Recent Advances in In Vitro and In Vivo Studies of Antioxidant, ACE-Inhibitory and Anti-Inflammatory Peptides from Legume Protein Hydrolysates.

Consumption of legumes has been shown to enhance health and lower the risk of cardiovascular disease and specific types of cancer. ACE inhibitors, antioxidants, and synthetic anti-inflammatories are widely used today; however, they have several undesirable side effects. Thus, researchers have focused on finding ACE inhibitors, antioxidant, and anti-inflammatory peptides from natural sources, such as legumes. Recently, in vitro and in vivo research has shown the bioactive peptides generated from legume protein hydrolysates, such as antioxidant, anti-hypertensive, anticancer, anti-proliferative, anti-inflammatory, etc., in the context of different disease mitigation. Therefore, this review aims to describe the recent advances in in vitro and in vivo studies of antioxidant, anti-hypertensive and anti-inflammatory peptides isolated from legume-derived protein hydrolysates. The results indicated that antioxidant legumes peptides are characterized by short-chain sequence amino acids and possess anti-hypertensive properties by reducing systolic blood pressure (SBP) in spontaneously hypertensive rats (SHR).

Inhibition of Endoplasmic Reticulum Stress Improves Acetylcholine-Mediated Relaxation in the Aorta of Type-2 Diabetic Rats.

Endoplasmic reticulum (ER) stress contributes to insulin resistance and macro- and microvascular complications associated with diabetes. This study aimed to evaluate the effect of ER stress inhibition on endothelial function in the aorta of type-2 diabetic rats. Type-2 diabetes was developed in male Sprague-Dawley rats using a high-fat diet and low-dose streptozotocin. Rat aortic tissues were harvested to study endothelial-dependent relaxation. The mechanisms for acetylcholine-mediated relaxation were investigated using pharmacological blockers, Western blotting, oxidative stress, and inflammatory markers. Acetylcholine-mediated relaxation was diminished in the aorta of diabetic rats compared to control rats; supplementation with TUDCA improved relaxation. In the aortas of control and diabetic rats receiving TUDCA, the relaxation was mediated via eNOS/PI3K/Akt, NAD(P)H, and the KATP channel. In diabetic rats, acetylcholine-mediated relaxation involved eNOS/PI3K/Akt and NAD(P)H, but not the KATP channel. The expression of ER stress markers was upregulated in the aorta of diabetic rats and reduced with TUDCA supplementation. The expression of eNOS and Akt were lower in diabetic rats but were upregulated after supplementation with TUDCA. The levels of MDA, IL-6, and SOD activity were higher in the aorta of the diabetic rats compared to control rats. This study demonstrated that endothelial function was impaired in diabetes, however, supplementation with TUDCA improved the function via eNOS/Akt/PI3K, NAD(P)H, and the KATP channel. The improvement of endothelial function was associated with increased expressions of eNOS and Akt. Thus, ER stress plays a crucial role in the impairment of endothelial-dependent relaxation. Mitigating ER stress could be a potential strategy for improving endothelial dysfunction in type-2 diabetes.

Lipid peroxidation in the descending thoracic aorta of rats deprived of REM sleep using the inverted flowerpot technique.

NEW FINDINGS: What is the central question of this study? Deprivation of rapid eye movement (REM) sleep is associated with increased oxidative stress, but its effects on the blood vessels are poorly documented. We investigated whether REM sleep deprivation induces oxidative stress and causes lipid peroxidation in the aorta. What is the main finding and its important? We demonstrate that REM sleep deprivation induces oxidative stress and mediates lipid peroxidation in the aorta. This can cause endothelial changes and increased blood pressure. These findings will contribute to the growing body of literature on the mechanism underlying the effects of sleep deprivation on cardiovascular disease. ABSTRACT: Oxidative stress-mediated lipid peroxidation is a known cause of endothelial injury or dysfunction. Deprivation of rapid eye movement (REM) sleep is associated with oxidative stress. To date, the pathogenesis of increased blood pressure after sleep deprivation remains poorly understood, particularly in the REM sleep phase. Our aim was to investigate the effects of REM sleep deprivation on blood vessels in the REM sleep-deprived rat model. Twenty-eight male Sprague-Dawley rats were divided into four equal groups: free-moving control rats, rats deprived of REM sleep for 72 h (REMsd), tank control rats and 72 h sleep-recovered rats after 72 h of REM sleep deprivation. The rats were deprived of REM sleep using the inverted flowerpot technique. Food consumption, body weight gain and systolic blood pressure were monitored. At the end of the experiment, the descending thoracic aorta was isolated for the measurement of oxidative stress markers. Despite a significant increase in food consumption in the REMsd group compared with the other groups, there was a significant reduction in body weight gain. Systolic blood pressure also showed a significant increase in the REMsd group compared with the other groups. Superoxide dismutase activity was significantly lower and malondialdehyde concentrations significantly higher in the REMsd group compared with the other groups. Increased levels of malondialdehyde are suggestive of lipid peroxidation in the blood vessels, and oxidative stress may be attributed to the initiation of the process. The changes after REM sleep deprivation revert during sleep recovery. In conclusion, the findings of the present study provide convincing evidence that REM sleep deprivation induced lipid peroxidation, leading to endothelial damage.

Knowledge and Attitude about Stem Cells and Their Application in Medicine among Nursing Students in Universiti Sains Malaysia, Malaysia.

BACKGROUND: Stem cell research has been extensively explored worldwide to enhance human health in medical setting. Nevertheless, there is currently no full understanding of the stem cell knowledge and attitude levels among student nurses in Malaysia. This study aimed to assess the level of stem cell knowledge, attitude toward stem cell application in medicine, and its association with years of education, among Universiti Sains Malaysia (USM) undergraduate nursing students. METHODS: A cross-sectional study (n = 88) was conducted using self-administered questionnaire consisted of demographic information, stem cells knowledge and attitude statements. Data was analysed using Statistical Package Social Software 20.0. RESULTS: The majority of participants (92%) had moderate knowledge score about stem cells. Many students (33%) worried that stem cell application might cause a harm to humanity yet had a positive (76.1%) attitude towards its therapeutic potential (45.5%). Poor correlation between knowledge and attitude (r = 0.08) indicated that acceptance towards stem cell is not solely based on the knowledge level but also on other factors including religion and culture. CONCLUSION: Therefore, this study suggests that various educational programs on stem cell should be implemented considering the religion, cultural, social, and behavioural determinants in the population to improve stem cell knowledge and encourage a more positive attitude towards stem cells in medicine among these nursing students.

Hepatic and renal effects of oral stingless bee honey in a streptozotocin-induced diabetic rat model.

BACKGROUND: Diabetes is known damage the liver and kidney, leading to hepatic dysfunction and kidney failure. Honey is believed to help in lowering the blood glucose levels of diabetic patients and reducing diabetic complications. However, the effect of stingless bee honey (SBH) administration in relieving liver and kidney damage in diabetes has not been well-studied. AIM: To investigate the effect of SBH administration on the kidney and liver of streptozotocin-induced (STZ; 55 mg/kg) diabetic Sprague Dawley rats. METHODS: The rats were grouped as follows (n = 6 per group): non-diabetic (ND), untreated diabetic (UNT), metformin-treated (MET), and SBH+metformin-treated (SBME) groups. After successful diabetic induction, ND and UNT rats were given normal saline, whereas the treatment groups received SBH (2.0 g/kg and/or metformin (250 mg/kg) for 12 d. Serum biochemical parameters and histological changes using hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) staining were evaluated. RESULTS: On H&E and PAS staining, the ND group showed normal architecture and cellularity of Bowman's capsule and tubules, whereas the UNT and MET groups had an increased glomerular cellularity and thickened basement membrane. The SBH-treated group showed a decrease in hydropic changes and mild cellularity of the glomerulus vs the ND group based on H&E staining, but the two were similar on PAS staining. Likewise, the SBME-treated group had an increase in cellularity of the glomerulus on H&E staining, but it was comparable to the SBH and ND groups on PAS staining. UNT diabetic rats had tubular hydropic tubules, which were smaller than other groups. Reduced fatty vacuole formation and dilated blood sinusoids in liver tissue were seen in the SBH group. Conversely, the UNT group had high glucose levels, which subsequently increased MDA levels, ultimately leading to liver damage. SBH treatment reduced this damage, as evidenced by having the lowest fasting glucose, serum alanine transaminase, aspartate transaminase, and alkaline phosphatase levels compared to other groups, although the levels of liver enzymes were not statistically significant. CONCLUSION: The cellularity of the Bowman's capsule, as well as histological alteration of kidney tubules, glomerular membranes, and liver tissues in diabetic rats after oral SBH resembled those of ND rats. Therefore, SBH exhibited a protective hepatorenal effect in a diabetic rat model.

Lignosus rhinocerotis extract ameliorates airway inflammation and remodelling via attenuation of TGF-ß1 and Activin A in a prolonged induced allergic asthma model.

Allergic asthma is associated with chronic airway inflammation and progressive airway remodelling. The sclerotium of Lignosus rhinocerotis (Cooke) Ryvarden (Tiger Milk mushroom) is used traditionally to treat various illnesses, including asthma in Southeast Asia. This study was carried out to evaluate the effect of L. rhinocerotis extract (LRE) on airway inflammation and remodelling in a chronic model of asthma. The present study investigated the therapeutic effects of LRE on airway inflammation and remodelling in prolonged allergen challenged model in allergic asthma. Female Balb/C mice were sensitised using ovalbumin (OVA) on day 0 and 7, followed by OVA-challenged (3 times/week) for 2, 6 and 10 weeks. LRE (125, 250, 500 mg/kg) were administered by oral gavage one hour after every challenge. One group of mice were left untreated after the final challenge for two weeks. LRE suppressed inflammatory cells and Th2 cytokines (IL-4, IL-5 and IL-13) in BALF and reduced IgE level in the serum. LRE also attenuated eosinophils infiltration and goblet cell hyperplasia in the lung tissues; as well as ameliorated airway remodelling by reducing smooth muscle thickness and reducing the expressions of TGF-ß1 and Activin A positive cell in the lung tissues. LRE attenuated airway inflammation and remodelling in the prolonged allergen challenge of allergic asthma model. These findings suggest the therapeutic potential of LRE as an alternative for the management of allergic asthma.

Etlingera elatior Flower Aqueous Extract Protects against Oxidative Stress-Induced Nephropathy in a Rat Model of Type 2 Diabetes.

Diabetes mellitus (DM) is a known systemic disease with increasing global prevalence and multi-organ complications including diabetic nephropathy (DN). The trend of using medicinal plants in the management of DM is increasing exponentially. Etlingera elatior is a medicinal plant that contains chemicals and antioxidants that delay the oxidation process. However, available data focusing on its use on DN are inconsistent and scarce. This study aims to investigate the antidiabetic and nephroprotective effects of E. elatior flower aqueous extract (EEAE) in a type 2 DM rat (T2DR) model. The T2DR model was developed using a combination of a high-fat diet (HFD) and a low dose of streptozotocin (STZ) at 35 mg/kg. Thirty-two Sprague Dawley male rats were randomly divided into four groups (n = 8): (1) control (normal rat), (2) T2DR (untreated-type 2 diabetic rat), (3) Met (250 mg/kg metformin-treated T2DR), and (4) EEAE (1000 mg/kg EEAE-treated T2DR). All treatments were administered orally for 6 weeks. EEAE significantly reduced fasting blood glucose (FBG), microalbuminuria, serum creatinine, and serum blood urea nitrogen. EEAE also reduced malondialdehyde (MDA) and enhanced the levels of antioxidant markers-superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and total antioxidant capacity (T-AOC). The inflammatory marker (interleukin (IL)-6) and fibrosis markers (transforming growth factor-beta (TGF-ß), and connective tissue growth factor (CTGF)) were significantly decreased in the EEAE-treated group. The T2DR group developed DN, which was characterized by segmental sclerosis of the glomeruli associated with focal tubular atrophy and interstitial fibrosis. Interestingly, the histology of kidney tissue in the EEAE group was preserved. This effect was similar to that of the control drug metformin. In summary, the antidiabetic and nephroprotective effects might be related to the antioxidant properties and anti-inflammatory effects of the EEAE. The antidiabetic activity could be due to the presence of the active compound cyanidin-3-O-glycosides, which is an anthocyanin antioxidant, that is present in the EEAE. E. elatior has the potential to be developed as a natural source of antioxidants that can be used for the prevention or even the treatment of DM. These findings could lead to future research into the therapeutic use of E. elatior in alleviating the progression of DM and thus preventing nephropathy.

Systematic Review of Medicinal Plants Used for Treatment of Diabetes in Human Clinical Trials: An ASEAN Perspective.

Traditionally, there are some medicinal plants believed to treat diabetes, as they have been proven in research studies to possess antidiabetic properties, such as improved insulin sensitivity and hypoglycemic activities, due to their high level of phenolic compounds, flavonoids, terpenoids, alkaloids, and glycosides. We conducted a systematic review to identify potential medicinal plants used during human clinical trials in the Association of Southeast Asian Nation (ASEAN) countries on prediabetic or type 2 diabetic individuals and to potentially identify any bioactive compounds involved in effectively treating symptoms of diabetes such as lowering of blood glucose. A total of 1209 reference titles were retrieved from four selected databases (Science Direct, Scopus, Springer Link, and PubMed) and only three met the inclusion criteria. Upon evaluation of the selected articles, four medicinal plants were identified: turmeric (Curcuma longa), garlic (Allium sativum L.), bitter melon (Momordica charantia), and Rosella flower (Hibiscus sabdariffa L.). Of these, only the bitter melon study did not show any significant change in the blood glucose of participants after intervention. This review demonstrates the limitations in published articles of human clinical trials for medicinal plants' intervention for diabetes. Upon further investigations on the four identified medicinal plants included in the animal studies, the findings showed positive effects in the management of diabetes, such as hyperglycemia. Hence, further testing and standardization of the methods in the studies can be suggested for human clinical trials for reliable data collections such as methods of extract preparation, duration of intervention, and conditions set for the study design.

Current Updates on Potential Role of Flavonoids in Hypoxia/Reoxygenation Cardiac Injury Model.

Clinically, timely reperfusion strategies to re-establish oxygenated blood flow in ischemic heart diseases seem to salvage viable myocardium effectively. Despite the remarkable improvement in cardiac function, reperfusion therapy could paradoxically trigger hypoxic cellular injury and dysfunction. Experimental laboratory models have been developed over the years to explain better the pathophysiology of cardiac ischemia-reperfusion injury, including the in vitro hypoxia-reoxygenation cardiac injury model. Furthermore, the use of nutritional myocardial conditioning techniques have been successful. The cardioprotective potential of flavonoids have been greatly linked to its anti-oxidant, anti-apoptotic and anti-inflammatory properties. While several studies have reviewed the cardioprotective properties of flavonoids, there is a scarce evidence of their function in the hypoxia-reoxygenation injury cell culture model. Hence, the aim of this review was to lay out and summarize our current understanding of flavonoids' function in mitigating hypoxia-reoxygenation cardiac injury based on evidence from the last five years. We also discussed the possible mechanisms of flavonoids in modulating the cardioprotective effects as such information would provide invaluable insight on future therapeutic application of flavonoids.

Lignosus rhinocerotis Cooke Ryvarden ameliorates airway inflammation, mucus hypersecretion and airway hyperresponsiveness in a murine model of asthma.

Lignosus rhinocerotis Cooke. (L. rhinocerotis) is a medicinal mushroom traditionally used in the treatment of asthma and several other diseases by the indigenous communities in Malaysia. In this study, the effects of L. rhinocerotis on allergic airway inflammation and hyperresponsiveness were investigated. L. rhinocerotis extract (LRE) was prepared by hot water extraction using soxhlet. Airway hyperresponsiveness (AHR) study was performed in house dust mite (HDM)-induced asthma in Balb/c mice while airway inflammation study was performed in ovalbumin (OVA)-induced asthma in Sprague-Dawley rats. Treatment with different doses of LRE (125, 250 and 500 mg/kg) significantly inhibited AHR in HDM-induced mice. Treatment with LRE also significantly decreased the elevated IgE in serum, Th2 cytokines in bronchoalveolar lavage fluid and ameliorated OVA-induced histological changes in rats by attenuating leukocyte infiltration, mucus hypersecretion and goblet cell hyperplasia in the lungs. LRE also significantly reduced the number of eosinophils and neutrophils in BALF. Interestingly, a significant reduction of the FOXP3+ regulatory T lymphocytes was observed following OVA induction, but the cells were significantly elevated with LRE treatment. Subsequent analyses on gene expression revealed regulation of several important genes i.e. IL17A, ADAM33, CCL5, IL4, CCR3, CCR8, PMCH, CCL22, IFNG, CCL17, CCR4, PRG2, FCER1A, CLCA1, CHIA and Cma1 which were up-regulated following OVA induction but down-regulated following treatment with LRE. In conclusion, LRE alleviates allergy airway inflammation and hyperresponsiveness, thus suggesting its therapeutic potential as a new armamentarium against allergic asthma.

Is Heart Rate a Confounding Factor for Photoplethysmography Markers? A Systematic Review.

Finger photoplethysmography (PPG) waveform is blood volume change of finger microcirculation that reflects vascular function. Reflection index (RI), stiffness index (SI) and second derivative of photoplethysmogram (SDPPG) are derived from PPG waveforms proposed as cardiovascular disease (CVD) markers. Heart rate (HR) is a known factor that affects vascular function. Individual resting HR variation may affect RI, SI and SDPPG. This review aims to identify studies about the relationship between HR with RI, SI and SDPPG among humans. A literature search was conducted in Medline via the Ebscohost and Scopus databases to find relevant articles published within 11 years. The main inclusion criteria were articles in the English language that discuss the relationship between HR with RI, SI and SDPPG using PPG among humans. The search found 1960 relevant articles but only six articles that met the inclusion criteria. SI and RI showed an association with HR. SDPPG (SDPPG-b/SDPPG-a ratio, SDPPG-d/SDPPG-a ratio, aging index (AGI) and revised aging index (RAGI)) also had an association with HR. Only RI had a considerable association with HR, the association between SI and HR was non-considerable and the association between HR and SDPPG was inconclusive. Further interventional studies should be conducted to investigate this issue, as a variation in resting HR may challenge the validity of PPG-based CVD markers.

Supplementation of Stingless Bee Honey from Heterotrigona itama Improves Antiobesity Parameters in High-Fat Diet Induced Obese Rat Model.

Heterotrigona itama is a common stingless bee species found in Southeast Asia. Studies on the health benefits of its honey are limited in comparison with other stingless bee species. This study examines the antiobesity benefits found in stingless bee honey (SBH) from H. itama. The parameters used to measure the benefits were weight change, morphological structures, and biochemical characteristics. The research was conducted by using rats that were given a high-fat diet (HFD). In total 48 male Sprague Dawley (SD) rats were given a formulated HFD to increase the levels of obesity, the HFD was administered with a value of 0.68 g/cm2. The duration of the treatment was six weeks, and the results show that the induction obesity using the HFD was successful. Following this, the rats were then treated with SBH (at dosages of 1000 mg/kg, 750 mg/kg or 500 mg/kg), with orlistat or with a placebo. Compared with typical obesity treatment methods, the one that used the three dosages of SBH showed a higher reduction in body mass index (BMI), percentage of body weight gain, adiposity index, and relative organ weight (ROW). The levels of liver enzymes (ALT, AST, and ALP) were also significantly lower in SBH-treated groups. The levels of triglycerides and LDL-cholesterol were significantly lower, while the level of HDL-cholesterol was significantly higher in comparison with the control obese group. In terms of morphological structures, the number of adipocyte cells was reduced, and the hepatocytes found in the liver were less prone to rupturing when treated with SBH. In conclusion, the administration of SBH led to an improvement in indicators associated with obesity reduction. SBH also possesses a hepatoprotective potential which can reduce the health risks related to obesity.

Autonomic Receptors and Nitric-Oxide Involvements in Mediating Vasorelaxation Effect Induced by Syzygium polyanthum Leaves Extract.

CONTEXT: Syzygium polyanthum (Wight) Walp leaves are traditionally used by Malays for treating hypertension. Our previous study showed that aqueous extract of S. polyanthum (AESP) and methanolic extract of S. polyanthum (MESP) extracts of S. polyanthum leaves significantly reduced blood pressure of normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). AIMS: This study aimed to investigate their vasorelaxation potential and the possible involvement of autonomic receptors and nitric oxide in mediating their effect. SETTINGS AND DESIGN: Both extracts will be tested on isolated thoracic aorta rings of WKY and SHR. The involvement of autonomic receptors and nitric oxide will be elucidated using respective blockers. MATERIALS AND METHODS: Isolated thoracic aorta rings from WKY and SHR were mounted onto myograph chambers to measure changes in the aorta tension. Increasing concentrations of AESP and MESP, from 1 µg/ml to 10 mg/ml were added onto the myograph chambers. Blockers such as atropine (1 µM), phentolamine (1 µM), propranolol (1 µM), and Nω-nitro-l-arginine methyl ester (100 µM) were preincubated before addition of extracts to check for involvement of muscarinic, α- and ß-adrenergic receptors (AR) as well as nitric oxide, respectively. STATISTICAL ANALYSIS USED: Two-way ANOVA, followed by post hoc Bonferroni test was used, where P < 0.05 (two-tailed) was considered statistically significant. RESULTS: AESP and MESP caused significant vasorelaxations through nitric oxide pathway. The former was mediated through α-AR while the latter was mediated by ß-adrenergic and muscarinic receptors. CONCLUSION: Vasorelaxation effect by AESP and MESP involved nitric oxide pathway which is possibly mediated by the autonomic receptors. SUMMARY: This is the first study that reveals significant vasorelaxation effect induced by Syzygium polyanthum leaves extract. Vasorelaxation maybe one of the possible mechanisms for its ability to reduce blood pressure. This study also suggested that the vasorelaxation effect by this plant extract may involve nitric oxide pathway mediated by the autonomic receptors. Abbreviations Used: AESP: Aqueous extract of Syzygium polyanthum leaves. MESP: Methanolic extract of Syzygium polyanthum leaves. SHR: spontaneously hypertensive rat, WKY: Wistar-Kyoto rat.

Incorporation of dietary fibre-rich oyster mushroom (Pleurotus sajor-caju) powder improves postprandial glycaemic response by interfering with starch granule structure and starch digestibility of biscuit.

The purpose of this study was to determine the effects of Pleurotus sajor-caju (PSC) powder addition at 0, 4, 8 and 12% levels on the nutritional values, pasting properties, thermal characteristics, microstructure, in vitro starch digestibility, in vivo glycaemic index (GI) and sensorial properties of biscuits. Elevated incorporation levels of PSC powder increased the dietary fibre (DF) content and reduced the pasting viscosities and starch gelatinisation enthalpy value of biscuits. The addition of DF-rich PSC powder also interfered with the integrity of the starch granules by reducing the sizes and inducing the uneven spherical shapes of the starch granules, which, in turn, resulted in reduced starch susceptibility to digestive enzymes. The restriction starch hydrolysis rate markedly reduced the GI of biscuits. The incorporation of 8% PSC powder in biscuits (GI=49) could be an effective way of developing a nutritious and low-GI biscuit without jeopardizing its desirable sensorial properties.

Hypoglycemic and Antidiabetic Effect of Pleurotus sajor-caju Aqueous Extract in Normal and Streptozotocin-Induced Diabetic Rats.

INTRODUCTION: Pleurotus sajor-caju (PSC) is an edible oyster mushroom featuring high nutritional values and pharmacological properties. Objective. To investigate the hypoglycemic and antidiabetic effects of single and repeated oral administration of PSC aqueous extract in normal and diabetic rats. MATERIALS AND METHODS: A single dose of 500, 750, or 1000 mg/kg of the PSC extract was given to experimental rats to determine the effects on blood glucose (BG) and oral glucose tolerance test (OGTT). The effective dose (750 mg/kg) of PSC extract was repeatedly administrated daily for 21 days in diabetic rats to examine its antidiabetic effects in terms of BG control, body weight, urine sugar, HbA1c, and several serum profiles. RESULTS: The dose of 750 mg/kg showed the most significant BG reduction (23.5%) in normal rats 6 hours after administration in BG study (p < 0.05). In OGTT study, the same dose produced a maximum BG fall of 41.3% in normal rats and 36.5% in diabetic rats 3 hours after glucose administration. In 21-day study, treated diabetic rats showed significant improvement in terms of fasting BG, body weight, and urine sugar as compared to control diabetic rats. CONCLUSION: The study evidenced scientifically the beneficial use of PSC as an alternative medicine in diabetes management.