Research progress on approaches for detection of vitamin E in human blood / 预防医学

@#There are eight forms of vitamin E in human blood, including α-, β-, γ-, δ-tocopherols and α-, β-, γ-, δ-tocotrienols. As the most abundant and active form of vitamin E, α-tocopherol is widely accepted as a reliable indicator for nutritional assessment of body vitamin E status across the world. Considering that different vitamin E forms have diverse biological activities, separation and detection of different vitamin E forms in human blood facilitates the understanding of the association between vitamin E and diseases. In this review, the advances in sample-pretreatment techniques and detection techniques for vitamin E in human blood were presented. Currently, the sample-pretreatment techniques include solid-phase extraction, liquid-liquid extraction, dispersive liquid-phase microextraction, supported liquid extraction and direct protein precipitation; the detection techniques include automatic biochemical analysis, enzyme-linked immunosorbent assay, gas chromatography, liquid chromatography and ultra-high performance supercritical fluid chromatography mass spectrometry. This review summarizes the characteristics and scope of above-mentioned techniques used for detection of vitamin E in human blood, so as to provide insights into the selection of an appropriate method for inspection technicians.

Effect of Tocotrienol Rich Fraction (TRF) On Muscles Reinnervation After Sciatic Nerve Crush Injury In Rats

@#Introduction: Muscle denervation is a process where muscles lose nerve supply due to neural damage and this may lead to paralysis in human. Muscle denervation is mainly caused by peripheral nerve injuries especially in the lower extremities that resulted in devastating effect on human daily functions and routines. Tocotrienol Rich Fraction (TRF) consist of 75% of tocotrienols have shown potential neuroprotective properties. The objective of this study is to observe motor coordination and histological characteristics on muscles that underwent sciatic nerve crush injury and supplemented with TRF. Methods: A total of 104 Sprague-Dawley rats were divided into four groups; normal group (n=8) with no sciatic nerve crush injury, negative control (n=32) with sciatic nerve crush injury at hindlimb without treatment, positive control (n=32) sciatic nerve crush injury treated with 500 µg/kg/day of methylcobalamin, and experimental group (n=32) of rats that underwent sciatic nerve crush injury and treated with 200 mg/kg/day of TRF. Result: Skeletal muscles which located at hind limb; Soleus Muscle and Extenstor Digitorum Longus Muscle (EDL) muscle have shown an increasing in weight when it is supplemented with TRF 200 mg/kg/day and improved myelin layer of nerve. Conclusion: This study showed that TRF has the potency to improve reinnervation rate and neuron supply in hind muscle.

The Effect of Tocotrienol-Rich Fraction Supplementation on the Ovarian Metabolome and the Quality of Oocyte in Aging Mice

@#Ovarian aging has been associated with oxidative stress and loss of ovarian function. Tocotrienol has been proven to exert beneficial effects on the female reproductive system. However, the role of tocotrienol in affecting metabolism in the ovary and subsequently improving the quality of oocytes in aging mice remains unknown. Therefore, the relationship between metabolic changes in the ovary and the quality of oocytes in aging mice following tocotrienol-rich fraction (TRF) supplementation was investigated. Six-week-old female mice were used as the Young group. Six-month-old aged female mice were divided into four groups; the first group was given tocopherol-stripped corn oil (vehicle control) while the other three groups were supplemented with TRF at the dose of 90, 120, and 150 mg/kg, respectively. The treatment was given orally for two months. At the end of the treatment, mice from all groups were superovulated and then euthanised. Oocyte quality was assessed and non-targeted metabolomic analysis of the ovarian tissues was performed using liquid chromatography-tandem mass spectrometry of quadrupole time-of-flight (LC-MS Q-TOF). Percentages of normal oocytes were higher (p<0.001) while abnormal oocytes were lower (p<0.001) in TRF 150 mg/kg group compared to that of the control. Seventeen metabolites were identified to be significantly different in the ovarian tissue of the aging group when compared to the young group. 14 metabolites were identified to be significantly different in the ovarian tissue between the control and TRF supplemented groups. Pathway analysis revealed significantly altered metabolic pathways for fatty acid and amino acid metabolism that might influence the quality of oocytes. In conclusion, TRF supplementation causes metabolic changes in the ovary that delay the consequences of aging, thus improving the quality of oocytes in aging mice.

Tocotrienol-Rich Fraction (TRF) Improves the Viability of Wild-Type Saccharomyces cerevisiae in the Initial Stationary Phase

@#Palm oil tocotrienol-rich fraction (TRF) containing majorly of αα, β, γ andδδ-tocotrienols and some αα-tocopherols, was reported to have anti-ageing effects in both human and non-human model organisms, but still remains unexplored in the yeast model. It was reported to have the ability to extend the lifespan of several organisms. Chronological lifespan is one of the means to measure ageing in yeasts. The effect of TRF on the viability of three strains of Saccharomyces cerevisiae (wildtype, CTT1ΔΔand GPx2Δ) was studied. Phenotypic growth analysis of all strains was carried out for 15 hours by measuring the absorbance at OD600nm and cell counting. The optimum dose of TRF was optimised by determining the number of colony-forming unit by the wild-type strain at the end of a 24-hour treatment with TRF (ranging from 0μg/ml to 300μg/ml). TRF at 300 μg/mL showed the best result, and selected as a working dose. Treatment of cells with 300 μg/mL of TRF improved the viability of the wild-type strain in the initial stationary phase, but not on the knockout strains. These finding suggests that TRF has a potential in prolonging the chronological lifespan of S. cerevisiae, and perhaps other organisms as well

Vitamin E: An Antioxidant With Anticancer Properties?

@#More studies are now focusing on vitamin E as an anticancer agent for its good effects in many in-vitro studies. Current studies proposed that vitamin E might be a suitable candidate as an alternative treatment for cancer due to its antioxidant properties. Vitamin E act as an antioxidant by their long-chain polyunsaturated fatty acids, and thus the integrity of membranes in the cells is maintained and consequently retain the bioactivity of the cells. This mini review will focus on the activity of vitamin E as an antioxidant to protect against cancer in in-vitro, in-vivo, and clinical studies. Although most studies reported great outcomes for the anticancer activity of vitamin E, there were some conflicting data. To date, studies on effects of vitamin E are still undergoing where researchers are still debating on the positive and negative effects of vitamin E as an anticancer therapeutic action.

Chemical constituents from Garcinia multiflora fruits / 中草药

Objective To study the chemical constituents of the fruits of Garcinia multiflora. Methods The constituents were isolated and purified by silica gel column chromatography and semipreparative HPLC, and their structures were elucidated by means of spectroscopic analyses. Results Six compounds were obtained and identified as multiflorone A (1), sargaol (2), δ-tocotrienol (3), luteolin (4), 1,3,6,7-tetrahydroxyxanthone (5), terephthalic acid bis (2-ethyl-hexyl) ester (6). Conclusion Compound 1 is a new homoadamantyl type phloroglucinol, and compounds 2, 3 and 6 are isolated from this plant for the first time.

The Effects of Annatto Tocotrienol on Body Composition and Serum Adiponectin, Leptin and Glucose Level in a Rat Model of Androgen Deficiency Induced by Buserelin

@#Androgen ablation therapy using gonadotropin-releasing hormone agonists is reported to be associated with metabolic abnormalities. Annatto tocotrienol (AnTT) is reported to reduce the expression of genes related to adipogenesis but the mechanism remains elusive. This study sought to determine the effects of annatto tocotrienol on body composition (lean and fat mass), serum adiponectin, leptin, and glucose levels in male rats treated with buserelin, a testosterone ablation agent. Three-month-old male Sprague Dawley rats (n=32) were randomly divided into four groups. The normal control (n=8) was given corn oil orally daily and normal saline subcutaneously daily. The remaining groups were injected with buserelin subcutaneously (75µg/kg/day). The buserelin group (n=8) was given corn oil orally, while the treatment groups were supplemented orally with AnTT at 60 or 100 mg/kg (n = 8/group). After treatment of 12 weeks, rats were euthanized. Dual-energy x-ray absorptiometry was performed to determine the lean and fat mass of the rats. Blood was collected for the evaluation of adiponectin, leptin and glucose levels. After 12 weeks, the lean mass, fat mass, adiponectin and leptin levels for all groups increased significantly compared to their respective baseline levels irrespective of their treatment (P<0.05). All groups, except rats receiving AnTT at 60 mg/kg, experienced a significant increase in glucose level after 3 months (P<0.05). Androgen ablation and AnTT do not influence body composition, adiponectin and leptin levels in male rats. However, annatto tocotrienol at 60 mg/kg may improve glucose metabolism.

Comparing the effects of vitamin E tocotrienol-rich fraction supplementation and α-tocopherol supplementation on gene expression in healthy older adults

OBJECTIVES This study aims to compare the differential gene expression resulting from tocotrienol-rich fraction and α-tocopherol supplementation in healthy older adults. METHODS A total of 71 eligible subjects aged 50 to 55 years from Gombak and Kuala Lumpur, Malaysia, were divided into three groups and supplemented with placebo (n=23), α-tocopherol (n=24) or tocotrienol-rich fraction (n=24). Blood samples were collected at baseline and at 3 and 6 months of supplementation for microarray analysis. RESULTS The number of genes altered by α-tocopherol was higher after 6 months (1,410) than after 3 months (273) of supplementation. α-Tocopherol altered the expression of more genes in males (952) than in females (731). Similarly, tocotrienol-rich fraction modulated the expression of more genes after 6 months (1,084) than after 3 months (596) and affected more genes in males (899) than in females (781). α-Tocopherol supplementation modulated pathways involving the response to stress and stimuli, the immune response, the response to hypoxia and bacteria, the metabolism of toxins and xenobiotics, mitosis, and synaptic transmission as well as activated the mitogen-activated protein kinase and complement pathways after 6 months. However, tocotrienol-rich fraction supplementation affected pathways such as the signal transduction, apoptosis, nuclear factor kappa B kinase, cascade extracellular signal-regulated kinase-1 and extracellular signal-regulated kinase-2, immune response, response to drug, cell adhesion, multicellular organismal development and G protein signaling pathways. CONCLUSION Supplementation with either α-tocopherol or tocotrienol-rich fraction affected the immune and drug response and the cell adhesion and signal transduction pathways but modulated other pathways differently after 6 months of supplementation, with sex-specific responses.

The Effectiveness of Tocotrienol Rich Fraction and Αlpha Tocoferol with Combination of Vitamin C in The Management of Systemic Inflammatory Response Syndrome (SIRS)

The pathophysiology of systemic inflammatory response syndrome (SIRS) had been described to involve various strong oxidative reactions affecting the status and progress of the patients. Antioxidant therapy had been suggested in many studies involving SIRS management. The objective of this study was to compare the role of vitamin E Tocotrienol and vitamin E Tocopherol combined with vitamin C as antioxidant therapy in the management of critically ill patients diagnosed with SIRS, admitted to the intensive care unit and high dependency wards of Universiti Kebangsaan Malaysia Medical Centre (UKMMC). It was a single blind randomized clinical trial with a total of 72 patients in which 44.4% Malays, 34.7% Chinese, 19.4% Indians and 1.4% others with 59.7% males and 40.3% females were recruited. Patients in TRI E group received Tocotrienol with Vitamin C while TOCO group received Tocopherol with Vitamin C and a control group did not receive any antioxidant. The clinical parameters (heart rate, respiratory rate, systolic blood pressure) showed improvements with significant difference at the end of study (post-intervention) as compared to admission (pre-intervention).Whereas, the sepsis (temperature, PCT, CRP and WBC) and oxidative stress (8-OHdG/Creatinine) parameters showed improvements with significant difference at the end of study (post-intervention) as compared to admission (pre-intervention). The TRI E group showed obvious improvement in clinical, sepsis and oxidative stress parameters, as compared to TOCO and control groups. This study showed that Vitamin E Tocotrienol and Vitamin E Tocopherol in combination with Vitamin C demonstrated significant improvement in the clinical and laboratory parameters during the management of SIRS. Therefore, Vitamin E in combination with Vitamin C had therapeutic benefits in the treatment of critically ill patients with SIRS.

Tocotrienol-rich Palm Oil Extract Induces NAD(P)H:quinone Oxidoreductase 1 (NQO1) Expression in Mice Liver.

A diet rich in tocotrienols has been shown to be beneficial for health. However, its detailed mechanism of action is still not fully understood. NAD(P)H:quinone oxidoreductase 1 (NQO1) is important in cellular defence due to its ability to detoxify reactive quinones and quinoneimines to their less toxic hydroquinones forms. The objective of this study is to investigate the effects of different doses of palm oil-derived tocotrienol rich fraction (palm TRF) supplementation on NQO1 gene and protein expression in mice livers. Western blot and qPCR assays were used to detect NQO1 expression levels. It was found that palm TRF significantly induced NQO1 expression at all doses given. In conclusion, palm TRF treatment increased NQO1 gene and protein expression in mice liver dose dependently, with the highest expression seen in mice treated with 1000 mg/kg palm TRF, followed by 500 and 200 mg/kg respectively.

Effects of adding tocotrienol-tocopherol mixed fraction and vitamin C supplementation on coronary risk biomarkers in patients with hypercholesterolaemia with moderate coronary risk.

This study was a prospective clinical trial to investigate the effects of adding combined tocotrienol-tocopherol mixed fraction (TTMF) and vitamin C (TTMF+C) supplementation on coronary biomarkers in non-statin and statin treated patients with hypercholesterolaemia (HC) with moderate coronary risk. A total of 35 patients were randomised at baseline into one of two groups, (G1) TTMF+C (320mg TTMF plus 500mg vitamin C) alone daily and (G2) TTMF+C (320mg TTMF plus 500mg vitamin C) plus atorvastatin 10 mg daily. The entire supplementation were taken for 12 months. Fasting serum samples were taken at baseline, 2weeks, 3months, 6months and 12months post-randomisation and analysed for inflammatory biomarkers; high sensitivity C- reactive protein (hsCRP) and interleukin-6 (IL6). Combination of TTMF and vitamin C supplementation leads to neutral effects on lipid profiles and inflammation; with no added benefit in statin-treated HC patients with moderate coronary risk. This neutral effects may be attributed to the tocopherol composition in TTMF which could possibly attenuate any potential beneficial effects of tocotrienols. Clinical studies using pure tocotrienols in the absence of tocopherols would further confirm this.

Oral Administration of Tocotrienol Ameliorates Lead-Induced Toxicity in the Rat Brain

The occurrence of severe lead (Pb) poisoning has risen in certain countries. There is increasing evidence that chronic lead exposure disturbs the prooxidant: antioxidant balance in the brain tissue and alters brain histology. The present study observed the antioxidant effect of tocotrienol-rich fraction (TRF) on brain tissues of the experimental rats following lead poisoning. Eighteen (n=18) male Sprague-Dawley rats, 6-weeks old, were randomly divided into control (CTRL) group and experimental groups; fed with 0.2% w/v lead acetate, as PB2 group; and fed with 0.2% w/v lead acetate and daily TRF supplementation (200 mg/kg body weight) as PB2T group. The experiment was conducted for 30 days. At the end of the study, the brain tissues were harvested and histopathological changes of the hippocampal region were observed. Biochemical findings such as brain lead, TRF and malondialdehyde (MDA) levels, and erythrocyte superoxide dismutase (SOD) activity were determined. It was observed that atypical apoptotic-like and disorganized neurons were present in the hippocampal region of the untreated PB2 group compared to PB2T group. Biochemical parameters showed a significant decrease (p 0.05) was obtained for MDA level, there was a significant increase (p < 0.05) in the erythrocyte SOD activity in PB2T compared to PB2 and CTRL. Supplementation with TRF improved histopathological changes in the brain tissues caused by lead exposure in drinking water by reducing lead accumulation in the brain of experimental rats.

Eficacia de un complemento alimenticio con serenoa serrulata y tocotrienol-tocoferol frente a la alopecia androgenética y el efluvio telógeno femeninos: A propósito de un estudio piloto / Effectiveness of a dietary supplement with serenoa serrulata and tocotrienol-tocopherol against female androgenetic alopecia and telogen effluvium: Report of a pilot study

Introducción: la alopecia androgenética femenina (FAGA) y el efluvio telógeno (ET), son causas comunes de pérdida de cabello en las mujeres. Un hecho conocido en estos tipos de alopecia es la mayor actividad de la 5α-reductasa, que reduce la testosterona a dihidro-testosterona, andrógeno que acelera el ciclo capilar y acorta su duración. La Serenoa serrulata es una especie vegetal, cuyos componentes presentan un efecto inhibidor de la 5α-reductasa. Objetivo: el objetivo principal de este estudio, consistió en valorar la eficacia y la tolerancia, frente a la alopecia femenina de la administración diaria oral, de un complemento alimenticio con extracto de Serenoa serrulata, L-cistina, complejo tocotrienol-tocoferol, hierro, zinc y vitaminas H, B1, B2, B3, B5, B6 y B12 durante seis meses. Material y Método: estudio piloto unicéntrico, abierto, no aleatorizado, a simple ciego, de 180 días de duración, llevado a cabo en voluntarias con FAGA o ET. Se valoró el aspecto macroscópico del cabello, la densidad capilar mediante microfotografía, la relación anágena / telógena (A/T) mediante tricograma, el número de cabellos desprendidos mediante ensayos de peinado ("combing test") y de lavado ("wash test"). Se realizó una encuesta de autovaloración de los resultados. Resultados: la edad media de las participantes (n=10) fue de 40±15,5 años. El tiempo de evolución fue de 4,5±2 años y de tres meses para la FAGA y el ET, respectivamente. Al final del tratamiento las participantes presentaron un incremento medio del 6,9%, en la proporción de cabellos en fase anágena y la relación A/T media aumentó a 3,3. El 80% mostraron una mayor densidad capilar al finalizar el estudio. Al cabo de tres meses, todas las participantes presentaron una reducción media de la pérdida de cabellos del 56%, respecto de la visita basal. El 90% percibieron una mayor o igual cantidad de cabello al final del estudio y el 80% lo notaron más fuerte. No se registró ninguna molestia inducida por el tratamiento. Conclusiones: la buena evolución de las variables analizadas apunta a un beneficio terapéutico, frente a la FAGA y el ET. Estudios con mayor número de pacientes, son necesarios a futuro.

Introduction: female androgenetic alopecia (FAGA) and telogen effluvium (TE) are common causes of hair loss in women. A known fact in these types of alopecia is the increased activity of 5a-reductase which reduces testosterone to dihydrotestosterone, accelerating hair cycle and shortening its duration. Serenoa serrulata is a plant species whose components have an inhibitory effect on 5a reductase. Objective: the main objective of this study was to evaluate the efficacy and tolerance against female alopecia of a food supplement with Serenoa serrulata extract, L-cystine, tocotrienol-tocopherol complex, iron, zinc, and vitamins H, B1, B2, B3, B5, B6 and B12 administered daily for six months. Material and Methods: single-center, open-label, non-randomized, single-blind, 180-days pilot study conducted in women with FAGA or TE. The study assessed the macroscopic appearance of the hair, hair density by microphotograph, the anagen / telogen ratio (A/T) by trichogram, and the number of detached hair by combing and wash tests. Self-assessment survey of the results was performed. Results: the mean age of participants (n = 10) was 40 ± 15.5 years. FAGA and ET had an evolution of 4.5 ± two years and three months, respectively. After treatment the participants had an average increase of 6.9% in the proportion of hairs in anagen phase, and the mean A/T ratio increased to 3.3. Eighty percent or patients showed an increased capillary density at study end. After three months, all participants had a mean reduction of hair loss of 56% vs baseline. Ninety percent perceived a greater or equal amount of hair at the end of the study and 80% noticed it stronger. No adverse effects induced by treatment were observed. Conclusions: the good results of the analyzed variables suggest a therapeutic benefit against FAGA and ET. Additional research is warranted to confirm the observed data.

Dose-dependent Modulation of Lipid Parameters, Cytokines and RNA by -tocotrienol in Hypercholesterolemic Subjects Restricted to AHA Step-1 Diet.

Aims: Evaluate the consumption of -tocotrienol (free from tocopherols) on serum lipid parameters, and several cytokines (TNF-, IL-4, IL-6, IL-8, IL-10), including gene expression and circulating microRNAs (miRNAs) in hypercholesterolemic subjects. Study Design: The present preliminary dose-response study consisted of six phases. All hypercholesterolemic subjects took increasing doses of -tocotrienol (125, 250, 500,750 mg/d) plus AHA Step-1 diet for 4 weeks during the 30 weeks study period. Methodology: Hypercholesterolemic (n = 31; serum cholesterol > 5.2 mmol/L) subjects (males- 26/females 5; age range 50-71 years) were enrolled in the study from Wah Cantonment, Pakistan. Serum lipid parameters were measured by auto analyzers. Various plasma cytokines, cDNA, and miRNAs were estimated by Signosis kits. Results: All participants (n = 31) completed all phases of study. The -tocotrienol plus AHA Step-1 diet caused reductions in lipid parameters in a dose-dependent manner with maximum effects on serum total cholesterol (15%), LDL-cholesterol (18%), triglycerides (14%) with 250 mg/d dose (P< 0.001). Doses above 500 mg/d resulted ininduction in levels of all lipid parameters, except HDLcholesterol. The cytokines associated with cardiovascular disease (plasma TNF-, IL-2, IL-4, IL-6, IL-8, IL-10) were all down-regulated 39%-64% by -tocotrienol treatment (P< 0.01). Similar results were obtained with gene expression of these cytokines using whole blood messenger-RNA. In contrast, circulating miRNA-7a, miRNA-15a, miRNA-20a (anti-angiogenic), miRNA-21, miRNA- 29a, miRNA-92a, miRNA-200, miRNA-206 (skeletal muscle regeneration) down-regulated in hypercholesterolemic subjects, were up-regulated by -tocotrienol treatment as compared to baseline (P< 0.01). Conclusion: The present results confirm that consumption of -tocotrienol plus AHA Step-1 diet causes significant reduction in serum lipid parameters and several cytokines (TNF-, IL-2, IL-4, IL- 6, IL-8, IL-10) at a low optimal dose (250 mg/d). The capacity of -tocotrienol to modulate inflammation is partly attributable to dose-dependent properties of inhibition/activation, which may play a major role in future treatment of cardiovascular diseases.

Tocotrienol supplementation in postmenopausal osteoporosis: evidence from a laboratory study

OBJECTIVE: Accelerated bone loss that occurs in postmenopausal women has been linked to oxidative stress and increased free radicals. We propose the use of antioxidants to prevent and reverse postmenopausal osteoporosis. This study aimed to examine the effects of tocotrienol, a vitamin E analog, on bone loss due to estrogen deficiency. Our previous study showed that tocotrienol increased the trabecular bone volume and trabecular number in ovariectomized rats. In the current study, we investigated the effects of tocotrienol supplementation on various biochemical parameters in a postmenopausal osteoporosis rat model. MATERIALS AND METHODS: A total of 32 female Wistar rats were randomly divided into four groups. The baseline group was sacrificed at the start of the study, and another group was sham operated. The remaining rats were ovariectomized and either given olive oil as a vehicle or treated with tocotrienol at a dose of 60 mg/kg body weight. After four weeks of treatment, blood was withdrawn for the measurement of interleukin-1 (IL1) and interleukin-6 (IL6) (bone resorbing cytokines), serum osteocalcin (a bone formation marker) and pyridinoline (a bone resorption marker). RESULTS: Tocotrienol supplementation in ovariectomized rats significantly reduced the levels of osteocalcin, IL1 and IL6. However, it did not alter the serum pyridinoline level. CONCLUSION: Tocotrienol prevented osteoporotic bone loss by reducing the high bone turnover rate associated with estrogen deficiency. Therefore, tocotrienol has the potential to be used as an anti-osteoporotic agent in postmenopausal women. .

Effect of the tocotrienol-rich fraction on the lifespan and oxidative biomarkers in Caenorhabditis elegans under oxidative stress

OBJECTIVE: This study was performed to determine the effect of the tocotrienol-rich fraction on the lifespan and oxidative status of C. elegans under oxidative stress. METHOD: Lifespan was determined by counting the number of surviving nematodes daily under a dissecting microscope after treatment with hydrogen peroxide and the tocotrienol-rich fraction. The evaluated oxidative markers included lipofuscin, which was measured using a fluorescent microscope, and protein carbonyl and 8-hydroxy-2′-deoxyguanosine, which were measured using commercially available kits. RESULTS: Hydrogen peroxide-induced oxidative stress significantly decreased the mean lifespan of C. elegans, which was restored to that of the control by the tocotrienol-rich fraction when administered before or both before and after the hydrogen peroxide. The accumulation of the age marker lipofuscin, which increased with hydrogen peroxide exposure, was decreased with upon treatment with the tocotrienol-rich fraction (p<0.05). The level of 8-hydroxy-2′-deoxyguanosine significantly increased in the hydrogen peroxide-induced group relative to the control. Treatment with the tocotrienol-rich fraction before or after hydrogen peroxide induction also increased the level of 8-hydroxy-2′-deoxyguanosine relative to the control. However, neither hydrogen peroxide nor the tocotrienol-rich fraction treatment affected the protein carbonyl content of the nematodes. CONCLUSION: The tocotrienol-rich fraction restored the lifespan of oxidative stress-induced C. elegans and reduced the accumulation of lipofuscin but did not affect protein damage. In addition, DNA oxidation was increased. .

Evaluation of the protective effects of tocotrienol-rich fraction from palm oil on the dentate gyrus following chronic restraint stress in rats

Exposure to chronic restraint stress has been shown to cause a number of morphological changes in the hippocampal formation of rats. Tocotrienol, an isoform of vitamin E, exhibits numerous health benefits, different from those of tocopherol. Recent studies have demonstrated that tocotrienol prevents stress-induced changes in the gastric mucosa, thus indicating that it may also protect other organs such as the brain from the damaging effects of stress. Therefore, the aim of the present study was to investigate the protective effect of tocotrienol-rich fraction (TRF) extracted from palm oil on the dentate gyrus of rats following exposure to chronic restraint stress. Thirty-six male Sprague Dawley rats were divided into four groups: control, stress, tocotrienol and combination of stress and tocotrienol. Animals were stressed by restraining them for 5 hours every day for 21 consecutive days. TRF was administered via oral gavage at a dose of 200 mg/kg body weight. Our results showed that the plasma corticosterone level was significantly increased in response to stress, compared to the control. The results confirmed previous findings that chronic restraint stress suppresses cellular proliferation and reduces granule cell number in the dentate gyrus. However, TRF supplementation failed to prevent or minimize these stress-induced changes. Therefore, we conclude that TRF at the current dosage is not effective in preventing the morphological changes in the dentate gyrus induced by chronic restraint stress.

A exposição crônica ao estresse por restrição causa série de alterações morfológicas na formação do hipocampo de ratos. Tocotrienol, uma isoforma da vitamina E, apresenta inúmeros benefícios para a saúde, diferente do tocoferol. Estudos recentes demonstraram que o tocotrienol impediu alterações induzidas por estresse na mucosa gástrica, indicando, assim, a possibilidade de que ele pode, também, proteger outros órgãos, como o cérebro, dos efeitos nocivos do estresse. Dessa forma, o objetivo do presente estudo foi investigar o efeito protetor da fração rica em tocotrienol (TRF), extraída do óleo de palma, no giro denteado após exposição crônica ao estresse por restrição. Trinta e seis ratos machos Sprague Dawley foram divididos em quatro grupos: controle, estresse, tocotrienol e combinação de estresse e tocotrienol. Os animais foram estressados por restrição, 5 horas por dia, durante 21 dias consecutivos. TRF foi administrado por gavagem oral na dose de 200 mg/kg de peso corporal. Nossos resultados mostraram que o nível de corticosterona plasmática foi significativamente aumentado em resposta ao estresse em comparação ao controle. Os resultados confirmam os achados anteriores de que o estresse por restrição suprime a proliferação celular e reduz o número de células granulares do giro denteado. No entanto, a suplementação de TRF foi ineficaz para evitar ou minimizar as alterações induzidas por estresse. Assim, concluímos que TRF na dose corrente não é efetiva para prevenir as alterações morfológicas no giro denteado induzida por estresse crônico por restrição.

Identificação de δ tocotrienol e de ácidos graxos no óleo fixo de urucum (Bixa orellana Linné) / Identification of δ tocotrienols and fatty acids in fixed oil of annatto (Bixa orellana Linné)

Bixa orellana é uma planta nativa do Brasil, conhecida popularmente como urucum. O objetivo desse estudo foi extrair o óleo das sementes de B. orellana e, a partir dele, isolar e identificar o tocotrienol e quantificar seus ácidos graxos. A extração do óleo realizou-se em aparelho Soxhlet utilizando hexano como solvente extrator. O tocotrienol foi isolado por meio de métodos cromatográficos e identificado por espectrometria de ressonância magnética nuclear RMN 1H e RMN 13C. Os ácidos graxos foram quantificados por cromatografia gasosa acoplada ao espectrômetro de massa. Os resultados demonstraram a presença de δ -tocotrienol e, dentre os ácidos graxos, a ocorrência do ácido aracdônico, que até o presente momento não havia sido relatada na espécie em estudo.

Bixa orellana is a native plant of Brazil, popularly known as annatto. The purpose of this study was to isolate the oil from the seeds of B. orellana and from it, identify the antioxidant tocotrienol and quantify its fatty acids. The extraction of the oil was performed in a Soxhlet apparatus using hexane as solvent. The tocotrienol was isolated by chromatographic methods and identified by spectrometric methods NMR 1H and 13C. The fatty acids present in the oil were quantified by gas chromatography coupled with mass spectrometry. Results demonstrated the presence of δ-tocotrienol and among the fatty acids, the arachidonic acid was present, a substance which so far had not been previously reported.

Antioxidant enzyme activity and malondialdehyde levels can be modulated by Piper betle, tocotrienol rich fraction and Chlorella vulgaris in aging C57BL/6 mice

OBJECTIVE: The aim of this study was to determine the erythrocyte antioxidant enzyme activity and the superoxide dismutase, catalase, glutathione peroxidase, and plasma malondialdehyde levels in aging mice and to evaluate how these measures are modulated by potential antioxidants, including the tocotrienol-rich fraction, Piper betle, and Chlorella vulgaris. METHOD: One hundred and twenty male C57BL/6 inbred mice were divided into three age groups: young (6 months old), middle-aged (12 months old), and old (18 months old). Each age group consisted of two control groups (distilled water and olive oil) and three treatment groups: Piper betle (50 mg/kg body weight), tocotrienol-rich fraction (30 mg/kg), and Chlorella vulgaris (50 mg/kg). The duration of treatment for all three age groups was two months. Blood was withdrawn from the orbital sinus to determine the antioxidant enzyme activity and the malondialdehyde level. RESULTS: Piper betle increased the activities of catalase, glutathione peroxidase, and superoxide dismutase in the young, middle, and old age groups, respectively, when compared to control. The tocotrienol-rich fraction decreased the superoxide dismutase activity in the middle and the old age groups but had no effect on catalase or glutathione peroxidase activity for all age groups. Chlorella vulgaris had no effect on superoxide dismutase activity for all age groups but increased glutathione peroxidase and decreased catalase activity in the middle and the young age groups, respectively. Chlorella vulgaris reduced lipid peroxidation (malondialdehyde levels) in all age groups, but no significant changes were observed with the tocotrienol-rich fraction and the Piper betle treatments. CONCLUSION: We found equivocal age-related changes in erythrocyte antioxidant enzyme activity when mice were treated with Piper betle, the tocotrienol-rich fraction, and Chlorella vulgaris. However, Piper betle treatment showed increased antioxidant enzymes activity during aging.

The Effect of Curcumin and Tocotrienol on the Development of Eye Disease.

There is growing evidence that inflammation may be one of the causative factors of many chronic diseases especially which is related to eyes such as cataract,age-related macular degeneration and uveitis. Several cytokines such as IL-1,IL-6, RANKL, OPG, and M-CSF were implicated in the pathogenesis of chronic diseases. Anticytokine therapy using cytokine antagonists such as IL-receptor antagonist and TNF-binding protein was able to suppress the activity of the respective cytokines and prevent bone loss. Few animal studies have shown that vitamin E in the forms of palm-derived tocotrienol and α-tocopherol may prevent chronic eye disease in rat models by suppressing IL-1 and IL-6. Free radicals are known to activate transcription factor NFκB which leads to the production of bone resorbing cytokines. Tocotrienol, a potent antioxidant, may be able to neutralize free radicals before they could activate NFκB, therefore suppressing cytokine production and inflammatory reaction. Curcumin is widely reported to have potent anti-oxidative, anti-inflammatory and anticarcinogenic effects. The anti-inflammatory action of curcumin seems to be closely related to inhibition of TNF-α and other inflammatory cytokines production and suppression of NF-κB activation by blocking phosphorylation of inhibitory factor I-kappa B kinase (IκB) Tocotrienol and curcuminhave also been shown to inhibit COX-2, the enzyme involved in inflammatory reactions of the these studied, tocotrienol seemed to be better than tocopherols in terms of its ability to suppress inflammation induced by cytokines.