高不饱和脂肪酸对草鱼稚鱼生长、脂质代谢的影响及其分子机理的研究
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摘要
为了深入探讨n-3 HUFA对草鱼稚鱼生长及脂代谢的影响,设计了含不同水平n-3 HUFA和不同比例DHA/EPA的半精制饲料对草鱼稚鱼进行饲喂,对试验鱼生长、生物学性状、体成分、肝胰脏抗氧化状态、脂代谢关键酶活力及转录水平等相关指标进行检测。结果表明:
(1)将200尾草鱼稚鱼(平均体质量(5.66±0.21)g)以20尾/缸的密度随机投放于10个养殖缸中,随机分为5组,进行为期75d的饲养试验,对照组草鱼稚鱼饲喂仅满足必需脂肪酸需求的饲料,试验组草鱼饲喂在满足必需脂肪酸需求的基础上,分别添加0.26%,0.52%,0.83%和1.13%的n-3 HUFA。饲养试验结束后,测量试验鱼体长和体质量,采取全鱼、肌肉、肝胰脏等进行相关指标测定和分析。在0-0.52%区间,随着n-3 HUFA水平升高,相对生长率,特定生长率,饲料效率和蛋白效率显著升高;过高水平,则出现负效应。腹腔脂肪组织和肝胰脏脂肪含量在0.52%组最低。组织脂肪酸组成反映了饲料脂肪酸组成。0.52%组肝胰脏LPL活力最高;在0-0.52%区域,MDH保持相对稳定;过高水平时,则显著下降。随着饲料n-3 HUFA水平升高(>0.52%),肝胰脏SOD活力和MDA含量显著升高,且呈现正相关。饲料n-3 HUFA水平为0.83%时,PPAR-α表达丰度最高。在0-0.83%区间,随着饲料n-3 HUFA水平升高,LPL表达丰度显著提高。
(2)将198尾草鱼稚鱼(平均体质量(17.6±1.2)g)以11尾/缸的密度随机投放于18个循环水养殖缸中,随机分为6组,进行为期38d的饲养试验,对照组草鱼稚鱼饲喂满足必需脂肪酸需求的饲料,试验组草鱼饲喂在满足必需脂肪酸需求的基础上,添加0.5% n-3 HUFA、DHA/EPA比率分别为4.93(试验Ⅰ组),2.05(试验Ⅱ组),1.08(试验Ⅲ组),0.49(试验IV组),0.21(试验V组)的饲料。饲养试验结束后,测量试验鱼体长和体质量,采取全鱼、肌肉、肝胰脏等进行相关指标测定和分析。试验V组草鱼稚鱼终末体质量、相对生长率和特定生长率显著高于对照组;试验I-V组的饲料效率和蛋白效率均显著高于对照组,处理组间无显著差异;试验Ⅱ和III组肌肉脂质含量显著高于对照组,试验I-V组肝胰脏脂质含量无显著差异,但均显著低于对照组;试验I-V组肝胰脏LPL活力显著对照组,处理组间无显著差异;G6PDH和MDH在试验组间差异不显著;对照组肝胰脏T-Aoc显著高于试验I、II、III和IV组,其余处理组差异不显著;试验组间肝胰脏CAT,SOD活性和MDA含量差异均不显著。
结论:草鱼饲料中添加适宜水平n-3 HUFA可促进生长,降低体脂沉积,调控体脂在组织间分配;本研究适宜水平为0.52%。n-3 HUFA可通过上调PPAR-α、LPL等脂代谢关键基因提高脂质利用能力。过高水平n-3 HUFA可导致草鱼稚鱼出现严重氧化应激,抑制生长。饲料DHA/EPA比率在4.93–0.21对草鱼稚鱼的生长、脂质沉积和抗氧化状态无显著性影响。
To evaluate the effect of n-3 HUFA on growth performance and lipid metabolism in grass carp. Two trial were designed, including dietary n-3 HUFA content (feed 75d) and DHA/EPA ratio (feed 38d). Some indicators were detected, including growth performance, fatty acid profiles, activities of lipogenesis-related enzymes, expression of lipid metabolism-related genes and anti-oxidation status. The result showed:
(1)a 75-day feeding experiment was conducted using five isonitrogenous and isoenergetic semi-purified diets containing 0% (control), 0.26%, 0.52%, 0.83% or 1.13% n-3 HUFA. Weight gain, specific growth rate, feed efficiency and protein efficiency increased by increasing the dietary n-3 HUFA content from 0% to 0.52% (P<0.05), and declined thereafter. Intraperitoneal fat content and the hepatopancreatic lipid levels were lowest in the 0.52% group (P<0.05). The tissue fatty acid level was well correlated with dietary n-3 HUFA content. Hepatic lipoprotein lipase (LPL) activity was significantly higher in the 0.52% group (P<0.05), while that of malate dehydrogenase (MDH) was stable in the 0–0.52% groups, and was significantly lower in the 1.13% group (P<0.05). Superoxide dismutase (SOD) activity increased significantly with increasing dietary HUFA content, consistent with the level of malondialdehyde (MDA) (P<0.05). Hepatic mRNA expression of peroxisome proliferator-activated receptor-α(PPAR-α) was greatest in the 0.83% group and that of the LPL gene increased with increasing dietary n-3 HUFA content up to 0.83% (P<0.05).
(2)A total of 198 grass carp juveniles (body weight (17.6±1.2)g) were randomly assigned into six groups with triplicate, total 18 tanks with 11 fish each. Grass carp juveniles were reared in glass-steel tanks (200-L volume per tank) with a continuous flow of water and aeration, temperature 28–30°C, dissolved O2 6–8mg/L and pH 7.5–7.7. Grass carp juveniles were fed for 38 days using six experimental diets (CK and I to V) with the same amount of DHA and EPA (0.5% of dried diet), but varying ratios of DHA to EPA (4.93,2.05,1.08,0.49 and 0.21 respectively) and a control diet (CK, no DHA and EPA included ). Final weight, weight gain and specific growth rate in treatment V were significantly higher in control group (P<0.05). The feed efficiency and protein efficiency were significantly higher in treatment I, II, III, IV and V (P <0.05), however, there were no statistically significant differences among treatment I to V. Muscle lipid content in treatment II and III were significant higher (P <0.05). Hepatopancreas lipid content in treatment I to V were significant lower than control group (P <0.05), however, there were no statistically significant differences in hepatopancreas lipid content among treatment I to V. The activities of lipoprotein lipase (LPL) were significantly higher in treatment I, II, III, IV and V (P <0.05), however, there were no statistically significant differences among treatment I to V. The activities of Glucose-6-phosphate dehydrogenase (G6PDH) and Malate Dehydrogenase (MDH) kept constant in all groups (P >0.05); The activities of total anti-oxidative capacity (T-AOC) were no statistically significant differences among groups. The activities of catalase (CAT), SOD and the content of MDA) kept constant in all groups (P >0.05). The activities of total antioxidative capacity (T-Aoc) in control group was significantly higher than treatment I to IV (P <0.05)
Conclude: adequate dietary n-3 HUFA supplementation significantly promotes growth performance and lipid metabolism through PPAR-αand LPL in freshwater fish grass carp. However, excess n-3 HUFA fortification may exert adverse effects, which might be due to oxidative stress. Adequate n-3 HUFA could adjust the distribution of body fat among tissues, also might induce oxidative stress, but the growth performance, lipid accumulation and antioxidation system not in?uenced by the ratios of DHA/EPA (4.93–0.21) .
(1)将200尾草鱼稚鱼(平均体质量(5.66±0.21)g)以20尾/缸的密度随机投放于10个养殖缸中,随机分为5组,进行为期75d的饲养试验,对照组草鱼稚鱼饲喂仅满足必需脂肪酸需求的饲料,试验组草鱼饲喂在满足必需脂肪酸需求的基础上,分别添加0.26%,0.52%,0.83%和1.13%的n-3 HUFA。饲养试验结束后,测量试验鱼体长和体质量,采取全鱼、肌肉、肝胰脏等进行相关指标测定和分析。在0-0.52%区间,随着n-3 HUFA水平升高,相对生长率,特定生长率,饲料效率和蛋白效率显著升高;过高水平,则出现负效应。腹腔脂肪组织和肝胰脏脂肪含量在0.52%组最低。组织脂肪酸组成反映了饲料脂肪酸组成。0.52%组肝胰脏LPL活力最高;在0-0.52%区域,MDH保持相对稳定;过高水平时,则显著下降。随着饲料n-3 HUFA水平升高(>0.52%),肝胰脏SOD活力和MDA含量显著升高,且呈现正相关。饲料n-3 HUFA水平为0.83%时,PPAR-α表达丰度最高。在0-0.83%区间,随着饲料n-3 HUFA水平升高,LPL表达丰度显著提高。
(2)将198尾草鱼稚鱼(平均体质量(17.6±1.2)g)以11尾/缸的密度随机投放于18个循环水养殖缸中,随机分为6组,进行为期38d的饲养试验,对照组草鱼稚鱼饲喂满足必需脂肪酸需求的饲料,试验组草鱼饲喂在满足必需脂肪酸需求的基础上,添加0.5% n-3 HUFA、DHA/EPA比率分别为4.93(试验Ⅰ组),2.05(试验Ⅱ组),1.08(试验Ⅲ组),0.49(试验IV组),0.21(试验V组)的饲料。饲养试验结束后,测量试验鱼体长和体质量,采取全鱼、肌肉、肝胰脏等进行相关指标测定和分析。试验V组草鱼稚鱼终末体质量、相对生长率和特定生长率显著高于对照组;试验I-V组的饲料效率和蛋白效率均显著高于对照组,处理组间无显著差异;试验Ⅱ和III组肌肉脂质含量显著高于对照组,试验I-V组肝胰脏脂质含量无显著差异,但均显著低于对照组;试验I-V组肝胰脏LPL活力显著对照组,处理组间无显著差异;G6PDH和MDH在试验组间差异不显著;对照组肝胰脏T-Aoc显著高于试验I、II、III和IV组,其余处理组差异不显著;试验组间肝胰脏CAT,SOD活性和MDA含量差异均不显著。
结论:草鱼饲料中添加适宜水平n-3 HUFA可促进生长,降低体脂沉积,调控体脂在组织间分配;本研究适宜水平为0.52%。n-3 HUFA可通过上调PPAR-α、LPL等脂代谢关键基因提高脂质利用能力。过高水平n-3 HUFA可导致草鱼稚鱼出现严重氧化应激,抑制生长。饲料DHA/EPA比率在4.93–0.21对草鱼稚鱼的生长、脂质沉积和抗氧化状态无显著性影响。
To evaluate the effect of n-3 HUFA on growth performance and lipid metabolism in grass carp. Two trial were designed, including dietary n-3 HUFA content (feed 75d) and DHA/EPA ratio (feed 38d). Some indicators were detected, including growth performance, fatty acid profiles, activities of lipogenesis-related enzymes, expression of lipid metabolism-related genes and anti-oxidation status. The result showed:
(1)a 75-day feeding experiment was conducted using five isonitrogenous and isoenergetic semi-purified diets containing 0% (control), 0.26%, 0.52%, 0.83% or 1.13% n-3 HUFA. Weight gain, specific growth rate, feed efficiency and protein efficiency increased by increasing the dietary n-3 HUFA content from 0% to 0.52% (P<0.05), and declined thereafter. Intraperitoneal fat content and the hepatopancreatic lipid levels were lowest in the 0.52% group (P<0.05). The tissue fatty acid level was well correlated with dietary n-3 HUFA content. Hepatic lipoprotein lipase (LPL) activity was significantly higher in the 0.52% group (P<0.05), while that of malate dehydrogenase (MDH) was stable in the 0–0.52% groups, and was significantly lower in the 1.13% group (P<0.05). Superoxide dismutase (SOD) activity increased significantly with increasing dietary HUFA content, consistent with the level of malondialdehyde (MDA) (P<0.05). Hepatic mRNA expression of peroxisome proliferator-activated receptor-α(PPAR-α) was greatest in the 0.83% group and that of the LPL gene increased with increasing dietary n-3 HUFA content up to 0.83% (P<0.05).
(2)A total of 198 grass carp juveniles (body weight (17.6±1.2)g) were randomly assigned into six groups with triplicate, total 18 tanks with 11 fish each. Grass carp juveniles were reared in glass-steel tanks (200-L volume per tank) with a continuous flow of water and aeration, temperature 28–30°C, dissolved O2 6–8mg/L and pH 7.5–7.7. Grass carp juveniles were fed for 38 days using six experimental diets (CK and I to V) with the same amount of DHA and EPA (0.5% of dried diet), but varying ratios of DHA to EPA (4.93,2.05,1.08,0.49 and 0.21 respectively) and a control diet (CK, no DHA and EPA included ). Final weight, weight gain and specific growth rate in treatment V were significantly higher in control group (P<0.05). The feed efficiency and protein efficiency were significantly higher in treatment I, II, III, IV and V (P <0.05), however, there were no statistically significant differences among treatment I to V. Muscle lipid content in treatment II and III were significant higher (P <0.05). Hepatopancreas lipid content in treatment I to V were significant lower than control group (P <0.05), however, there were no statistically significant differences in hepatopancreas lipid content among treatment I to V. The activities of lipoprotein lipase (LPL) were significantly higher in treatment I, II, III, IV and V (P <0.05), however, there were no statistically significant differences among treatment I to V. The activities of Glucose-6-phosphate dehydrogenase (G6PDH) and Malate Dehydrogenase (MDH) kept constant in all groups (P >0.05); The activities of total anti-oxidative capacity (T-AOC) were no statistically significant differences among groups. The activities of catalase (CAT), SOD and the content of MDA) kept constant in all groups (P >0.05). The activities of total antioxidative capacity (T-Aoc) in control group was significantly higher than treatment I to IV (P <0.05)
Conclude: adequate dietary n-3 HUFA supplementation significantly promotes growth performance and lipid metabolism through PPAR-αand LPL in freshwater fish grass carp. However, excess n-3 HUFA fortification may exert adverse effects, which might be due to oxidative stress. Adequate n-3 HUFA could adjust the distribution of body fat among tissues, also might induce oxidative stress, but the growth performance, lipid accumulation and antioxidation system not in?uenced by the ratios of DHA/EPA (4.93–0.21) .
引文
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