Simvastatin and vitamin E effects on cardiac and hepatic oxidative stress in rats fed on high fat diet

High fat diet (HFD) is a common cause of metabolic syndrome and type 2 diabetes mellitus. Published data showed that HFD and subsequent dyslipidemia are major triggers for oxidative stress. Forty-eight male Sprague–Dawley rats, weighing 170–200 g, were divided into six groups: control, control with vitamin E (100 mg/kg/day, i.p.), control with simvastatin (SIM) (10 mg/kg of body weight/day), HFD, HFD with vitamin E, and HFD with SIM. Standard and high cholesterol diets were given for 15 weeks and SIM and vitamin E were added in the last 4 weeks. In all rats, serum vitamin E, total cholesterol (TC), triglycerides (TG), low (LDL) and high (HDL) density lipoproteins, alanine (ALT) and aspartate (AST) transaminases, alkaline phosphatase (ALP), and gamma glutamyl transpeptidase (GGT) as well as cardiac and hepatic thiobarbituric acid-reactive substances (TBARS) and antioxidants (reduced glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT)) were measured. Also, electrocardiogram (ECG) was recorded. HFD significantly increased QTc interval, heart rate (HR), serum TC, TG, LDL, ALT, AST, ALP, GGT, liver TG, and cardiac and hepatic TBARS but decreased antioxidants and HDL, while SIM decreased HR, liver TG, serum TC, TG, and LDL and increased HDL in HFD rats. Vitamin E had no effect. Moreover, SIM and vitamin E decreased QTc interval, serum ALT, AST, ALP, GGT, and cardiac and hepatic TBARS and increased antioxidants in HFD rats. Histopathological observations confirm the biochemical parameters. SIM and vitamin E slow progression of hypercholesterolemia-induced oxidative stress in liver and heart and improve their functions.     

The influence of protonophore high concentration on the structure and functions of wheat root cells

Protonophore induced structural and functional changes in cells of excised roots of wheat seedlings have been investigated. The vector transfer of H+ inside the cells was accompanied by a decrease in energy supply of these cells (suppression of oxygen consumption and heat release), an output of K+ ions to the incubation medium, and by an increase in its pH value. The initial increase in heat release by roots (1 h) apparently reflects the process of dissipation of deltamicro H+ in plasma membrane. Within the first 5-10 min of exposure of 50 microM CCCP, changes in cell ultrastructure were observed that involved activation of Golgi apparatus, secretion of vesicle contents to the vacuole, and swelling of endoplasmic reticulum canals. Following a 2 h treatment with CCCP, structural and functional changes acquired a destructive character, and after 5-6 h of treatment with protonophore a complete desintegration of cell structure occurred demonstrating formations of myelin-like bodies, fragmentation of plasma membrane, and destruction of the nucleus. Thus, the protonophore induced proton excessive transport inside cells is fast and may cause an irreversible cell de-energization followed by serious disruption of ultrastructural organization of cells leading eventually to their death.     

维生素E对免疫功能的影响

适当剂量的维生素Ｅ（ＶＥ）,能增强抗体和补体的产生以及抗体对抗原的应答反应,促进淋巴细胞的增殖、分化和细胞因子的产生,提高免疫细胞的细胞毒作用和吞噬细胞的吞噬作用。ＶＥ缺乏或过量,能抑制机体的免疫机能,降低对疾病的抵抗能力。     

Dual effect of glucose on LDL oxidation: dependence on vitamin E

The aim of the present study was to determine the direct effect of glucose on LDL oxidation, a key step in the development of atherosclerosis. Purified human LDL were incubated with glucose (500 mg/dl) and LDL oxidation was started by adding CuCl(2) to the media. Glucose delayed the vitamin E consumption, but accelerated the formation of conjugated dienes and increased both the formation of thiobarbituric acid reacting substances (TBARS) and LDL electrophoretic mobility. When LDL were incubated with increasing concentrations of glucose and submitted to oxidation, the formation of conjugated dienes, TBARS, and the electrophoretic mobility increased in a concentration-dependent manner. When LDL was enriched with vitamin E, it showed a delay in the formation of conjugated dienes, even in the presence of glucose. To determine whether glucose had any effect on LDL oxidation, once the process was started and vitamin E consumed, LDL were submitted to oxidation and, at different times thereafter, glucose was added into the media. Under these conditions glucose also accelerated the LDL oxidation. In summary, present results show that in LDL submitted to oxidation, glucose delays the early phases of the oxidation, slowing the vitamin E consumption, but it accelerates the rate of LDL oxidation once LDL vitamin E has been consumed; the effect being concentration-dependent.     

高糖对线粒体乙醛脱氢酶2在大鼠心肌成纤维细胞中表达的影响

目的：观察心肌成纤维细胞是否存在线粒体乙醛脱氢酶2（ALDH2）的表达,探讨ALDH2在高糖诱导的心肌成纤维细胞引起纤维化发生中的作用。方法：原代培养心肌成纤维细胞,分为正常对照组（5.5 mmol/L）、正常+ALDH2激动剂Alda-1（20μmol/L）组、高糖组（30 mmol/L）、高糖+ Alda-1组。免疫荧光鉴定心肌成纤维细胞。各组细胞分别培养48 h后应用MTT法检测成纤维细胞增殖活力,RT-PCR和Western blot检测ALDH2 mRNA及蛋白的表达。结果：RT-PCR和Western blot结果显示心肌成纤维细胞ALDH2 mRNA和蛋白均有表达。与正常对照组相比,高糖组心肌成纤维细胞增殖能力提高（P < 0.01）,ALDH2蛋白表达下降（P < 0.05）;与高糖组相比,高糖+ Alda-1组心肌成纤维细胞增殖能力降低（P < 0.01）,ALDH2的蛋白表达增加（P < 0.05）。结论：心肌成纤维细胞存在ALDH2的表达,ALDH2激动剂Alda-1提高ALDH2的表达后可以抑制高糖引起的心肌成纤维细胞的增殖。     

Cerebral Na concentration, Na appetite and thirst of sheep: influence of somatostatin and losartan

Na and water intakes of Na-depleted sheep are influenced by changes in cerebral Na concentration. The effect of intracerebroventricular infusion of somatostatin or losartan, the ANG II type 1 receptor antagonist, on the Na appetite and thirst of Na-depleted sheep during infusions that decrease (intracerebroventricular hypertonic mannitol) or increase (intracerebroventricular or systemic hypertonic NaCl) cerebral Na concentration was investigated. Na intake was increased but water intake was unchanged during intracerebroventricular infusion of hypertonic mannitol. The increased Na appetite caused by intracerebroventricular infusion of hypertonic mannitol was decreased by concurrent intracerebroventricular infusion of either somatostatin or losartan, with somatostatin being most effective. Water intake was increased during intracerebroventricular infusion of hypertonic mannitol and somatostatin. Na intake was decreased and water intake was increased during systemic or intracerebroventricular infusion of hypertonic NaCl. Intracerebroventricular infusion of losartan blocked both (Na and water intake), whereas somatostatin did not influence either of these changes in intake. The results further consolidate a role for somatostatin and ANG II in the central mechanisms controlling Na appetite and thirst of sheep.     

Biological functions and metabolic fate of vitamin E revisited

Information accumulated lately has confirmed the essentiality of vitamin E for humans and provided a better understanding of its biological function and metabolic fate. The discovery of -tocopherol transfer protein, which preferentially binds to RRR--tocopherol, not only provides conclusive evidence of the essentiality of vitamin E for humans, but also sheds light on the superiority of RRR--tocopherol biologically over other isomers. The presence of tocopherol regeneration systems and multiple interdependent antioxidant systems is largely responsible for the lack of a widespread deficiency in humans and the difficulty to deplete vitamin E in the adult. The bulk of excess tocopherols consumed is excreted to feces unchanged or to urine with the side chain shortened but the chroman ring intact. The ability of dietary vitamin E to mediate mitochondrial superoxide generation affords a possible mode of action of vitamin E at the tissue levels. By decreasing the generation and/or the levels of reactive oxygen/nitrogen species, dietary vitamin E not only protects against oxidative damage, but also modulates the expression and/or activation of redox-sensitive biological response modifiers that regulate important cellular events.     

Influence of high vitamin E dosages on retinol and carotinoid concentration in body tissues and eggs of laying hens

The aim of the study was to contribute to the discussion of overdosing vitamin E in laying hens. A total of 45 laying hens, divided into 5 groups were fed diets supplemented with either 0; 100; 1000; 10 000 or 20 000 mg dl‐α‐tocopheryl acetate/kg diet over a period of 10 weeks. Concentrations of vitamins A and E were measured in plasma, various tissues and egg yolk. Furthermore egg yolk colour and some carotinoids were measured in egg yolks. None of the vitamin E doses significantly influenced performance of the hens. As expected, vitamin E concentration in plasma, all tissue samples and egg yolk was significantly increased with increasing tocopherol content in the diet. The egg yolk showed the highest vitamin E concentration, followed by liver and muscles. Feeding 1000 mg α‐tocopheryl acetate per kg diet resulted in an increase of vitamin A concentration in the liver. Very high doses (10 000 and 20 000 mg/kg diet) significantly decreased retinol concentration in the liver and egg yolk, as well as carotinoid concentration in the egg yolk. The lower carotinoid concentration in egg yolk resulted in a decreased intensity of egg yolk colour. A prooxidative and/or competitive effect of very high doses of vitamin E with other fat soluble substances has been discussed.     

Profibrotic effects of endothelin-1 via the ETA receptor in cultured human cardiac fibroblasts.

BACKGROUND/AIMS: Endothelin-1 (ET-1) has been implicated in pathologic remodelling and tissue repair processes in the heart. We investigated the effects of ET-1 on growth and collagen synthesis responses in cardiac fibroblasts isolated from human hearts. We also studied the receptor subtype(s) mediating such responses and the factors regulating their expression. METHODS: Fibroblasts were isolated from cardiac transplant recipient hearts and characterised by immunocytochemistry. Serum-starved cells were exposed to ET-1 and incorporation of [3H]proline and thymidine were measured as indexes of collagen and DNA synthesis respectively. Blocking experiments utilised the selective ETA receptor antagonist BQ123 and the ETB antagonist BQ788. RESULTS: ET-1 elicited a potent collagen synthesis response in cardiac fibroblasts, with a maximum 29+/-5% increase that was abolished by BQ123. Cardiac fibroblasts responded to ET-1 with a concentration-dependent decrease in DNA synthesis rate. The effects of ET-1 were similar to those of TGF-beta. Radioligand binding studies revealed the presence of high-affinity ET-1 binding sites on these cells, which were upregulated by treatment with the growth factors PDGF and EGF but downregulated by TGF-beta. CONCLUSIONS: These results therefore implicate ET-1 as a trophic agent in the human heart with the ability to influence the development of cardiac fibrosis.     

Rumen-protected choline and vitamin E supplementation in periparturient dairy goats: effects on milk production and folate, vitamin B12 and vitamin E status

We investigated the effects of rumen-protected choline (RPC) and vitamin E (VITE) administration on milk production and status of folate, vitamin B12 and vitamin E during the periparturient period of dairy goats. Forty-eight Saanen multiparous goats were selected for the 72-day experiment, being moved to a maternity pen 30 days before expected parturition and assigned to one of the four experimental groups: control (CTR), no choline or vitamin E supplementation; choline (RPC), supplemented with 4 g/day choline chloride in rumen-protected form; vitamin E (VITE), supplemented with 200 IU/day vitamin E in rumen-protected form; and choline and vitamin E (RPCE), supplemented with 4 g/day RPC chloride and 200 IU/day vitamin E. Supplements were administered individually before the morning feed to ensure complete consumption, starting 30 days before kidding and continuing for 35 days after. During the experiment, milk yield and 4% fat-corrected milk (FCM) yield were, respectively, 210 and 350 g/day higher in RPC-supplemented goats than in non-supplemented goats. Milk fat concentration and fat yield were also increased by RPC treatment. Milk yield and composition were unaffected by vitamin E supplementation. There were no significant interactions between RPC and VITE for any of the variables measured. Plasma metabolites did not differ between treatments before and after kidding except that plasma folate at parturition was higher in RPC-supplemented goats. Neither choline nor vitamin E affected vitamin B12 plasma concentrations, while a time effect was evident after the second week of lactation, when B12 levels in each treatment group started to increase. Vitamin E administration resulted in plasma α-tocopherol levels that were 2 to 2.5 times higher than in non-supplemented goats. Overall, these results suggest that greater choline availability can improve milk production and methyl group metabolism in transition dairy goats.     

Non-antioxidant molecular functions of alpha-tocopherol (vitamin E)

alpha-Tocopherol (the major vitamin E component) regulates key cellular events by mechanisms unrelated with its antioxidant function. Inhibition of protein kinase C (PKC) activity and vascular smooth muscle cell growth by alpha-tocopherol was first described by our group. Later, alpha-tocopherol was shown to inhibit PKC in various cell types with consequent inhibition of aggregation in platelets, of nitric oxide production in endothelial cells and of superoxide production in neutrophils and macrophages. alpha-Tocopherol diminishes adhesion molecule, collagenase and scavenger receptor (SR-A and CD36) expression and increases connective tissue growth factor expression.     

Influence of vitamin E supplementation and basal diet on the vitamin E status, performance and tissue fatty acid concentration in lambs

In order to determine the effect of dietary vitamin E level and basal diet on vitamin E status, performance and tissue fatty acid content, five groups of eight Suffolk × Charollais wether lambs with an initial live weight of 28.4 (s.d. 1.6) kg were allocated to one of five concentrate-based diets supplemented with all-rac-α-tocopheryl acetate to contain 30 mg (C-30), 60 mg (C-60), 120 mg (C-120), 250 mg (C-250) or 500 mg (C-500) α-tocopheryl acetate/kg dry matter (DM), for 63 days. Two additional groups of eight lambs entered the study at 31.2 (s.d. 3.3) kg and were fed grass silage and 400 g/day concentrate for 56 days, with the whole diet providing the equivalent of 60 mg (S-60) or 500 mg (S-500) α-tocopheryl acetate/kg DM. Lambs were weighed and blood samples obtained by venipuncture weekly. Dietary vitamin E level did not affect performance (P > 0.05), but lambs fed grass silage grew more slowly (P < 0.001) and had a higher (P < 0.001) feed conversion ratio (kg feed/kg gain) than those fed concentrates. At day 0 plasma α-tocopherol concentrations were 0.8 μg/ml and did not differ between treatments (P > 0.05). Plasma α-tocopherol concentrations then decreased in all lambs except for those fed S-500, which increased, and at slaughter were (μg/ml) 0.07, 0.23, 0.39, 0.76 and 1.57 in C-30, C-60, C-120, C-250 and C-500 and 1.18 and 1.93 in S-60 and S-500, respectively. At slaughter, muscle and liver α-tocopherol concentrations were in the deficiency range for lambs fed C-30, C-60 or C-120, whereas plasma creatine kinase and tissue polyunsaturated fatty acids were unaffected by dietary vitamin E level, but creatine kinase levels were higher (P < 0.05) and glutathione peroxidise levels lower (P < 0.001) in lambs fed grass silage than concentrates alone. Muscle and liver α-tocopherol concentrations were 1.8- and 4.1-fold higher in lambs fed S-60 than C-60, but there was less of a difference between lambs fed S-500 or C-500 with muscle and liver differences of 0.4- and 0.7-fold, respectively. Tissue n-3 polyunsaturated fatty acid concentrations were higher (P < 0.05) and n-6 fatty acids lower in lambs receiving the grass silage compared to concentrate-based diets, but were not affected by dietary vitamin E level. It is concluded that lower plasma and tissue levels of α-tocopherol are present in lambs supplemented with all-rac-α-tocopheryl acetate on a concentrate compared to a mixed diet of silage and concentrates, and that normal growth can be achieved at tissue levels previously considered to represent deficiency.     

Sheep deficient in vitamin E preferentially select for a feed with a higher concentration of vitamin E

Given the capacity of ruminants to modify diet selection based on metabolic needs, we hypothesised that, when given a choice, lambs experiencing a vitamin E deficiency would consume more of a vitamin E-enriched feed than lambs not deficient in vitamin E. Fifty-six Dohne Merino lambs were divided into two groups and fed either a vitamin E-deficient diet over 40 days to induce low plasma vitamin E or a vitamin E-enriched diet to induce high plasma vitamin E. The lambs were then offered a choice of vitamin E-enriched and vitamin E-deficient pellets. For half of the animals, the enriched diet was paired with strawberry flavour and the deficient diet was paired with orange flavour, while the reverse pairings were offered to the others. Lamb preference for the diets was measured daily for the following 15 days. There was a three-way interaction between the high and low vitamin E treatment groups×vitamin E content and type of flavour in the feed×time (days). The lambs preferred pellets flavoured with strawberry but this preference changed to orange flavour in vitamin E-deficient lambs if the orange flavour was paired with high vitamin E. Lambs without a deficiency continued to prefer strawberry-flavoured pellets, regardless of the vitamin E concentrations in the pellets. It is possible that self-learning contributed to the low vitamin E group of lambs changing preference to orange flavour in order to consume more vitamin E, presumably to remediate the deficiency.     

Comparative metabolic effects of fructose and glucose in human fibroblast cultures

Summary The comparative metabolic effects of fructose and glucose were determined in human fibroblast cultures. Cells were grown in four different media containing 5.5 and 27.5 mM of glucose and fructose, respectively. For these two hexoses, we compared their uptake, consumption, and conversion into¹⁴CO₂ and¹⁴C-lipids. D-Fructose was taken up in fibroblasts by an unsaturable process and its consumption was much smaller than that ofD-glucose. Whatever the experimental procedure, the glycogen content of cells grown in fructose media was significantly lower than of those grown in glucose media. Labeling of fructose and glucose with¹⁴C showed that more carbon from fructose than from glucose was incorporated into CO₂ and glycerolipids. The relative distribution of¹⁴C in the different lipid fractions was similar for both hexoses. These results indicated that the pathways of intermediary metabolism in fibroblast cultures were influenced by the nature of the carbohydrate present in the culture medium and that fructose was a better lipogenic substrate than glucose in human fibroblast cultures. This work was supported by grants for the Institut National de la Santé et de la Recherche Medicale (ATP 82-79-114).     

维生素E对暴露于高温及PM2.5中COPD大鼠呼吸系统的保护性作用*

目的:探讨维生素E改善暴露于高温与PM_2.5对慢性阻塞性肺疾病(COPD)大鼠呼吸系统功能损伤的作用。方法:将54只7周龄SPF级雄性Wistar大鼠随机分成9个实验组(n=6),利用烟草烟雾和气管内滴注脂多糖建立COPD大鼠模型,而后对其进行PM_2.5(0 mg/ml、3.2 mg/ml)气管滴注染毒和维生素E腹腔注射(20 mg/ml)干预,随后高温组进行高温(40℃)暴露,每天一次(8 h),持续3 d。末次暴露后检测肺功能,HE染色制作肺组织病理切片,采用试剂盒测定大鼠肺组织中诱导性一氧化氮合酶(iNOS)、肿瘤坏死因子(TNF-α)、单核细胞趋化蛋白1(MCP-1)的表达水平。结果:与对照组相比,高温和PM_2.5暴露使COPD大鼠肺功能降低(P＜0.05),各PM_2.5染毒组中MCP-1含量均有显著升高(P＜0.05),高温组中iNOS活性显著增高(P＜0.05);与单纯PM_2.5染毒组相比,在常温健康和高温COPD维生素E干预组的TNF-α含量均显著降低(P＜0.05),三组维生素干预组中MCP-1含量均显著降低(P＜0.05),高温COPD维生素干预组中iNOS活性显著降低(P＜0.05)。结论:高温与PM_2.5可引起并进一步加重COPD大鼠的炎症反应,维生素E作为一种抗氧化剂,可明显改善上述损伤作用,从而保护机体。     

Amelioration of glucose induced hemolysis of human erythrocytes by vitamin E

Cells under aerobic condition are always threatened with the insult of reactive oxygen species, which are efficiently taken care of by the highly powerful antioxidant systems of the cell. The erythrocytes (RBCs) are constantly exposed to oxygen and oxidative stress but their metabolic activity is capable of reversing the injury under normal conditions. In vitro hemolysis of RBCs induced by 5, 10 and 20 mM glucose was used as a model to study the free radical induced damage of biological membranes in hyperglycemic conditions and the protection rendered by vitamin E on the same. RBCs are susceptible to oxidative damage, peroxidation of the membrane lipids, release of hemoglobin (hemolysis) and alteration in activity of antioxidant enzymes catalase and superoxide dismutase. The glucose induced oxidative stress and the protective effect of vitamin E on cellular membrane of human RBCs manifested as inhibition of membrane peroxidation and protein oxidation and restoration of activities of superoxide dismutase and catalase, was investigated.Thiobarbituric acid reactive substances are generated from decomposition of lipid peroxides and their determination gives a reliable estimate of the amount of lipid peroxides present in the membrane. Vitamin E at 18 μg/ml (normal serum level) strongly enhanced the RBC resistance to oxidative lysis leading to only 50–55% hemolysis in 24 h, whereas RBCs treated with 10 and 20 mM glucose without vitamin E leads to 70–80% hemolysis in 24 h. Levels of enzymic antioxidants catalase, superoxide dismutase and nonenzymic antioxidants glutathione showed restoration to normal levels in presence of vitamin E. The study shows that vitamin E can protect the erythrocyte membrane exposed to hyperglycemic conditions and so a superior antioxidant status of a diabetic patient may be helpful in retarding the progressive tissue damage seen in chronic diabetic patients.