Alpha-tocopherol protects against oxidative damage to lipids of the rod outer segments of the equine retina

Oxidative stress is a possible risk factor for eye diseases. Lipid peroxidation is one of the major events induced by oxidative stress and is particularly active in polyunsaturated fatty acid (PUFA)-rich biomembranes. This work evaluated endogenous lipid antioxidants, in vitro non-enzymatic lipid peroxidation of rod outer segment membranes (ROS), the fatty acid composition during oxidative damage of total lipids from equine retina and ROS, and the protective action of alpha-tocopherol (alpha-Toc). The major lipid soluble antioxidant was alpha-Toc followed by retinoids and carotenoids. The retina contained a high percentage of PUFAs, mainly docosahexaenoic acid (22:6n-3) and arachidonic acid (20:4n-6). Lipid peroxidation of the equine ROS, induced by Fe(2+)-ascorbate, was monitored using chemiluminescence (CL) with or without pre-treatment with alpha-Toc. With alpha-Toc pre-treatment, CL values were significantly decreased. The most abundant fatty acid was 22:6n-3. After 3h incubation, 95% of total PUFAs were destroyed by peroxidation, whereas in alpha-Toc pre-treated ROS the percentage was significantly decreased. The results show that the retina has an endogenous lipid soluble antioxidant system. ROS were highly sensitive to oxidative damage, since their fatty acid composition was markedly modified during the lipid peroxidation process. The protective role of alpha-Toc as an antioxidant was evident and it could be used in the treatment of equine ocular diseases in which free radicals are involved.     

The effect of different concentrations of linseed oil or fish oil in the maternal diet on the fatty acid composition and oxidative status of sows and piglets

N‐3 polyunsaturated fatty acids (PUFA) are essential for foetal development. Hence, including n‐3 PUFA in the sow diet can be beneficial for reproduction. Both the amount and form (precursor fatty acids vs. long chain PUFA) of supplementation are important in this respect. Furthermore, including n‐3 PUFA in the diet can have negative effects, such as decreased arachidonic acid (ARA) concentration and increased oxidative stress. This study aimed to compare the efficacy to increase eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) concentrations in the piglet, when different concentrations of linseed oil (LO, source of precursor α‐linolenic acid) or fish oil (FO, source of EPA and DHA) were included in the maternal diet. Sows were fed a palm oil diet or a diet including 0.5% or 2% LO or FO from day 45 of gestation until weaning. Linoleic acid (LA) was kept constant in the diets to prevent a decrease in ARA, and all diets were supplemented with α‐tocopherol acetate (150 mg/kg) and organic selenium (0.4 mg/kg) to prevent oxidative stress. Feeding 0.5% LO or 0.5% FO to the sows resulted in comparable EPA concentrations in the 5‐day old piglet liver, but both diets resulted in lower EPA concentrations than when 2% LO was fed. The highest EPA concentration was obtained when 2% FO was fed. The DHA level in the piglet liver could only be increased when FO, but not LO, was fed to the sows. The 2% FO diet had no advantage over the 0.5% FO diet to increase DHA in the piglet. Despite the constant LA concentration in the sow diet, a decrease in ARA could not be avoided when LO or FO were included in the diet. Feeding 2% FO to the sows increased the malondialdehyde concentration (marker for lipid peroxidation) in sow plasma, but not in piglets.     

The influence of sex and gonadectomy on hepatic and brain fatty acid composition,lipogenesis and β‐oxidation

The aim of this study was to investigate the influence of sex and castration of rats on liver and brain fatty acid profile and liver mRNA expression of genes involved in lipogenesis and β‐oxidation. Castration significantly increased body weight and liver index and decreased serum triglyceride content in the female rats. The fatty acid composition of the liver tissue was influenced by sex and castration. Male rats had higher content of C16:0, C18:1n7, C18:2n6 and C22:5n3, while female rats had higher content of C18:0, C20:4n6 and C22:6n3. Castration of male rats decreased differences caused by sex for C18:2n6, C20:4n6 and C22:6n3. Values for C16:1n7 were higher in the castrated male rats in comparison with all other groups. Liver phospholipids showed a distribution of fatty acids similar to the total lipids. Brain total lipids and phospholipids were not influenced by sex or castration. Castration increased ?6D gene expression in both the sexes, while ?5D and ?9D increased in females and males respectively. Gonadectomy increased expression of the FASN gene in the females and decreased CPT1 and ACOX1 gene expression in the liver tissue of male rats. The observed results of lipid peroxidation, measured by TBARS, were the lowest in the intact females in comparison with all other groups. In conclusion, sex strongly influences both SFA and PUFA in liver tissue, and castration decreases these differences only for PUFA. Castration also influences the expression of the genes involved in lipid metabolism differently in male and female rats, with an increase in lipogenic genes in female rats and a decrease in key genes for mitochondrial and peroxisomal β‐oxidation in male rats.     

猪甘油三酯水解酶和激素敏感脂酶基因表达规律的研究

利用半定量RT-PCR法分析比较了甘油三酯水解酶(Triacylglycerol hydrolase,TGH)和激素敏感脂酶(Hormone-sensitive lipase,HSL)基因在不同猪种、不同发育阶段及不同部位脂肪组织中转录表达的差异,探讨其在猪脂肪组织的表达规律。结果显示,脂肪型个体TGHmRNA表达丰度显著低于瘦肉型和杂交型个体,成年猪较初生仔猪低,皮下、腹膜和内脏脂肪组织中TGH表达量依次递增;其变化规律与HSL相同。此外,对分离培养的原代前体脂肪细胞通过诱导分化和油红O染色区分分化状态,分析TGHmRNA表达的时序变化,发现TGH在前脂肪细胞中不转录表达,诱导分化后开始表达,且在诱导分化第4天表达量最高,分化第10天表达量下降,达到峰值的时间较HSL早。结果表明,TGH的表达与个体肥胖程度、年龄、脂肪组织部位以及脂肪细胞分化程度相关,同时,在脂肪细胞分化过程中,TGH表达峰值早于HSL,提示TGH在脂肪细胞发育过程中可能较早承担基础脂解作用。     

长链n-3多不饱和脂肪酸对肠上皮细胞促炎细胞因子基因mRNA表达的影响

本试验旨在探讨长链n-3多不饱和脂肪酸(LC n-3 PUFA)对肠上皮细胞促炎细胞因子基因mRNA表达的影响.试验选用大鼠肠上皮细胞系IEC-6细胞为模型,分为4个处理,分别为对照、脂多糖(LPS,1μg/mL)、LPS(1μg/mL)+二十二碳六烯酸(DHA,100 μmol/L)和LPS(l μg/mL)+二十碳五烯酸(EPA,100μmol/L),每个处理3个重复,每孔为1个重复.细胞先用DHA、EPA或等量二甲基亚砜(DHA和EPA的溶剂,对照)预处理48h,再用LPS处理3h,收集细胞提取总RNA,采用实时定量PCR方法分析肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)和白介素-6(IL-6)的基因mRNA表达水平的差异.结果表明:LPS极显著上调了细胞中TNF-α、IL-1 β和IL-6的基因mRNA表达水平(P＜0.01),EPA均极显著或显著削弱了细胞内LPS诱导的TNF-α(P＜0.01)、IL-1β(P ＜0.01)和IL-6的基因mRNA水平(P＜0.05)的上调,而DHA仅显著削弱了细胞内LPS诱导的IL-1β的基因mRNA水平的上调(P＜0.05).结果提示,LC n-3 PUFA在肠上皮细胞中具有抗炎作用,且在本试验条件下EPA的抗炎效果要优于DHA.     

Poultry fat decreased fatty acid transporter protein mRNA expression and affected fatty acid composition in chickens

Background

A study was undertaken to examine the effects of poultry fat (PF) compared with those of soybean oil (SBO) on intestinal development, fatty acid transporter protein (FATP) mRNA expression, and fatty acid composition in broiler chickens. A total of 144 day-old male commercial broilers were randomly allocated to 2 treatment groups (6 replicates of 12 chicks for each treatment) and fed isocaloric diets containing 3.0% PF or 2.7% SBO at 0 to 3 wk and 3.8% PF or 3.5% SBO at 4 to 6 wk, respectively.

Results

PF had no influence on intestinal morphology, weight, or DNA, RNA, or protein concentrations at 2, 4, and 6 wk of age. However, compared with SBO, PF significantly decreased FATP mRNA abundance at 4 wk (P = 0.009) and 6 wk of age (P < 0.001); decreased liver fatty acid-binding protein (L-FABP) mRNA abundance at 6 wk of age (P = 0.039); and decreased C18:2 (P = 0.015), C18:3 (P < 0.001), C20:2 (P = 0.018), Σ-polyunsaturated fatty acids (Σ-PUFA) (P = 0.020), and the proportion of PUFA (P < 0.001) in the intestinal mucosa and decreased C18:2 (P = 0.010), C18:3 (P < 0.001), C20:2 (P < 0.001), Σ-PUFA (P = 0.005), and the proportion of PUFA (P < 0.001) in breast muscle at 6 wk of age.

Conclusions

PF decreases FATP and L-FABP mRNA expression and decreased the proportion of PUFA in the intestinal mucosa and breast muscle.     

日粮中添加整粒油籽对绵羊体脂脂肪酸组成的影响

日粮中分别添加相当于40 g粗脂肪的油菜籽(canola)、油葵籽、红花籽和亚麻籽,饲喂成年母羊35 d后屠宰,用气相色谱技术测定了背最长肌、尾部、背膘和肾脏4个部位脂肪的脂肪酸(FA)组成,结果表明:1)红花籽可显著降低肌肉中的短链、长链饱和脂肪酸及总饱和脂肪酸(SFA)的比例,而增加单不饱和脂肪酸(MUFA)和总不饱和脂肪酸(UFA)的比例,而添加油菜籽对肌肉的FA无显著影响;2)油葵显著降低了尾部短链和中链脂肪酸,提高了多不饱和脂肪酸(PUFA)与SFA的比值(P:S),油菜籽可显著降低尾部短链脂肪酸;3)添加亚麻籽有增加所有组织UFA和P:S值,降低SFA的趋势,但只有背膘脂肪中的PUFA显著增高;4)各类油籽对脂肪组织中的总SFA和MUFA无明显影响,其中肾脏脂肪受影响最小.     

Porcine leptin alters isolated adipocyte glucose and fatty acid metabolism

This study examined if leptin can acutely affect glucose or fatty acid metabolism in pig adipocytes and whether leptin's actions on lipogenesis are manifested through interaction with insulin or growth hormone. Subcutaneous adipose tissue was obtained from approximately 55 kg crossbred barrows at the USDA abattoir. Isolated adipocytes were prepared using a collagenase procedure. Experiments assessed U-14C-glucose or 1-14C-palmitate metabolism in isolated adipocytes exposed to: basal medium (control), 100 nM insulin, 100 ng/ml porcine growth hormone, 100 ng/ml recombinant porcine leptin, and combinations of these hormones. Treatments were performed in triplicate and the experiment was repeated with adipocytes isolated from five different animals. Cell aliquots (250 microl) were added to 1 ml of incubation medium, then incubated for 2h at 37 degrees C for measurement of glucose and palmitate oxidation or incorporation into lipid. Incubation of isolated adipocytes with insulin increased glucose oxidation rate by 18% (P<0.05), while neither growth hormone nor leptin affected glucose oxidation (P>0.5). Total lipid synthesis from glucose was increased by approximately 25% by 100 nM insulin or insulin+growth hormone (P<0.05). Insulin+leptin reduced the insulin response by 37% (P<0.05). The combination of all three hormones increased total lipid synthesis by 35%, relative to controls (P<0.05), a rate similar to insulin alone. Fatty acid synthesis was elevated by insulin (32%, P<0.05) or growth hormone (13%, P<0.05). Leptin had no effect on fatty acid synthesis (P>0.05). Leptin reduced the esterification rate by 10% (P<0.05). Growth hormone and insulin could overcome leptin's inhibition of palmitate esterification (P>0.05).     

Polyunsaturated fatty acids influence inflammatory markers in a cellular model for canine osteoarthritis

Although it is well recognized that dietary supplementation with fish oil improves clinical symptoms in dogs suffering from osteoarthritis, the molecular basis for the dietary benefit is not yet completely resolved in dogs. This study was designed to further clarify how polyunsaturated fatty acids (PUFA) affect key factors of cartilage degeneration in a canine cell culture system mimicking osteoarthritis. Canine chondrocytes were incubated either without or with 10 μm of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), arachidonic acid (AA) or 3.6 μm ibuprofen (Ibu) as positive control for 6 days. After the supplementation, cells were stimulated with 10 ng/ml interleukin‐1β (IL‐1β) for another 48 hr to induce osteoarthritic changes, or left unstimulated. We analysed fatty acid uptake via gas–liquid chromatography, nitric oxide (NO) production via Griess assay, prostaglandin E (PGE) production via ELISA and relative gene expression of several cartilage matrix proteinases, inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 via RT‐qPCR. After supplementation, the chondrocytes rapidly incorporated the PUFA into their fatty acid pools. The stimulation with IL‐1β caused a marked increase of most of the inflammatory markers measured. N‐3 PUFA EPA reduced IL‐induced gene expression of iNOS and corresponding production of NO. N‐6 PUFA AA also decreased iNOS and NO, but furthermore lowered gene expression of matrix metalloproteinase‐3. On the other hand, AA upregulated the aggrecanase ADAMTS‐5 and augmented the release of PGE. The effect of n‐3 PUFA DHA turned out to be negligible. Our results reveal molecular mechanisms by which PUFA affect degenerative joint disease in dogs. Of particular importance is that not only EPA but also AA decreased several inflammatory markers in our model. Thus, we conclude that an appropriate balance of both n‐3 and n‐6 fatty acids deserves more attention in dietary interventions.     

INS、GLN和LP对体外培养牛脂肪细胞内HSL、LP的mRNA丰度的影响

为了阐明胰岛素（INS）、胰高血糖素（GLN）和瘦蛋白（LP）对牛脂肪细胞内激素敏感酯酶（HSL）、LP的mRNA表达的调控作用，本试验采用体外原代培养牛脂肪细胞的培养液中添加不同浓度的胰岛素（0、5、10、20、50、100mmol／L）、胰高血糖素（O、25、100、200、500、1000pg／mL）和瘦蛋白（O、2．5、5、10、50、100ng／mL），培养12h后，应用建立的实时荧光定量PCR（FQ-PCR）方法检测牛脂肪细胞内HSL、LP的mRNA丰度。结果表明：胰岛素对牛脂肪细胞内HSL的mRNA表达呈现剂量依赖性抑制作用，对LP的mRNA表达呈现剂量依赖性促进作用。胰高血糖素对牛脂肪细胞内HSL的mRNA表达呈现剂量依赖性促进作用，对牛脂肪细胞内LP的mRNA表达无作用。瘦蛋白对牛脂肪细胞内HSL的mRNA表达先呈现剂量依赖性促进作用，后呈现抑制作用。但对牛脂肪细胞内LP的mRNA表达呈现剂量依赖性抑制作用。     

饲喂不同油脂对黄羽肉鸡肌肉组织中脂肪酸组成的影响

选用 8 3日龄的雌性广西黄羽肉鸡 90只 ,随机分成 5组 (每组设 2个重复 ,每个重复 9只 ,共 18只 ) ,研究饲粮中不同油脂对肉鸡肌肉组织脂肪酸组成的影响。 5组饲粮分别含 3%的富含饱和脂肪酸的棕榈油 (PO3组 ,对照组 )、 3% (LO3组 )和 5%的富含α 亚麻酸 (C18:3n 3,ALA)的亚麻油 (LO5组 )、 5%的富含长链n 3脂肪酸 (如C2 0 :5n 3,EPA ;C2 2 :6n 3,DHA)的鱼油 (FO5组 )、 5%的富含亚油酸(C18:2n 6 ,LA)和ALA的火麻仁油 (HO5组 )。饲喂 2 8d,取样测定胸肌和腿肌中的脂肪酸组成。结果表明 :PO3(对照组 )肌肉中的n 6 /n 3脂肪酸比值比例高于其他组 (P <0 0 5) ;与对照组相比 ,LO3组和LO5组以ALA和LA为主的多不饱和脂肪酸 (PUFA)增加 (P <0 0 5) ,ALA和LA在体内可转化为相应的长链脂肪酸 ;FO5组以长链n 3PUFA ,(如EPA、DPA、DHA)为主的PUFA含量增加 (P <0 0 5) ;HO5组以LA为主的n 6脂肪酸高于其他组 (P <0 0 5) ;长链PUFA (如DHA)易沉积于胸肌中 ,而LA ,ALA易沉积于腿肌中。由此可见 ,肉鸡肌肉组织脂肪酸的组成充分反映了饲粮油脂的脂肪酸组成 ,通过饲粮中添加不同脂肪酸组成的油脂可生产富含特定PUFA的鸡肉     

The influence of different doses of α-tocopherol and ascorbic acid on selected oxidative stress parameters in in vitro culture of leukocytes isolated from transported calves

The objective of the study was to assess in vitro the influence of various doses of two different antioxidants, α-tocopherol and ascorbic acid, on protective mechanisms against ROS in white blood cells obtained from calves exposed to transport, and to compare these results with those obtained from non-transported animals.The concentrations of lipid peroxidation products in leukocytes and in the retained medium were assessed by determining the level of ThioBarbituric Acid Reactive Substances (TBARS). Total antioxidant status in the leukocytes and the medium were estimated using a ferric-reducing ability of plasma (FRAP) assay. Leukocyte viability was determined using the trypan blue reduction test.The study demonstrated that after bovine leukocytes (WBC) were incubated in vitro with α-tocopherol and ascorbic acid, peroxidation intensity decreased and total antioxidant capacity increased. The results of the study reveal that these antioxidants in concentrations over 0.1 mg/ml have a major impact on free radical activity on bovine white blood cells and on cell viability during transport of animals.Based on this study, we suggest that incubation of the leukocytes with antioxidants decreases the oxidative stress development, which can be helpful in protection of the immunological cells during bovine respiratory disease.