Effects of palm oil derived tocotrienol rich fraction (TRF) on coronary risk markers: in vitro and in vivo studies / Suhaila Abd Muid

Tocotrienol is one of the vitamin E compounds which have potent anti-oxidant activity leading to reduction in oxidative stress and inflammation. Oxidative stress and inflammation are now emerging as pivotal factors in the pathogenesis of atherosclerosis and coronary artery disease (CAD). However, optimal concentrations of palm oil derived tocotrienol rich fraction (TRF) that may lead to reduction of oxidative stress and inflammation in in vitro and in vivo studies are still unclear. The objectives of this study were to determine the optimal concentrations of TRF which leads to the highest antioxidant activity and reduction of inflammation markers in vitro and to study the effects of palm oil derived TRF vitamin E capsules (Palmvitee) contained low dose tocotrienol on oxidative stress in patients with non-familial hypercholesterolaemia (NFH). Antioxidant activities were assessed by Ferric thiocyanate (FTC), 1,1-diphenyl-2picrylhydrazyl (DPPH) radical scavenging activities and dichlorofluorescein diacetate (DCFHDA) assays. Supernatant of stimulated endothelial cells was measured for the production of inflammatory markers [e-selectin, p-selectin, soluble intercellular cell adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1) and inter leukin-6 (IL-6)]. Sixty-six patients with NFH (38 males, 28 females, age ± SD age = 46.9 ± 9.4 years) were recruited and randomised to 3 treatments arm, which were palmvitee 300 mg/day (NFHe), atorvastatin 10 mg/day plus placebo (NFHsp) or atorvastatin 10 mg/day plus palmvitee (NFHse). Fasting serum lipids (FSL), oxidised LDL (ox-LDL), malondialdehyde (MDA) and 8-epi-PGF2α were measured at baseline (BL) and 2 weeks, 12 weeks and 36 weeks post-randomisation. Normocholesterolaemic (NC) subjects were recruited in parallel. In the FTC assay, TRF at concentrations 10 and 100 µg/ml showed the highest percentage (%) of inhibition, 96.4 ± 0.2% and 96.3 ± 0.2% respectively. In the DPPH assay, optimal TRF concentration was observed at concentration 62.5 µg/ml with the highest % inhibition (85.2 ± 0.8%). Lowest % increase of DCF fluorescence production was shown by induced RAW 264.7 cells incubated with 88 µg/ml TRF concentration and it was significantly lower than DCF fluorescence produced by RAW 264.7 cells incubated with inducer (lipopolysaccharides and interferon gamma) alone , 95.0 ± 1.4% vs. 194 ± 18.5%, p<0.05. -The results from the DPPH and DCFHDA assay were consistent with the findings from the .FTC method which reported optimal TRF concentration within the range of 10-100 µg/ml, Optimal TRF concentrations leading to maximal inhibition of e-selectin , p-selectin, sICAM-1 , sVCAM-1 and IL-6 in stimulated endothelial cells were 0.8, 0.4, 0.8, 3.4 and 0.2 µg/ml, with percentage inhibition of 88.0 ±2.0%, 88.0 ±0.1%, 55.0 ±3.0%, 75.0 ±4.0% and 56.0 ±12.0%, respectively. NFHe group showed neutral effects on FSL, but reductions in MDA (p<0.005), ox-LDL (p<0.05) and 8-epi-PFGF2α (p<0.05) levels at 12 weeks compared to BL. NFHsp and NFHse groups showed similar reductions in 'Fe and LDL at 2 weeks (p<O.OOO 1) compared to BL. TO level was also reduced at 2 weeks in both NFHsp (p<O.O 1) and NFHse (p<0.005) groups compared to BL. NFHsp and NFHs

Modulation of inflammation and endothelial activation with spaceflight travel: tocotrienols as atheroprotective agents / Suhaila Abd Muid

The effects of immediate spaceflight travel on inflammation and endothelial activation in human endothelial cells (ECs) is not yet established. In addition, theexpression of these biomarkers in revived live ECs recovered from a spaceflight travel has not been reported so far. Endothelial activation is preventable. One of the major preventive strategies is the usage of antioxidants. Tocotrienols (TCTs) is a more potent antioxidant than tocopherol (TOC). However, the role of Tocotrienol enriched mixed fraction (TEMF) and pure TCT isomers as a potential potent antiatherosclerotic agent in human ECs compared to pure a-TOC is not well established.The anti-atherosclerotic mechanism of TCTs is also unclear. The objectives of this study were to investigate (i) the effects of spaceflight travel on the protein and gene expression of inflammation and endothelial activation, nuclear factor kappa B (NFkB) and endothelial nitric oxide synthase (eNOS) in human ECs compared to ground controls, (ii) the protein and gene expression of inflammation and endothelial activation, NFkB, signal transducer and activator of transcription-3 (STAT-3) and eNOS in revived live human ECs compared to matched controls (iii) the effects of TEMF, pure TCT isomers, and a-TOC on inflammation, endothelial activation, monocytes binding activity, NFkB and eNOS, and (iv) the most potent pure TCT isomers on the inhibition of the inflammation, endothelial activation, monocytes binding activity, NFkB and eNOS biomarkers in lipopolysaccharides (LPS) stimulated human ECs. The culture medium and ECs from post-spaceflight, revived and corresponding controls were collected and measured for protein and gene expression of cytokines (IL-6 and TNF-a), adhesion molecules (ICAM-1, VCAM-1 and e-selectin), NFkB and/or STAT-3 and eNOS. Human umbilical vein endothelial cells (HUVECs) were incubated with various concentrations of TEMF, pure TCT isomers and a-TOC (0.3-10 |iM) together with, lipopolysaccharides (LPS) for 16 hours. Culture medium and cells were collected and measured for the protein and gene expression of cytokines, adhesion molecules, NFkB and eNOS. The immediate post-spaceflight cells showed enhanced expression of cytokine (IL-6), adhesion molecules (ICAM-1 and VCAM-1) and NFkB compared to ground controls. Following post spaceflight, the revived cells were shown to have increased expression of IL-6 , ICAM-1 and STAT-3. TEMF and pure TCT isomers reduce IL-6, ICAM-1, VCAM-1, e-selectin, monocytes binding activity, NFkB and induce eNOS expression. Area under the analysis revealed that pure TCT, particularly y- and 8-isomers have better reduction of inflammation and endothelial activation and greater eNOS increment than TEMF. Delta (8)-TCT is the most potent TCT isomers in terms of as an atheroprotective agent. Spaceflight travel leads to enhanced inflammation and endothelial activation and these remain elevated even after 3 months post spaceflight travel. This study provided a better understanding on the modulation of inflammation and endothelial activation associated with space travel and may direct future studies in the prevention of atherosclerosis in space travel. TEMF and pure TCT isomers exhibit anti-atherosclerotic properties with great potential as atheroprotective agents. The possible pathway for its anti-atherosclerotic activity is through the NFkB deactivation. a-TOC has inhibitory effects on the antiatherosclerotic properties of TCTs in TEMF

Inflammation and Vascular Calcification Causing Effects of Oxidized HDL are Attenuated by Adiponectin in Human Vascular Smooth Muscle Cells

The role of oxidized high- density lipoprotein (oxHDL) and the protective effects of adiponectin in terms of vascular calcification is not well-established. This study was conducted to investigate the effects of oxHDL with regard to inflammation and vascular calcification and to determine the protective role of adiponectin in attenuating the detrimental effects of oxHDL. Cell viability, mineralization, and calcification assays were conducted to optimize the concentration of oxHDL. Then, human vascular smooth muscle cells (HAoVSMCs) were incubated with β-glycerophosphate, HDL, oxHDL, adiponectin, or the combination of oxHDL with adiponectin for 24 h. Protein expression of IL-6, TNF-α, osterix, RUNX2, ALP, type 1 collagen, osteopontin, osteocalcin, WNT-5a, NF-ĸβ(p65), cAMP and STAT-3 were measured by ELISA kits. OxHDL induced vascular calcification by promoting the formation of mineralization nodules and calcium deposits in HAoVSMCs. This was accompanied by an increased secretion of IL-6, osterix, WNT-5a and NF-ĸβ (p65). Interestingly, these detrimental effects of oxHDL were suppressed by adiponectin. Besides, incubation of adiponectin alone on HAoVSMCs showed a reduction of inflammatory cytokines, osteoblastic markers (RUNX2, osterix and osteopontin), WNT-5a and NF-ĸβ (p65). This study exhibits the ability of oxHDL in inducing inflammation and vascular calcification and these detrimental effects of oxHDL can be attenuated by adiponectin

Alpha-mangostin (Garcinia mangostana Linn.) and its potential application in mitigating chronic wound healing

Wound healing is a complex and dynamic cellular process to restore tissue function. Current treatments for chronic wounds especially diabetic ulcers are expensive, with adverse effects. Recently, numerous researchers have focused on the potential effect of natural products on wound healing. One of them is mangosteen (Garcinia mangostana Linn). It is a well-known tropical fruit that is native to Southeast Asia. The active ingredient of mangosteen pericarp contains xanthones that exhibit a wide range of pharmacological activities, including anti-inflammatory and anti-bacterial properties which are the core elements needed in wound healing. Firstly, this review discusses the concepts of abnormal and normal wound healing mechanisms. Then an in-depth observation of the pharmacological activities of mangosteen and its derivatives was presented to study their potentially beneficial applications in the treatment of chronic wound healing which is a contemporary medical issue

Antimycotic screening of 58 Malaysian plants against plant pathogens

Ethanolic extracts of 58 Malaysian plants belonging to 24 different families were screened for antifungal activity against seven plant pathogens using the filter paper disc diffusion technique. Two varieties of Piper betle, showed strong activity against all the pathogens tested (Colletotrichum capsici, Fusarium pallidoroseum, Botryodiplodia theobromae, Alternaria alternata, Penicillium citrinum, Phomopsis caricae-papayae and Aspergillus niger), with inhibition diameters significantly (P<0·01) bigger than 2·5 mg ml−1 prochloraz or 10 mg ml−1 clotrimazole. The minimum inhibitory concentrations of the ethanolic extracts of P. betle against these plant pathogens ranged between 0·01 mg ml−1 and 1 mg ml−1. Thirty-four other plants (Kucing gala, Limau batik, Bertholletia excelsa, Bixa orellana, Caesalpinia pulcherrima, Cerbera odollam (fruits and leaves), Colocasia gigantea, Curcuma domestica, Curcuma manga, Derris eliptica, Elephantopus scaber, Eleusine indica, Eugenia polyantha, Euphorbia hirta, Euphorbia tirucalli, Gardenia florida, Hedyotis auricularia, Hibiscus rosa-sinensis, Juniperus chinensis (three varieties), Lawsonia inermis, Lecythis ollaria, Mentha arvensis, Mimusops elengi, Ocimum sanctum, Phyllanthus niruri, Piper nigrum, Piperomia pellucida, Pedilanthus tithymaloides, Polygonum minus, Spondias dulcis, Solanum nigrum, Tinospora tuberculata) showed selective antifungal activity, while 21 species were inactive

Ficus deltoidea var. kunstleri extract administration in Hypercholesterolaemic, atherosclerotic rabbits: effects on organ function, morphology, and atherosclerosis development

Ficus deltoidea (FD) is used in traditional Malay medicine to treat various ailments and has been shown to be safe in toxicity studies. However, the information on the safety and efficacy of FD in the atherosclerosis-induced animal model is limited. This study aims to investigate the safety of FD var. kunstleri (FDK) extract on high cholesterol diet (HCD)-induced atherosclerotic rabbits and its efficacy in treating atherosclerosis. New Zealand White rabbits were randomly divided into two groups: G1 (1% HCD for 4 weeks) and G2 (1% HCD for 8 weeks). Each group was randomised into FDK700 (700 mg FDK/kg/day for G1 and G2), FDK800 (800 mg FDK/kg/day for G2), simvastatin (5 mg/kg/day) and placebo. The body weight, blood pressure, serum biochemistry and histopathological examination were obtained to assess any toxicity signs. Fasting lipid profile, soluble c-reactive protein (sCRP) level and atherosclerotic plaque formation were compared between treated and placebo groups to evaluate treatment efficacy. Results: No significant differences were observed in all safety parameters between the treated and placebo groups (p<0.05). FDK treatment did not show significant differences in all parameters evaluated in both treatment arms. In conclusion, FDK extract up to 800 mg/kg is safe for use in atherosclerotic rabbits. It has neutral effects on lipid profile, inflammation and atherosclerosis formation

HDL and its subpopulation (HDL2 AND HDL3) promote cholesterol transporters expression and attenuate inflammation in 3t3-l1 mature adipocytes induced by tumor necrosis factor-alpha

Obesity activates inflammation causing dysfunction of adipocytes. Increasing high-density lipoprotein (HDL) levels in obesity may be beneficial in overcoming this effect. However, not much data is available on the effects of HDL and its subpopulations in inflamed adipocytes. The objective of this study was to investigate the effects of total HDL (tHDL) and the comparison between its subpopulations (HDL2 & HDL3) on protein and gene expression of cholesterol transporters, inflammation, and adipokines in TNF-α stimulated 3T3-L1 mature adipocytes. TNFα alone had lower adiponectin and higher protein and gene expression of IL-6 and NF-ĸβ (p65) compared to unstimulated adipocytes and these effects were attenuated by HDLs especially HDL3 (in most of the biomarkers). HDL and its subpopulation had higher cholesterol transporters expression in 3T3-L1 mature adipocytes induced by TNF-α compared to unstimulated cells. Increment of cholesterol transporters expression by HDL leads to reduce secretion of inflammatory markers [IL-6 & NF-kB (p65)] and visfatin and increases adiponectin secretion in the inflamed mature adipocytes. HDL exhibits beyond its reverse cholesterol transporter property by exhibiting anti-inflammatory effects thru the deactivation of NF-ĸβ (p65). This may contribute to reducing the progression of obesity-related complications

HDL and its subpopulation (HDL2 and HDL3) promote cholesterol transporters expression and attenuate inflammation in 3T3-L1 mature adipocytes induced by tumor necrosis factor-alpha

Obesity activates inflammation causing dysfunction of adipocytes. Increasing high-density lipoprotein (HDL) levels in obesity may be beneficial in overcoming this effect. However, not much data is available on the effects of HDL and its subpopulations in inflamed adipocytes. The objective of this study was to investigate the effects of total HDL (tHDL) and the comparison between its subpopulations (HDL2 & HDL3) on protein and gene expression of cholesterol transporters, inflammation, and adipokines in TNF-α stimulated 3T3-L1 mature adipocytes. TNFα alone had lower adiponectin and higher protein and gene expression of IL-6 and NF-ĸβ (p65) compared to unstimulated adipocytes and these effects were attenuated by HDLs especially HDL3 (in most of the biomarkers). HDL and its subpopulation had higher cholesterol transporters expression in 3T3-L1 mature adipocytes induced by TNF-α compared to unstimulated cells. Increment of cholesterol transporters expression by HDL leads to reduce secretion of inflammatory markers [IL-6 & NF-kB (p65)] and visfatin and increases adiponectin secretion in the inflamed mature adipocytes. HDL exhibits beyond its reverse cholesterol transporter property by exhibiting anti-inflammatory effects thru the deactivation of NF-ĸβ (p65). This may contribute to reducing the progression of obesity-related complications

Urinary apolipoprotein A1 and its potential as a biomarker for coronary artery disease in young adults

INTRODUCTION: Very few studies have focused on exploring the utilisation of urinary protein biomarkers to improve the risk stratification of coronary artery disease (CAD) in young adults. Apolipoprotein A1 (ApoA1) as a primary constituent protein of Highdensity Lipoprotein (HDL) known to modulate cholesterol metabolism exhibits promising properties to be used as a protein biomarker, specifically for CAD in young adults. Thus, this study is aimed to evaluate the potential of urinary ApoA1 as a urinary biomarker of CAD in young patients with acute myocardial infarction (AMI). MATERIALS AND METHOD: This case-control study recruited 40 newly diagnosed AMI patients and 40 healthy control subjects aged 18–45. Urine samples were collected from all subjects. Once centrifuged, the supernatant was collected and stored at -80 °C until further analysis. The urinary concentration of ApoA1 was quantified using the ApoA1 Enzyme-linked Immunosorbent Assay (ELISA) kit according to the manufacturer's protocol. All subjects' risk factors were determined and documented, such as smoking status, Body Mass Index (BMI), blood pressure, plasma total cholesterol, and glucose levels. RESULTS: The mean age of AMI patients was higher than the controls; 37.1 1 ± 5.2 and 31.6 ± 8.1 years respectively. The mean urinary concentration of ApoA1 of AMI patients was significantly higher than the controls (12. 442 ± 3.571 vs. 10.067 ± 5.606 ng/ mL (p0.05). CONCLUSION: A significant elevation of urinary excretion of ApoA1 in AMI young adults demonstrated its potential use as a urinary protein biomarker for CAD in young adults

In Vitro Atheroprotective Effects of Trigonella Foenum Graecum (TFG) and its Saponins in LPS-Stimulated Human Coronary Artery Endothelial Cells

There has been a shift towards utilizing natural products as an adjunct therapy to standard treatment in the prevention of coronary artery disease, and Trigonella foenum graecum (TFG) is one of the potential natural products of interest. In the present study, we attempted to determine the effects of TFG and its saponins on atherosclerosis related biomarkers in vitro. Protein expression of markers of inflammation, endothelial activation and transcription factors were measured by Procarta™ and ELISA assays. Gene expression of the same markers were determined by qPCR and the interaction between monocytes and HCAECs were evaluated through monocyte binding assay following 16 h of treatment with TFG and saponins. Both TFG and its saponins exhibited reducing effects on atherosclerosis-related markers. Based on the area under the curve (AUC) analysis, TFG reduced protein and gene expressions of ICAM-1 and VCAM-1 better than the saponins, while saponins reduced E-selectin expression better than TFG. Saponins showed a reduction of gene and protein expressions of IL-6, IL-8, NF-κB p50 and p65 better than TFG. TFG is more effective in reducing binding of monocytes to endothelial cells than saponins. TFG better reduced endothelial activation but exerted weaker anti-inflammatory effects than saponins, suggesting the possible synergism with other compounds in the crude extract which enhances attenuation of endothelial activation while inhibiting anti-inflammatory properties of saponins in the crude extract