hydrolysis time. Results of LC-MS analysis revealed Ser-Gly, Ser-Pro, Met-Ala, His-Ala, Lys-Pro, Phe-Thr, Met-Leu, Pro-Arg, Ala-Pro-Val, Pro-Ala-Leu, Val-Met-Gly, Pro-Leu-Val, Pro-Pro-Gln, His-Arg-Gly, Ser-Phe-Val-Leu, Ala-Val-His-Try, Arg-Thr-Val-Arg, His-His-Tyr-Leu-Val, Asp-Gly-Ala-Cys-Ser-Ala-Asn and MetVal-Thr-Gly-Pro-Gly-Cys-His bioactive peptides in hydrolyzed fraction of soy bean. Our data provide evidence that response surface methodology is a good approach for isolation of antihypertensive bioactive peptides with more potent activity as nutraceuticals or pharmaceuticals. Therefore soy bean can be use for industrial production of pharmaceutical grade natural medicines for handling high blood pressure.

Nutrigenomics as a tool in the prevention of lipotoxicity: the case of soy protein

Abstract Obesity is associated with an increase of several metabolic disorders leading to the development of diseases such as type 2 diabetes and cardiovascular disease. This is due in part to the ectopic accumulation of triglycerides in organs that are non-adipose tissues, leading to lipotoxicity. Particularly, in the liver, the accumulation of lipids, mainly of triglycerides, leads to the formation of fatty liver. The accumulation of lipids in skeletal muscle and pancreas associates with insulin resistance and a decrease in insulin secretion, respectively. In addition, it has been suggested that dysbiosis of the gut microbiota can contribute to the process of lipid accumulation in non-adipose tissues, especially in the liver. The aim of the present review is to highlight the mechanisms associated with the development of lipotoxicity, and how with the advances in nutrigenomics, it is now possible to understand the molecular mechanisms by which some nutrients can attenuate the ectopic accumulation of triglycerides in non-adipose tissues. Particularly, we emphasize research conducted on the molecular mechanisms of action of soy protein and some of its isoflavones, and how these can reduce lipotoxicity by preventing the accumulation of lipids in the liver, skeletal muscle, and pancreas, as well as their role on the gut microbiota to attenuate the development of fatty liver. Thus, nutrigenomics is opening new dietary strategies based on several functional foods that can be used to ameliorate the pathologies associated with lipotoxicity.

Nitric oxide enhancement and blood pressure decrease in patients with metabolic syndrome using soy protein or fish oil / Aumento de óxido nítrico e diminuição da pressão sanguínea em pacientes com síndrome metabólica em uso de proteína de soja ou óleo de peixe

OBJECTIVE: To verify the effects of fish oil and soy on nitric oxide (NO) and blood pressure in patients with metabolic syndrome (MS). SUBJECTS AND METHODS: Sixty women with MS were investigated in a parallel randomized design study. The first group maintained their usual diet; the second group received 25 g/day of soy; the third group received 3 g/day of n-3 fatty acids, and the fourth group the same amount previously cited of n-3 fatty acids and soy. RESULTS: Serum nitric oxide metabolites showed significant increase after 90 days in the fish oil and soy groups. Systolic pressure reduced after 45 days of treatment with fish oil, whereas diastolic pressure decreased significantly throughout the study in the soy group. CONCLUSIONS: NO increase and blood pressure reduction with fish oil or soy protein reinforce the importance of the influence of NO on blood pressure in patients with MS.

OBJETIVO: Verificar o efeito do óleo de peixe e soja sobre o óxido nítrico (NO) e a pressão arterial em pacientes com síndrome metabólica (SM). SUJEITOS E MÉTODOS: Sessenta mulheres com SM foram avaliadas em estudo paralelo randomizado. O primeiro grupo manteve sua dieta habitual, o segundo grupo recebeu 25 g/dia de soja, o terceiro grupo recebeu 3 g/dia de ácidos graxos n-3 e o quarto grupo, a mesma quantidade citada anteriormente de ácidos graxos n-3 e soja. RESULTADOS: Os níveis séricos de metabólitos de NO foram significativamente superiores após 90 dias de intervenção com soja ou óleo de peixe. Somente o grupo que recebeu o óleo de peixe apresentou redução na pressão sistólica após 45 dias. Já a soja reduziu a pressão diastólica em 45 e 90 dias. CONCLUSÕES: O aumento de NO e a redução da pressão arterial com óleo de peixe ou proteína de soja reforçam a influência do óxido nítrico sobre a pressão arterial em pacientes com SM.

Effects of protein quality on appetite and energy metabolism in normal weight subjects / Efeitos da qualidade proteica no apetite e metabolismo energético de indivíduos eutróficos

OBJECTIVE: The purpose of this study was to compare the effects of consumption of different protein sources on food intake and energy expenditure in normal weight subjects. SUBJECTS AND METHODS: Breakfast preparations (casein, soy protein, whey protein or control) were ingested during seven consecutive days. Appetite, food intake, and energy expenditure were assessed. RESULTS: Casein consumption led to a lower energy intake than whey protein. There was lower energy intake on day 7 than on day 1 of the casein session. Soy protein preparations resulted in higher diet induced thermogenesis (DIT) than in control preparations. The respiratory quotient (RQ) obtained in the whey protein session was lower than the control and soy protein sessions. CONCLUSION: These results suggest that the consumption of different protein types leads to distinct effects on satiety (casein), DIT (soy protein), and/or RQ (whey protein).

OBJETIVO: O objetivo deste estudo foi comparar os efeitos do consumo de diferentes fontes proteicas na ingestão alimentar e gasto energético em indivíduos eutróficos. SUJEITOS E MÉTODOS: Preparações (caseína, proteína da soja, proteína do soro de leite ou controle) foram ingeridas no desjejum, durante sete dias consecutivos. RESULTADOS: A caseína resultou em menor ingestão calórica do que o soro de leite. Houve uma menor ingestão calórica no último dia da sessão da caseína em relação ao primeiro dia. Preparações contendo proteína da soja resultaram em maior termogênese induzida pela dieta (TID) em comparação às preparações controle. O cociente respiratório (CR) obtido na sessão do soro de leite foi menor que na sessão controle e da proteína da soja. CONCLUSÃO: Esses resultados sugerem que o consumo de diferentes tipos de proteínas resulta em efeitos distintos na saciedade (caseína), TID (proteína da soja) e/ou CR (proteína do soro).

Effects of isoflavones on the skin of postmenopausal women: a pilot study

OBJECTIVE: The aim of this study was to evaluate the effects of isoflavones on the skin of postmenopausal women. DESIGN: A prospective study was performed with 30 postmenopausal women before and immediately after the end of treatment with 100 mg/day of an isoflavones-rich, concentrated soy extract for six months. A skin punch was performed in the gluteal region for sample collection before and immediately after the treatment program. Morphometric determination of epidermal thickness, the papillary index (wrinkling), and the amount of dermal elastic and collagen fibers was assessed. In addition, the number of blood vessels in the sample was also evaluated. The paired Student¡¯s t-test was used for statistical analysis (P ¡Ü 0.05). RESULTS: Isoflavone treatment resulted in a 9.46% increase in the thickness of the epidermis in 23 patients. In addition, the papillary index was reduced in 21 women. The papillary index was inversely proportional to skin wrinkling, i.e., there were a large number of papillae after treatment. The amount of collagen in the dermis was increased in 25 women (86.2%). In 22 women (75.8%) we observed that the number of elastic fibers increased. The number of dermal blood vessels was significantly increased in 21 women.CONCLUSION: Our data show that the use of a concentrated, isoflavone-rich soy extract during six consecutive months caused significant increases in epithelial thickness, the number of elastic and collagen fibers, as well as the blood vessels.

Hepato and reno protective action of Calendula officinalis L. flower extract.

Flower extract of C. officinalis L. was evaluated for its protective effect against CCl4 induced acute hepatotoxicity and cisplatin induced nephrotoxicity. The activities of serum marker enzymes of liver injury like glutamate pyruvate transaminase (SGPT), glutamate oxaloacetate transaminase (SGOT) and alkaline phosphatase (ALP) which were increased by CCl4 injection was found to be significantly reduced by the pretreatment of the flower extract at 100 and 250 mg/kg body weight. The lipid peroxidation in liver, the marker of membrane damage and the total bilirubin content in serum were also found to be at significantly low level in the extract pretreated group, indicating its protective role. The kidney function markers like urea and creatinine were significantly increased in cisplatin treated animals. However, their levels were found to be lowered in the extract pretreated groups (100 and 250 mg/kg body weight). Moreover, cisplatin induced myelosuppression was ameliorated by the extract pretreatment. Treatment with the extract produced enhancement of antioxidant enzymes--superoxide dismutase and catalase and glutathione. Results suggest a protective role of the flower extract of C. officinalis against CCl4 induced acute hepatotoxicity and cisplatin induced nephrotoxicity. Extract has been found to contain several carotenoids of which lutein, zeaxanthin and lycopene predominates. Possible mechanism of action of the flower extract may be due to its antioxidant activity and reduction of oxygen radicals.

Antiatherogenic and antiperoxidative effects of garlic and soy proteins in alcohol fed rats.

Rats fed with alcohol (18%) at 3.76 g/day for 45 days showed significant reduction in body weight, glutathione (GSH) content and activities of superoxide dismutase (SOD) and catalase in liver. Lecithin cholesterol acyltransferase (LCAT) in plasma, levels of HDL cholesterol in serum, hepatic bile acid production and fecal excretion of neutral sterols also showed significant reduction. Simultaneous feeding of garlic protein (GP) or soy protein (SP) (500 mg/kg body weight/day for 45 days) to alcohol fed groups increased each of above parameters significantly towards normal values. Increase in GSH content and catalase activity in liver, was significantly higher for SP treated group than for GP treated group. However, increase in plasma LCAT was significantly higher for GP treated group than for SP treated group. Alcohol fed rats showed significant increase in liver weight, serum and tissue cholesterol, serum triacylglycerol (TAG), phospholipids (PL) and free fatty acid (FFA) levels and activity of HMGCoA reductase in liver and intestine. Lipid peroxidation, glucose-6 phosphate dehydrogenase (G6PD), glutathione peroxidase (GPx) and glutathione reductase (GR) in liver and incorporation of labeled acetate into liver cholesterol also showed significant increase. GP and SP treated rats showed decrease in these values towards normal. GP feeding showed a better effect than SP in lowering serum and heart total cholesterol, and in maintaining GPx at near normal level, while SP feeding showed a better effect in lowering serum FFA level and maintaining GR activity at near normal level. In suppressing incorporation of labeled acetate into serum cholesterol, GP feeding showed a better effect than SP. Antiatherogenic and antiperoxidative effects of these proteins may be due to lower lysine/arginine ratio.

Effects of isoflavone on oxidative stress parameters and homocysteine in postmenopausal women complaining of insomnia

Sleep disorders have an increased incidence after menopause. The objective of this work was to evaluate the effects of isoflavone on some oxidative stress markers in postmenopausal women complaining of insomnia. Women aged between 50-65 years (n=38) were recruited and assigned to a double-blind placebo controlled study for 4 months. The treated group received 100 mg/day of isoflavones. Blood collections were conducted on three different occasions to assess total glutathione; superoxide dismutase and catalase in erythrocytes; lipid peroxidation; and homocysteine plasma concentrations. No differences between the groups were observed. However, all the patients seem to improve their oxidative stress status and homocysteine concentration after treatment. Superoxide dismutase activity was correlated with age and time of menopause at the beginning of the treatment, but these correlations were no longer observed by the end of the study. Soy isoflavones were not able to overcome the placebo effect for either oxidative stress parameters or homocysteine concentrations.

[ effects of soy bean on serum homocysteine, Lp[a] and lipoproteins in hyperlipidemic postmenopausal women]

Because of undesirable serum lipoprotein profiles, postmenopausal women are at risk of cardiovascular disease. Soy protein may help protect against these risk factors although its effect on homocysteine and Lp [a] is not clear; however, the effects of soy bean in hyperlipidemic postmenopausal women has not been assessed independently yet. The aim of this study was to determine the effects of soy bean on serum concentration of homocysteine, Lp[a] and lipoproteins in hyperlipidemic postmenopausal women. In a randomised clinical trial with parallel design, 34 hyperlipidemic postmenopausal women were randomly assigned to group 1 and given 130 g soy bean containing 50 g/d soy protein and 159/9mg isoflavones, while group 2 continued their usual diet for 10 weeks [controls]. Serum lipoproteins, homocysteine and Lp [a] were measured at baseline and after the 10[th] week. There were significant decreases in Lp [a] LDL-c, TC, LDL-c/HDL-c, TG/HDL-c, TC/HDL-c in group 1, as compared to group 2, after ten weeks of study [p<0.05, p<0.001, p<0.01, p<0.01, p<0.05 and p<0.05 respectively] at the end of the study. Similarly, Lp[a], homocysteine, LDL-c, TC, LDL-c/HDL-c, TG/HDL-c, TC/HDL were significantly decreased [p<0.01, p<0.05 p<0.01, p<0.01, p<0.01, p<0.05, p<0.01] at the end of the study, compared with initial values of the soy group. We can conclude that Soy protein reduces CVD risk in postmenopausal women because of both modest reductions in serum lipoproteins and Lp[a] in hyperlipidemic postmenopausal women

Regulación del metabolismo del colesterol y ácidos grasos en el síndrome nefrótico experimental por las proteínas que se unen a los elementos regulatorios de esteroles (SREBP's): efecto de la soya / Metabolism of cholesterol and fatty acids in nephrotic syndrome and its regulation by sterol regulatory element brinding proteins (SREBP's): Effect of soy protein consumption

El síndrome nefrótico (SN) cursa con hiperlipidemia. Se conoce que la biosíntesis del colesterol y de los ácidos grasos es regulada por los factores transcripcionales que se unen a los elementos de respuesta a esteroles (SREBP's). El consumo de proteína de soya disminuye la concentración de estos lípidos, aunque su mecanismo de acción no es del todo conocido. El objetivo de este estudio fue conocer si el consumo de la proteína de soya reduce los niveles de colesterol y triglicéridos a través de una regulación de las SREBP 's. Se estudiaron ratas Wistar macho con SN experimental por 64 días. Se observó que las concentraciones plasmáticos de colesterol y triglicéridos plasmáticos, así como de la proteinuria eran significativamente menores en las ratas alimentadas con proteína de soya que aquellas que consumían caseína. Estos cambios se asociaron con disminución de la expresión del ARNm SREBP 1 y de las enzimas de la síntesis de ácidos grasos. Los análisis por Western Blot revelaron que en los núcleos de hepatocitos obtenidos de ratas alimentadas con proteína de soya hubo menor presencia del factor transcripcional SREBP 1. Los resultados de este estudio indican que el consumo de proteína de soya produce efectos benéficos durante el síndrome nefrótico.

Hyperlipidemia occurs during nephrotic syndrome (NS). It is known that cholesterol and fatty acid biosynthesis is controlled by the transcription factors sterol regulatory element binding proteins (SREBPs). Soy protein consumption reduces the concentration of these lipids, although its mechanism of action is not well known. The aim of the present study was to establish whether soy protein consumption reduces cholesterol and triglycerides levels by regulating of SREBPs. Male Wistar rats with experimental NS were studied for 64 days. The results showed that rats fed with soy protein had significantly lower plasma cholesterol and triglyceride concentrations as well as proteinuria than rats fed with casein diet. These decrements were associated with a decrease in the expression of SREBP 1 and fatty acid biosynthetic enzymes. In addition, Western blot analysis revealed that in nuclear extracts from hepatocytes of rats fed with soy protein, there was a lower concentration of SREBP 1 than in rats fed with casein. The results of this study indicate that consumption of a soy protein diet has beneficial effects on nephrotic syndrome.

Efeito da fibra do polissacarídeo de soja no peso e na umidade das fezes de ratos em fase de crescimento / The effect of soy polysaccharide fiber on fecal weight and humidity in growing rats

OBJETIVO: Comparar, em ratos, o efeito do polissacarídeo de soja em relação a celulose e fórmula de soja sem fibra alimentar sobre o peso e a umidade fecal. MÉTODOS: Foram utilizados 12 ratos Wistar machos, com 21 dias de vida, distribuídos em três grupos, que receberam ração AIN-93G variando-se a fonte de fibra alimentar: grupo polissacarídeo, com 5 por cento de fibra na forma de polissacarídeo de soja; grupo fórmula de soja, com 5 por cento de fórmula de soja sem fibra; e grupo celulose, com 5 por cento de fibra na forma de celulose. As fezes foram coletadas em três períodos de 72 horas iniciados no 7º, 17º e 27º dia. As mesmas foram pesadas frescas e após secagem em estufa a 105 ºC até peso constante. A umidade foi calculada usando a fórmula [(peso fecal úmido - peso fecal seco)/peso fecal úmido] x 100. RESULTADOS: As somas do peso fecal úmido nos grupos polissacarídeo, fórmula de soja e celulose foram, respectivamente: 17,372±4,743 g; 6,045±0,619 g; 16,012±2,600 g (p = 0,001), observando-se diferença estatisticamente significante dos grupos celulose e polissacarídeo em relação ao grupo fórmula de soja. Não houve diferença estatisticamente significante entre os grupos celulose e polissacarídeo. Para o peso seco, na mesma ordem, observou-se: 6,463±1,177 g; 2,909±0,277 g; 10,068±1,085 g (p < 0,001), com diferença estatisticamente significante entre todos os grupos. CONCLUSAO: Os animais que receberam fórmula de soja apresentaram peso fecal úmido e seco inferior ao dos outros dois grupos, enquanto o polissacarídeo de soja determinou umidade fecal superior à da celulose, provavelmente por maior fermentação no cólon.

Avaliação biológica de soja com baixas atividades de inibidores de tripsina e ausência do inibidor Kunitz / Biological evaluation of soybean line with low trypsin inhibitor activities

The soybean cultivar BR 36 with conventional levels of trypsin inhibitors activity and the soybean line BRM95-5262, which was genetically selected to contain low activity of trypsin inhibitors were used for biological assays with rats. BR 36 and BRM95-5262 contained 40 and 20, and 30 and 20 of relative residual activity of trypsin inhibitors, respectively. The mean values of PER and NPR showed that treatments with crude soybeans were minor than treatments with soybean thermically processed. However, the treatments of thermically processed soybean did not showed significative differences (p > or = 0.05). When the trypsin inhibitors activity were 8.61 and 8.44 UIT/mg of samples or 20 and 30 of relative residual activity of cultivar BR 36 and line BRM95-5262, respectively, it was observed that mean values of PER and NPR were not significatives. The mean values of CDA and CDV of treatments with crude soybeans were minor than treatment with casein and similar to the treatments with soybean thermically processed. So, it can be concluded that the biological evaluation obtained with soybean protein were dependent of initial trypsin inhibitors activities and of its respective thermical treatment. There was advantage in the use of BRM95-5262 soybean line with low trypsin inhibitors activity.