共轭亚油酸（CLA）对大黄鱼脂肪代谢、免疫、肉品质及PPAR基因表达的影响
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摘要
共轭亚油酸(CLA)是具有许多重要生理功能的天然不饱和脂肪酸之一。人体及动物实验表明,它具有降低脂肪、抗癌、抗动脉粥样硬化、增强肌体免疫能力、改善骨组织代谢等多种独特的生理功能。本研究在大黄鱼日粮中添加了不同水平的共轭亚油酸,探讨了共轭亚油酸对大黄鱼生长、脂肪代谢、免疫性能及对大黄鱼肌肉品质的影响,并对大黄鱼肝脏中的PPARa和PPARγ基因进行了克隆与表达,从分子水平上阐述了共轭亚油酸影响大黄鱼脂肪代谢的作用机制。主要研究内容和结果如下:
1.日粮CLA水平对大黄鱼生长性能和脂肪代谢的影响
本试验研究了日粮CLA对大黄鱼生长性能、体成分、脂质组成、脂肪代谢酶活性和脂肪酸成分的影响。在基础日粮中用CLA代替鱼油,添加水平分别为0%、1%、2%和4%(试验D1-D4组)。经过10周的饲喂,结果表明:饲喂1%的CLA能够提高大黄鱼的特定生长率(SGR)和肥满度(CF),饲喂高含量(2%和4%)的CLA则降低了SGR,但是四个试验组之间的差异不显著(P＞0.05)。日粮中添加CLA对大黄鱼的饵料系数(FCR)没有影响,但是肝体比(HSI)和脏体比(VSI)均有所提高,各处理组(D2-D4)之间差异也不显著(P＞0.05)。与对照组(D1)相比,大黄鱼鱼体的水分、粗蛋白和灰分均有所增加,粗脂肪含量有所下降,但是差异不显著(P＞0.05)。随着日粮中CLA水平的增加,大黄鱼肝脏和肌肉中的脂肪含量也有所降低(P＞0.05)。
与D1组相比,CLA能够显著降低血清和肝脏中甘油三酯(TG)的含量(P＜0.05)。血清中的TG含量随着CLA含量的增加显著降低,D2组与D3和D4组相比差异显著(P＜0.05)。在肝脏中,CLA处理组之间差异不显著(P＞0.05)。与此同时,CLA能够显著增加血清和肝脏中游离脂肪酸(FFA)的含量(P＜0.05)。在血清和肝脏中,D4组的FFA含量最高,并且与D1和D2组相比差异显著(P＜0.05)。随着日粮中CLA含量的增加,血清中胆固醇(CHO)和低密度脂蛋白胆固醇(LDL-C)含量也随着降低,CLA处理组与D1组相比有显著差异(P＜0.05)。血清中高密度脂蛋白胆固醇(HDL-C)的含量随着CLA含量的增加而增加。CLA处理组与D1组相比差异显著(P＜0.05),但是D2、D3和D4组之间无显著差异(P＞0.05)。
日粮中添加CLA能够显著降低大黄鱼血清和肝脏中肝脂酶(HL)和脂蛋白脂酶(LPL)的活性。在血清和肝脏中,D2、D3和D4组的HL和LPL活性与D1相比差异显著(P＜0.05),而且D2组与D3和D4相比也具有显著性差异(P＜0.05)。此外,CLA对大黄鱼血清和肝脏中葡萄糖-6-磷酸脱氢酶(G6PD)、脂肪合成酶(FAS)和苹果酸酶(ME)的活性有着不同程度的影响,1%CLA对血清和肝脏中的G6PD活性没有显著影响(P＞0.05)。但是高含量的CLA(2%和4%)能够显著降低血清和肝脏中的G6PD活性,并且与D1组相比差异显著(P＜0.05)。CLA能够显著降低大黄鱼血清中的脂肪酶(FAS)活性,与D1相比,D4组的FAS活性降低了16%,且差异显著(P＜0.05)。与此同时,CLA也能够降低大黄鱼肝脏中的FAS活性,但D1-D4组间差异不显著(P＞0.05)。大黄鱼血清和肝脏中苹果酸酶(ME)的活性随着日粮中CLA含量的增加而降低。在血清中,D3和D4组ME活性与D1相比差异显著(P＜0.05),而D2组与D1组相比差异不显著(P＞0.05)。在肝脏中,D2、D3和D4组之间的ME活性无显著差异(P＞0.05)。
CLA对肝脏和肌肉脂肪酸成分有显著的影响。饱和脂肪酸(SFA)的主要成分是C16:0;单不饱和脂肪酸(MUFA)的主要成分是C18:1 n-9。随着CLA含量的增加,肝脏和肌肉中SFA含量和多不饱和脂肪酸(PUFA)含量也随之增加,而MUFA含量随之降低,并且D1-D4之间差异显著(P＜0.05)。在PUFA中,EPA(C20:5 n-3)和DHA(C22:6 n-3)约占PUFA的65%和63%,并且随着CLA含量的增加而显著增加。此外,肝脏和肌肉中CLA的含量与日粮中CLA含量相同呈等比增加。
2.日粮CLA水平对大黄鱼免疫和抗氧化性能的影响
使用不同CLA含量的日粮经过10周的饲喂后,D1-D4组大黄鱼的脾脏指数随着CLA含量的增加而显著增加,且与对照组相比差异显著(P＜0.05)。但是D3组和D4间无显著差异(P＞0.05)。血清中的溶菌酶和免疫球蛋白M(IgM)活性均有所提高,且与对照组相比差异显著(P＜0.05)。CLA能够提高血清补体C3、C4的活性,D3、D4组与D1组相比差异显著(P＜0.05)。此外,D3组的补体活性显著高于其它三组(P＜0.05),与D1组相比,补体C3、C4活性分别提高了59%和65%。CLA也能够提高血清和组织中的SOD活性。肝脏中的SOD活性最高为202.89 U/ml,约为血清和肌肉SOD活性的1.7倍。D2-D4组的血清和肝脏SOD活性均显著高于D1组(P＜0.05),但是CLA处理组之间差异并不显著(P＞0.05)。
3.日粮CLA水平对大黄鱼肉品质的影响
日粮中添加CLA能够显著影响大黄鱼肌肉的鲜味氨基酸和肌苷酸含量,且随着CLA含量的增加而增加(P＜0.05)。采用微波蒸馏-固相微萃取技术萃取大黄鱼肌肉中的挥发性成分,共检测出54中化合物,其中主要为挥发性醇类和羰基化合物。通过比较发现,各组大黄鱼肌肉挥发性化合物的种类及百分含量没有显著差异(P＞0.05),但是能够提高部分具有特殊气味的醛酮类化合物含量。
4.日粮CLA水平对大黄鱼肝脏PPAR基因表达的影响
通过对大黄鱼肝脏PPARa和PPARγ基因进行克隆,确定了该两种基因的DNA序列,并采用实时荧光定量PCR法对PPARa和PPARγ基因进行了表达水平的分析,结果表明,日粮中添加CLA能够显著提高大黄鱼肝脏中PPARa和PPARγ基因的表达量(P＜0.05)。
Conjugated linoletic acid (CLA) is one of natural unsaturated fatty acids with many physiological functions. It has been testified by experiments on human beings and animals that CLA are characterized for a variety of special functions, including lowing fat content, increasing muscle, anticancer, antiatherosclerosis, enhancing immunity, and improving metabolism of bony tissue. In the study, different levels of CLA were added to diet of large yellow croaker. Effects of various levels of CLA (0, 1, 2, or 4%) on the growth, lipid metabolism, immunity, and muscular properties of large yellow croaker were investigated. Moreover, PPARa and PPARγin liver of large yellow croaker were cloned and expressed, which demonstrated the action mechanism of CLA on lipid metabolism of large yellow croaker in molecular level. The main content and results were listed as follows:
1. Effects of dietary CLA on growth and lipid metaboliam in large yellow croaker
The study was conducted to investigate the effects of dietary conjugated linoleic acid (CLA) on growth, body composition, lipid content, lipogenic enzyme and fatty acid composition of large yellow croaker (Pseudosciaena crocea, R). Four dietary treatments were formulated to contain 0%, 1%, 2% or 4% CLA (instead of fish oil) in diets, respectively. The fish were fed for 10 weeks with ad libitum twice daily. Ten weeks later, as is shown by the results, the specific growth rate (SGR) and condition factor (CF) of large yellow croaker were increased with addition of 1% CLA, but SGR was decreased while high content of CLA (2% & 4%) was added. Meanwhile, little difference was found among the four dietary treatments (P<0.05). Feed conversion ratio (FCR) were not significantly affected by dietary CLA (P>0.05). Hepato-somatic indices (HSI) and viscero-somatic indices (VSI) were both slightly increased in fish fed dietary CLA, but the difference was not significant (P>0.05). With an increasing of CLA concentration, protein, moisture and ash in whole body increased gradually, but the difference was not significant between groups (P>0.05). The dissimilar result was observed in lipid content of liver and flesh. Dietary CLA had no significant effects on lipid content, although there were trends of decreasing in fish fed high CLA levels.
Compared with the control group (D1), the triglyceride (TG) content were significantly decreased, whereas the free fatty acid (FFA) content were significantly increased (P<0.05) in both serum and liver. The TG contents were decreased with the increasing of dietary CLA levels. In serum, notable difference were observed between D3, D4 and D2 (P<0.05). In liver, however, no significant difference was found between treatment groups (D2-D4) (P<0.05). In serum and liver, the contents of FFA in D4 was highest and the difference was marked between D1, D2 and D4 (P<0.05). The content of cholesterol (CHO) and low density lipoprotein - cholesterol (LDL-C) in serum were much lower than D1 (P<0.05), and high density lipoprotein -cholesterol (HDL-C) content in serum was rather higher than D1 (P<0.05). However, no significant difference was observed between treatment groups (P>0.05).
Lipogenic activities in liver of large yellow croaker during the experiment were detected in the study. The activities of hepatic lipase (HL) and lipoprotein lipase (LPL) were decreased significantly by CLA incorporation levels. In both serum and liver, the difference was significant between control and treatment groups, and apparent difference was also found between D3, D4 and D2 (P<0.05). Besides, dietary CLA had diverse influence on the activities of glucose-6-phosphate dehydrogenase (G6PD), fatty acid synthetase (FAS) and malic enzyme (ME). There were no strong effects on G6PD activities both in serum and liver in fish fed with diets containing 1% CLA. However, the activities of G6PD were reduced in fish fed with high CLA levels (2% and 4%), and the difference was significant between D2, D4 and D1 (P<0.05). CLA seemed to decrease the activities of FAS in serum. The FAS activity in serum of D4 was about 16% lower than that of D1, and the difference between D1 and D4 was significant (P<0.05). Similar results were observed in liver, but the difference between four groups was not significant (P>0.05). There was a downward trend for the activities of ME both on serum and liver in fish fed CLA. In serum, the difference between D3, D4 and D1 was significant (P<0.05), and there was no statistically significant between four groups in liver (P>0.05).
Dietary CLA had great influence on fatty acid composition in both muscle and liver (P<0.05). The main fatty acids were C16:0 of saturated fatty acids (SFA), and C18:l n-7 of monounsaturated fatty acids (MUFA). MUFA decreased significantly whereas SFA and polyunsaturated fatty acids (PUFA) showed a clear increase (P<0.05). Concerning the PUFA fraction, a gradual increase of EPA (65% of PUFA) and DHA (63% of PUFA) were observed in both tissues with increasing CLA levels. The fatty acid compositions of muscle and liver reflected the fatty acid composition of the diets. Dietary CLA resulted in the deposition of the cis-9, trans-11 and trans-10, cis-12 CLA isomer in both tissues.
2. Effects of dietary CLA on immunity and antioxidation in large yellow croaker
Effects of dietary CLA supplementation on immune parameters of large yellow croaker were investigated. With the increasing of CLA concentration, the spleen body index (SBI) were increased remarkably (P<0.05), but the difference of D3 and D4 was not significant. Compared with D1, the activities of lysozyme and immunoglobulin M (IgM) were increased significantly (P<0.05). Dietary CLA had great affect on activities of complement components C3, C4 (P<0.05). The activities in D3 were the highest than the other groups (P<0.05), increased by 59% and 65% respectively, compared with D1. The activities of SOD in both serum and tissues were augmented in fish fed CLA supplementation. The SOD activities in liver had reached 202.89 U/ml, about 1.7 fold of that in serum and muscle. In both serum and liver, the SOD activities in treatment groups were higher than control group significantly (P<0.05), and no significant difference were found between treatment groups.
3. Effects of dietary CLA on flesh quality in large yellow croaker
Effects of dietary CLA on amino acid (AA), inosine monphosphate (IMP) and volatile flavor components of large yellow croaker were investigated. With the increasing levels of dietary CLA, essential amino acids and flavour amino acids contents in fillets of large yellow croaker increased, and the difference of flavour AA contents was significantly (P<0.05). IMP content in fillets of large yellow croaker was remarkably increased by supplementing CLA into basal diets (P<0.05).
Volatile flavour compounds from muscle of large yellow croaker were extracted by microwave distillation and solid phase microextraction. The technique of gas chromatography-mass spectrometry was employed to analyze the components, and 54 compounds which mainly contain alcohols, aldehydes and ketones, were identified. By comparing the results of all groups, the percentage profiles of components were not affected by dietary CLA levels, but the contents of aldehydes and ketones with special odor were increased slightly.
4. Effects of dietary CLA on expression of PPAR gene in liver of large yellow croaker
The DNA sequence of PPARa and PPARγgene in liver of large yellow croaker were identified by cloning the two genes. The expression levels of PPARa and PPARγwere analyzed by using the real-time quantitative PCR method. The results showed that dietary CLA had significant influence on PPARa and PPARγexpression (P<0.05). PPARa and PPARγexpression levels were significantly increased compared with D1 (P<0.05).
1.日粮CLA水平对大黄鱼生长性能和脂肪代谢的影响
本试验研究了日粮CLA对大黄鱼生长性能、体成分、脂质组成、脂肪代谢酶活性和脂肪酸成分的影响。在基础日粮中用CLA代替鱼油,添加水平分别为0%、1%、2%和4%(试验D1-D4组)。经过10周的饲喂,结果表明:饲喂1%的CLA能够提高大黄鱼的特定生长率(SGR)和肥满度(CF),饲喂高含量(2%和4%)的CLA则降低了SGR,但是四个试验组之间的差异不显著(P＞0.05)。日粮中添加CLA对大黄鱼的饵料系数(FCR)没有影响,但是肝体比(HSI)和脏体比(VSI)均有所提高,各处理组(D2-D4)之间差异也不显著(P＞0.05)。与对照组(D1)相比,大黄鱼鱼体的水分、粗蛋白和灰分均有所增加,粗脂肪含量有所下降,但是差异不显著(P＞0.05)。随着日粮中CLA水平的增加,大黄鱼肝脏和肌肉中的脂肪含量也有所降低(P＞0.05)。
与D1组相比,CLA能够显著降低血清和肝脏中甘油三酯(TG)的含量(P＜0.05)。血清中的TG含量随着CLA含量的增加显著降低,D2组与D3和D4组相比差异显著(P＜0.05)。在肝脏中,CLA处理组之间差异不显著(P＞0.05)。与此同时,CLA能够显著增加血清和肝脏中游离脂肪酸(FFA)的含量(P＜0.05)。在血清和肝脏中,D4组的FFA含量最高,并且与D1和D2组相比差异显著(P＜0.05)。随着日粮中CLA含量的增加,血清中胆固醇(CHO)和低密度脂蛋白胆固醇(LDL-C)含量也随着降低,CLA处理组与D1组相比有显著差异(P＜0.05)。血清中高密度脂蛋白胆固醇(HDL-C)的含量随着CLA含量的增加而增加。CLA处理组与D1组相比差异显著(P＜0.05),但是D2、D3和D4组之间无显著差异(P＞0.05)。
日粮中添加CLA能够显著降低大黄鱼血清和肝脏中肝脂酶(HL)和脂蛋白脂酶(LPL)的活性。在血清和肝脏中,D2、D3和D4组的HL和LPL活性与D1相比差异显著(P＜0.05),而且D2组与D3和D4相比也具有显著性差异(P＜0.05)。此外,CLA对大黄鱼血清和肝脏中葡萄糖-6-磷酸脱氢酶(G6PD)、脂肪合成酶(FAS)和苹果酸酶(ME)的活性有着不同程度的影响,1%CLA对血清和肝脏中的G6PD活性没有显著影响(P＞0.05)。但是高含量的CLA(2%和4%)能够显著降低血清和肝脏中的G6PD活性,并且与D1组相比差异显著(P＜0.05)。CLA能够显著降低大黄鱼血清中的脂肪酶(FAS)活性,与D1相比,D4组的FAS活性降低了16%,且差异显著(P＜0.05)。与此同时,CLA也能够降低大黄鱼肝脏中的FAS活性,但D1-D4组间差异不显著(P＞0.05)。大黄鱼血清和肝脏中苹果酸酶(ME)的活性随着日粮中CLA含量的增加而降低。在血清中,D3和D4组ME活性与D1相比差异显著(P＜0.05),而D2组与D1组相比差异不显著(P＞0.05)。在肝脏中,D2、D3和D4组之间的ME活性无显著差异(P＞0.05)。
CLA对肝脏和肌肉脂肪酸成分有显著的影响。饱和脂肪酸(SFA)的主要成分是C16:0;单不饱和脂肪酸(MUFA)的主要成分是C18:1 n-9。随着CLA含量的增加,肝脏和肌肉中SFA含量和多不饱和脂肪酸(PUFA)含量也随之增加,而MUFA含量随之降低,并且D1-D4之间差异显著(P＜0.05)。在PUFA中,EPA(C20:5 n-3)和DHA(C22:6 n-3)约占PUFA的65%和63%,并且随着CLA含量的增加而显著增加。此外,肝脏和肌肉中CLA的含量与日粮中CLA含量相同呈等比增加。
2.日粮CLA水平对大黄鱼免疫和抗氧化性能的影响
使用不同CLA含量的日粮经过10周的饲喂后,D1-D4组大黄鱼的脾脏指数随着CLA含量的增加而显著增加,且与对照组相比差异显著(P＜0.05)。但是D3组和D4间无显著差异(P＞0.05)。血清中的溶菌酶和免疫球蛋白M(IgM)活性均有所提高,且与对照组相比差异显著(P＜0.05)。CLA能够提高血清补体C3、C4的活性,D3、D4组与D1组相比差异显著(P＜0.05)。此外,D3组的补体活性显著高于其它三组(P＜0.05),与D1组相比,补体C3、C4活性分别提高了59%和65%。CLA也能够提高血清和组织中的SOD活性。肝脏中的SOD活性最高为202.89 U/ml,约为血清和肌肉SOD活性的1.7倍。D2-D4组的血清和肝脏SOD活性均显著高于D1组(P＜0.05),但是CLA处理组之间差异并不显著(P＞0.05)。
3.日粮CLA水平对大黄鱼肉品质的影响
日粮中添加CLA能够显著影响大黄鱼肌肉的鲜味氨基酸和肌苷酸含量,且随着CLA含量的增加而增加(P＜0.05)。采用微波蒸馏-固相微萃取技术萃取大黄鱼肌肉中的挥发性成分,共检测出54中化合物,其中主要为挥发性醇类和羰基化合物。通过比较发现,各组大黄鱼肌肉挥发性化合物的种类及百分含量没有显著差异(P＞0.05),但是能够提高部分具有特殊气味的醛酮类化合物含量。
4.日粮CLA水平对大黄鱼肝脏PPAR基因表达的影响
通过对大黄鱼肝脏PPARa和PPARγ基因进行克隆,确定了该两种基因的DNA序列,并采用实时荧光定量PCR法对PPARa和PPARγ基因进行了表达水平的分析,结果表明,日粮中添加CLA能够显著提高大黄鱼肝脏中PPARa和PPARγ基因的表达量(P＜0.05)。
Conjugated linoletic acid (CLA) is one of natural unsaturated fatty acids with many physiological functions. It has been testified by experiments on human beings and animals that CLA are characterized for a variety of special functions, including lowing fat content, increasing muscle, anticancer, antiatherosclerosis, enhancing immunity, and improving metabolism of bony tissue. In the study, different levels of CLA were added to diet of large yellow croaker. Effects of various levels of CLA (0, 1, 2, or 4%) on the growth, lipid metabolism, immunity, and muscular properties of large yellow croaker were investigated. Moreover, PPARa and PPARγin liver of large yellow croaker were cloned and expressed, which demonstrated the action mechanism of CLA on lipid metabolism of large yellow croaker in molecular level. The main content and results were listed as follows:
1. Effects of dietary CLA on growth and lipid metaboliam in large yellow croaker
The study was conducted to investigate the effects of dietary conjugated linoleic acid (CLA) on growth, body composition, lipid content, lipogenic enzyme and fatty acid composition of large yellow croaker (Pseudosciaena crocea, R). Four dietary treatments were formulated to contain 0%, 1%, 2% or 4% CLA (instead of fish oil) in diets, respectively. The fish were fed for 10 weeks with ad libitum twice daily. Ten weeks later, as is shown by the results, the specific growth rate (SGR) and condition factor (CF) of large yellow croaker were increased with addition of 1% CLA, but SGR was decreased while high content of CLA (2% & 4%) was added. Meanwhile, little difference was found among the four dietary treatments (P<0.05). Feed conversion ratio (FCR) were not significantly affected by dietary CLA (P>0.05). Hepato-somatic indices (HSI) and viscero-somatic indices (VSI) were both slightly increased in fish fed dietary CLA, but the difference was not significant (P>0.05). With an increasing of CLA concentration, protein, moisture and ash in whole body increased gradually, but the difference was not significant between groups (P>0.05). The dissimilar result was observed in lipid content of liver and flesh. Dietary CLA had no significant effects on lipid content, although there were trends of decreasing in fish fed high CLA levels.
Compared with the control group (D1), the triglyceride (TG) content were significantly decreased, whereas the free fatty acid (FFA) content were significantly increased (P<0.05) in both serum and liver. The TG contents were decreased with the increasing of dietary CLA levels. In serum, notable difference were observed between D3, D4 and D2 (P<0.05). In liver, however, no significant difference was found between treatment groups (D2-D4) (P<0.05). In serum and liver, the contents of FFA in D4 was highest and the difference was marked between D1, D2 and D4 (P<0.05). The content of cholesterol (CHO) and low density lipoprotein - cholesterol (LDL-C) in serum were much lower than D1 (P<0.05), and high density lipoprotein -cholesterol (HDL-C) content in serum was rather higher than D1 (P<0.05). However, no significant difference was observed between treatment groups (P>0.05).
Lipogenic activities in liver of large yellow croaker during the experiment were detected in the study. The activities of hepatic lipase (HL) and lipoprotein lipase (LPL) were decreased significantly by CLA incorporation levels. In both serum and liver, the difference was significant between control and treatment groups, and apparent difference was also found between D3, D4 and D2 (P<0.05). Besides, dietary CLA had diverse influence on the activities of glucose-6-phosphate dehydrogenase (G6PD), fatty acid synthetase (FAS) and malic enzyme (ME). There were no strong effects on G6PD activities both in serum and liver in fish fed with diets containing 1% CLA. However, the activities of G6PD were reduced in fish fed with high CLA levels (2% and 4%), and the difference was significant between D2, D4 and D1 (P<0.05). CLA seemed to decrease the activities of FAS in serum. The FAS activity in serum of D4 was about 16% lower than that of D1, and the difference between D1 and D4 was significant (P<0.05). Similar results were observed in liver, but the difference between four groups was not significant (P>0.05). There was a downward trend for the activities of ME both on serum and liver in fish fed CLA. In serum, the difference between D3, D4 and D1 was significant (P<0.05), and there was no statistically significant between four groups in liver (P>0.05).
Dietary CLA had great influence on fatty acid composition in both muscle and liver (P<0.05). The main fatty acids were C16:0 of saturated fatty acids (SFA), and C18:l n-7 of monounsaturated fatty acids (MUFA). MUFA decreased significantly whereas SFA and polyunsaturated fatty acids (PUFA) showed a clear increase (P<0.05). Concerning the PUFA fraction, a gradual increase of EPA (65% of PUFA) and DHA (63% of PUFA) were observed in both tissues with increasing CLA levels. The fatty acid compositions of muscle and liver reflected the fatty acid composition of the diets. Dietary CLA resulted in the deposition of the cis-9, trans-11 and trans-10, cis-12 CLA isomer in both tissues.
2. Effects of dietary CLA on immunity and antioxidation in large yellow croaker
Effects of dietary CLA supplementation on immune parameters of large yellow croaker were investigated. With the increasing of CLA concentration, the spleen body index (SBI) were increased remarkably (P<0.05), but the difference of D3 and D4 was not significant. Compared with D1, the activities of lysozyme and immunoglobulin M (IgM) were increased significantly (P<0.05). Dietary CLA had great affect on activities of complement components C3, C4 (P<0.05). The activities in D3 were the highest than the other groups (P<0.05), increased by 59% and 65% respectively, compared with D1. The activities of SOD in both serum and tissues were augmented in fish fed CLA supplementation. The SOD activities in liver had reached 202.89 U/ml, about 1.7 fold of that in serum and muscle. In both serum and liver, the SOD activities in treatment groups were higher than control group significantly (P<0.05), and no significant difference were found between treatment groups.
3. Effects of dietary CLA on flesh quality in large yellow croaker
Effects of dietary CLA on amino acid (AA), inosine monphosphate (IMP) and volatile flavor components of large yellow croaker were investigated. With the increasing levels of dietary CLA, essential amino acids and flavour amino acids contents in fillets of large yellow croaker increased, and the difference of flavour AA contents was significantly (P<0.05). IMP content in fillets of large yellow croaker was remarkably increased by supplementing CLA into basal diets (P<0.05).
Volatile flavour compounds from muscle of large yellow croaker were extracted by microwave distillation and solid phase microextraction. The technique of gas chromatography-mass spectrometry was employed to analyze the components, and 54 compounds which mainly contain alcohols, aldehydes and ketones, were identified. By comparing the results of all groups, the percentage profiles of components were not affected by dietary CLA levels, but the contents of aldehydes and ketones with special odor were increased slightly.
4. Effects of dietary CLA on expression of PPAR gene in liver of large yellow croaker
The DNA sequence of PPARa and PPARγgene in liver of large yellow croaker were identified by cloning the two genes. The expression levels of PPARa and PPARγwere analyzed by using the real-time quantitative PCR method. The results showed that dietary CLA had significant influence on PPARa and PPARγexpression (P<0.05). PPARa and PPARγexpression levels were significantly increased compared with D1 (P<0.05).
引文
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