Nor Anita Megat Mohd Nordin

Piper sarmentosum inhibits ICAM-1 and Nox4 gene expression in oxidative stress-induced human umbilical vein endothelial cells

BackgroundAqueous extract of Piper sarmentosum (AEPS) is known to possess antioxidant and anti-atherosclerotic activities but the mechanism responsible for it remains unclear. In early part of atherosclerosis, nuclear factor-kappa B (NF-κB) induces the expression of cellular adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1), intracellular adhesion molecule-1 (ICAM-1) and E-selectin. NADPH oxidase 4 (Nox4) is the predominant source of superoxide in the endothelial cells whereas superoxide dismutase 1 (SOD1), catalase (CAT) and glutathione peroxidase (GPx) are the antioxidant enzymes responsible for inactivating reactive oxygen species. The present study aimed to investigate the effects of AEPS on the gene expression of NF-κB, VCAM-1, ICAM-1, E-selectin, Nox4, SOD1, CAT and GPx in cultured human umbilical vein endothelial cells (HUVECs).MethodsHUVECs were divided into four groups:- control; treatment with 180 μM hydrogen peroxide (H2O2); treatment with 150 μg/mL AEPS and concomitant treatment with AEPS and H2O2 for 24 hours. Total RNA was extracted from all the groups of HUVEC using TRI reagent. Subsequently, qPCR was carried out to determine the mRNA expression of NF-κB, VCAM-1, ICAM-1, E-selectin, Nox4, SOD1, CAT and GPx. The specificity of the reactions was verified using melting curve analysis and agarose gel electrophoresis.ResultsWhen stimulated with H2O2, HUVECs expressed higher level of ICAM-1 (1.3-fold) and Nox4 (1.2-fold) mRNA expression. However, AEPS treatment led to a reduction in the mRNA expression of ICAM-1 (p < 0.01) and Nox4 (p < 0.05) in the H2O2-induced HUVECs. AEPS also upregulated the mRNA expression of SOD1 (p < 0.05), CAT (p < 0.01) and GPx (p < 0.05) in oxidative stress-induced HUVECs. There was no significant change in the mRNA expression of VCAM-1 and E-selectin.ConclusionThe expressional suppression of ICAM-1 and Nox4 and induction of antioxidant enzymes might be an important component of the vascular protective effect of AEPS.

Piper sarmentosum increases nitric oxide production in oxidative stress: a study on human umbilical vein endothelial cells

OBJECTIVE: Nitric oxide produced by endothelial nitric oxide synthase (eNOS) possesses multiple anti-atherosclerotic properties. Hence, enhanced expression of eNOS and increased Nitric oxide levels may protect against the development of atherosclerosis. Piper sarmentosum is a tropical plant with antioxidant and anti-inflammatory activities. This study aimed to investigate the effects of Piper sarmentosum on the eNOS and Nitric oxide pathway in cultured human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were divided into four groups: control, treatment with 180 μM hydrogen peroxide (H2O2), treatment with 150 μg/mL aqueous extract of Piper sarmentosum, and concomitant treatment with aqueous extract of PS and H2O2 for 24 hours. Subsequently, HUVECs were harvested and eNOS mRNA expression was determined using qPCR. The eNOS protein level was measured using ELISA, and the eNOS activity and Nitric oxide level were determined by the Griess reaction. RESULTS: Human umbilical vein endothelial cells treated with aqueous extract of Piper sarmentosum showed a marked induction of Nitric oxide. Treatment with PS also resulted in increased eNOS mRNA expression, eNOS protein level and eNOS activity in HUVECs. CONCLUSION: Aqueous extract of Piper sarmentosum may improve endothelial function by promoting NO production in HUVECs.

Effects of Swedish Massage Therapy on Blood Pressure, Heart Rate, and Inflammatory Markers in Hypertensive Women

Swedish Massage Therapy (SMT) is known for its therapeutic relaxation effects. Hypertension is associated with stress and elevated endothelial inflammatory markers. This randomized control trial measured the effects of whole body SMT (massage group) or resting (control group) an hour weekly for four weeks on hypertensive women. Blood pressure (BP) and heart rate (HR) were measured before and after each intervention and endothelial inflammatory markers: vascular endothelial adhesion molecules 1 (VCAM-1) and intracellular adhesion molecules 1 (ICAM-1) were measured at baseline and after the last intervention. Massage group (n=8) showed significant systolic BP (SBP) reduction of 12 mmHg (P=0.01) and diastolic BP (DBP) reduction of 5 mmHg (P=0.01) after four sessions with no significant difference between groups. Reductions in HR were also seen in massage group after sessions 1, 3, and 4 with significant difference between groups. VCAM-1 showed significant reduction after four sessions: the massage group showed reduction of 998.05 ng/mL (P=0.03) and the control group of 375.70 ng/mL (P=0.01) with no significant differences between groups. There were no changes in ICAM-1. In conclusion, SMT or resting an hour weekly has effects on reducing BP, HR, and VCAM-1 in hypertensive women.

Flavonoids of Piper sarmentosum and its cytoprotective effects against oxidative stress

Abnormalities in endothelial cell structure and function may lead to diseases such as thrombosis and atherosclerosis. Oxidative stress plays an important role in the pathogenesis of various cardiovascular diseases including atherosclerosis. Previous studies have shown a relationship between a diet rich in flavonoid and a reduced incidence of cardiovascular diseases. Piper sarmentosum (PS) is a plant with high flavonoid content and it possesses antioxidant and anti-atherosclerotic activities. Therefore this study aimed to investigate the flavonoids present in aqueous extract of PS (AEPS) and its cytoprotective effects in oxidative stress-induced human umbilical vein endothelial cells (HUVEC). AEPS contained high total phenolic content (91.02 ± 0.02 mg QE/g DM) and total flavonoid content (48.57 ± 0.03 mg GAE/g DM). Screening using high performance liquid chromatography (HPLC) technique showed the presence of rutin and vitexin as the main flavonoids in AEPS. HUVEC were exposed to 180 µM H2O2 and treated with various concentrations of rutin or vitexin (10 to 400 µM) for 24 hours. Both rutin and vitexin at the concentration of 150-400 µM significantly increased the viability of H2O2-induced HUVEC as denoted by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Therefore rutin and vitexin as the main flavonoids present in PS may be involved in the protective effects of PS against oxidative stress.

The use of Piper sarmentosum leaves aqueous extract (Kadukmy™) as antihypertensive agent in spontaneous hypertensive rats

BackgroundThe National Health and Morbidity Survey in 2011 estimated that 35.1% (5.7 million) of Malaysian adults aged 18 and older suffer from hypertension. Hypertension is still treated by conventional medicine despite its exact aetiology being unknown. Studies showed that oxidative stress and low availability of nitric oxide (NO) causes an increase in vascular wall tension and increase blood pressure. Piper sarmentosum (PS) a traditional Malay herbal plant is well known for its high antioxidant content. Antioxidant is useful in improving cardiovascular diseases particularly hypertension. Thus, it is beneficial to determine the effect of PS leaves aqueous extract (Kadukmy™) on the blood pressure, NO level, oxidative stress markers and serum cholesterol level of the Spontaneous Hypertensive Rats (SHR).MethodsRats were devided into five groups consisting of three treatment groups and two control groups. Baseline blood investigations were done before and following commencement of treatment. Spontaneous hypertensive rats were treated for 28 consecutive days and the blood pressure was measured weekly.ResultsKadukmy™ administration showed a significant reduction in systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) (P < 0.05), increased serum NO level (P < 0.05), reduced serum malondialdehyde (MDA) level (P < 0.05) and reduction of serum total cholesterol level in groups treated with Kadukmy-1™.ConclusionsThe result of the present study revealed that Kadukmy™ exerts its antioxidant activity to reduce oxidative stress damage, increase NO production and able to reduce blood pressure and cholesterol level.

Does oral ingestion of Piper sarmentosum cause toxicity in experimental animals

The prevalence of diabetes mellitus has reached epidemic proportion in Malaysia and worldwide. Scientific studies have shown that herbal plant Piper sarmentosum exhibits an antidiabetic property. Despite the extensive usage and studies of this herb as alternative medicine, there is paucity of the literature on the safety information of this plant. Thus, the present study aimed to observe the subacute toxic effects of Piper sarmentosum aqueous extract (PSAE) on the haematological profile, liver, and kidney in rats. The extract was administered by oral gavage to 6 male and female Sprague Dawley rats in daily dose of 50 mg/kg, 300 mg/kg, and 2000 mg/kg for 28 consecutive days. The control group received normal saline. General behavior of the rats, adverse effects, and mortality were observed for 28 days. The haematological and biochemical parameters were determined at baseline and after the treatment. PSAE did not show abnormality on the body weight and gross observation of internal organs. The haematological, biochemical and histopathological profiles showed minimal changes and variation within normal clinical range except for significant increase in serum potassium level that suggests the need of regular monitoring. Nevertheless, these findings suggested that PSAE up to 2000 mg/kg/day did not show subacute toxicity in Sprague Dawley rats.

Carotid intima media thickness among young males with cardiovascular risk factors

Introduction: Atherosclerosis is a condition where there are fat, cells and connective tissue deposition in the arterial wall. This may lead to reduced blood flow to the vital organs such as heart (ischemic heart disease) and brain (stroke). There is increasing prevalence of ischemic heart disease (IHD) among the young population, especially males. Carotid intima media thickness (CIMT) of the common carotid artery is a novel marker of atherosclerosis. The aims of the current study were to compare these markers among young males with cardiovascular disease (CVD) risk factors with healthy subjects and to determine the associations between these markers with other CVD risk factors. Methods: One hundred and eighty one men aged 20 to 40 years old were recruited from areas around Klang Valley. These comprised those having one or more risk factors of CVD [hypertension, abdominal obesity, dyslipidemia, first degree family history (FH) of CVD and smoker, n=128] and without risk factors (n=53). Measurements included peripheral blood pressure (BP), heart rate (HR), body mass index (BMI), waist circumference (WC), total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein (LDL) and fasting blood sugar (FBS). Right and left CIMT were also measured via B mode ultrasound. Data were analyzed via SPSS version 15 and significance level was set at P<; 0.05. Results: The mean age of all the subjects was 26.47 ± 5.09 years old. Those with risk factors were older, had significantly higher BP, BMI, WC and lipid profiles. Nevertheless, most of these variables were within normal range. Other variables such as HR and FBS were comparable between the two groups. After adjustment for age, those with CV risk factors had significantly higher right CIMT (0.5600 ± 1417 vs 0.5267 ± 0733 mm, P<;0.05) but not left CIMT (0.5450 ± 0.1500 vs 0.5167 ± 1033 mm, P > 0.05) compared to healthy group. Both CIMTs correlate with other major CV risk factors. The predictors of CIMT were age, SBP and WC which contribute about 12% of the variance in CIMT. Conclusion: Young males with CV risk factors have increased RCIMT and this may be the early markers of CVD.

Improvement of Cardiorespiratory Fitness in Young men with Cardiovascular Risks Participating in Pedometer-based Workplace Programme

Lack of exercise among Malaysians and the increasing cardiovascular disease morbidity and mortality among young men are of concern. In response to the wide use of pedometers, daily targets of 10,000 steps/day have been recommended. In Malaysia, data on workplace exercise intervention is scarce. A programme was designed to enable subjects to increase their level of walking as part of their daily work routine and self-monitored by using pedometers. The aim of this study was to evaluate the improvement of cardiorespiratory fitness (CRF) through a pedometer based walking programme at the workplace. A total of 70 young men (20-40 years old) who were sedentary, achieving less than 5,000 steps/day in casual walking with 2 or more cardiovascular risk factors were recruited in IKBN Hulu Langat. Subjects were randomly assigned to a control (CG) (n=34; no change in walking) and pedometer group (PG) (n=36; minimum target: 8,000 steps/day). Blood lipid profile, anthropometric and CRF were measured at baseline and after 12 weeks. At post intervention, the CG step counts were similar (4983 ± 366 vs 5697 ± 407 steps/day). The PG increased step count from 4996 ± 805 to 10,128 ± 511 steps/day (P<0.001). Results for lipid and anthropometrics variables were significantly improved for time and group effect (p<0.001). In PG, the CRF was significantly increased for time and effect (p<0.01) for VO2 peak from 31.54 ± 9.66 to 40.15 ± 9.55(ml/kg/min but no change in CG (31.46 ± 6.15 vs 31.60 ± 8.99 ml/kg/min). The walking programme improved health status in terms of improving biophysical profiles and cardiorespiratory fitness.

Correlation between lipid profile and finger photoplethysmogram morphological properties among young men with cardiovascular risk: A preliminary result

Photoplethysmogram (PPG) is a non-invasive optical technique that is widely used in many cardiovascular risk assessment and monitoring applications. PPG is the measure of blood volume change recorded from finger, toe, ear lobe, forehead and other location based on the application requirements. PPG has been adopted as a potential non-invasive technique for lipid profiling among young men aged between 20 - 40 years old. A total of 76 subjects were recruited from a non-active lifestyle with less than five thousand steps a day in walking with cardiovascular risk which includes two or more cardiovascular risks factor. The parameter measurements that were taken in the study were the finger PPG signal recording and the cholesterol values which were the HDL, LDL and triglyceride (TG). The morphological analysis clearly differentiates the subjects with different combination of lipid profiles. In this study we are able to categorize subjects into four different groups: a) normal LDL: normal TG; b) high LDL: normal TG; c) normal LDL: high TG; d) high LDL: high TG. The identified morphological classification will need further analysis and automated classification to establish a non-invasive low cost optical based screening method for lipid profiling in future.