日粮中添加水飞蓟素对草鱼脂质代谢的影响及其机制研究

发布时间：2018-03-23 22:12

  本文选题：水飞蓟素　切入点：日粮脂肪　出处：《西北农林科技大学》2017年博士论文

【摘要】：水飞蓟素是一种黄酮类的植物提取物,从紫花水飞蓟(Silybum marianum L.)的干燥果实中提取,主要由四种黄酮类化合物组成,包含水飞蓟宾A和B,异水飞蓟宾A和B,水飞蓟宁和水飞蓟亭。在哺乳动物上,水飞蓟素具有保护肝脏、抗氧化抗炎作用,降血脂和降低内脏脂肪沉积等作用,而有关水飞蓟素对鱼类脂质代谢的研究鲜有报道。在生产实践中,饲料中油脂的过量添加或氧化油脂的使用,给养殖鱼类带来诸多问题,如内脏脂肪过度蓄积,引起脂肪肝、腹腔脂肪过度沉积、抗应激力下降、肉质品质和风味变化等,这已经成为困扰和危害养殖业健康发展的问题之一。针对当前面临的问题,本研究探讨了不同脂肪或氧化油脂水平日粮中添加水飞蓟素对鱼体生长、体组成、脂质代谢和健康的影响。与此同时,利用离体细胞平台研究,探讨了水飞蓟素对肝细胞和脂肪细胞分化过程中脂质蓄积抑制的作用机制。主要试验结果和结论如下:1、日粮添加水飞蓟素能够抑制高脂日粮诱导的肝胰脏和腹腔脂肪组织脂质的蓄积,并提高草鱼抗氧化能力。采用2×3双因素试验设计方法,设计4%和8%脂肪水平(分别是low lipid,LL,和high lipid,HL)和0,100,200mg kg-1水飞蓟素水平(SM0,SM100,SM200)的6种等氮等能的半纯化日粮,饲喂草鱼(初均重27.43±0.17克)82天,每个处理组4个重复。结果显示日粮中添加水飞蓟素显著提高鱼体的增重率、特定生长率和蛋白质效率,降低饲料系数。水飞蓟素能够促进肝胰脏脂质分解(HSL)和β-氧化(CPT1)相关基因表达上调,抑制高脂日粮诱导的肝胰脏脂肪沉积。水飞蓟素降低高脂日粮诱导的腹腔脂肪蓄积,通过下调脂质合成相关基因的表达。水飞蓟素可降低高脂日粮诱导的MDA含量,提高了抗氧化能力,有利于改善鱼体健康。2、水飞蓟素可以缓解氧化豆油对鱼体生长的副作用,并改善氧化油脂诱导鱼体脂质过氧化反应,降低鱼体氧化应激。采用2×3双因素试验设计方法,设计2.5%和6.5%氧化豆油水平(分别Low oxidized soybean oil level,High oxidized soybean oil level,简写为LO和HO)和3个水飞蓟素浓度(0,100 mg kg-1和200 mg kg-1,简写为SM0,SM100,SM200)的6个处理组,并分别设置新鲜低豆油组(2.5%Low fresh oil,LF)和新鲜高豆油组(6.5%High fresh oil,HF)作为低氧化油脂组和高氧化豆油组的正常对照组,配制8种等氮(粗蛋白30%)的半纯化日粮,饲喂草鱼(初均重60.01±0.77 g)80天,每个处理组3个重复。鱼体的增重率、特定生长率、日摄食率、饵料系数、成活率在所有试验处理组间均无显著性差异,但在高氧化豆油日粮中添加水飞蓟素后鱼体增重率比HOSM0组平均提高了8.41-8.80%,低氧化豆油中添加水飞蓟素后增重率比LOSM0组平均增加了10.16-19.77%,显示水飞蓟素可缓解氧化油脂对鱼体生长的副作用。水飞蓟素可降低氧化油脂诱导鱼体MDA含量,并提高抗氧化能力参数(CAT、GSH-Px活性),表明水飞蓟素能够提高机体抗氧化能力,有利于提高鱼体健康。3、水飞蓟素能够抑制油酸诱导草鱼肝细胞脂质蓄积,并提高肝细胞抗氧化能力。体外培养草鱼肝细胞,用400μmol L-1油酸对其进行蓄脂诱导的同时,采用不同质量浓度的水飞蓟素处理24h,检测了肝细胞活力、脂质蓄积状况、抗氧化指标和脂质代谢相关基因表达状况。结果显示水飞蓟素质量浓度在100μg mL-1时能够显著抑制肝细胞脂质蓄积,下调脂质合成基因(FAS和SCD1)表达,并能够显著提高肝细胞GSH含量。4、水飞蓟素能够抑制草鱼脂肪细胞分化,并提高脂肪细胞的抗氧化能力。利用离体培养草鱼脂肪细胞的方法,采用不同浓度水飞蓟素(50和100μg mL-1)持续处理分化的前体脂肪细胞6天。水飞蓟素100μg mL-1显著抑制了脂肪细胞甘油三酯含量,并抑制脂肪细胞分化相关的基因PPARγ,C/EBPα,SREBP1c,FAS,SCD1和LPL的m RNA表达以及PPARγ蛋白表达水平。水飞蓟素降低分化的脂肪细胞ROS生成和GSH含量,发挥抗氧化作用。综上所述,水飞蓟素能够抑制高脂日粮诱导的肝胰脏和腹腔脂肪组织脂质过度蓄积,可以缓解氧化油脂对鱼体的副作用,降低鱼体氧化应激,提高鱼体抗氧化能力,有利于改善鱼体健康。
[Abstract]:Silymarin is a botanical extract flavonoids, from Silybum marianum (Silybum marianum L.) extracted from dried fruit, mainly composed of four flavonoids, including silybin A and B, isosilybin A and B, silybin and isosilybin. In mammals. Silymarin can protect the liver antioxidant, anti-inflammatory effect, lowering blood lipid and reducing visceral fat deposition, and the effects of silymarin on fish lipid metabolism has not been studied. In practice, the use of fat in diet or excessive addition of oxidized oil, brought many problems to the fish, such as excessive accumulation of visceral fat, the fatty liver caused by excessive deposition, abdominal fat, decreased stress resistance, meat quality and flavor changes, it has become one of the problems and harm the healthy development of aquaculture problems. Aiming at the current problems, in this study. Milk thistle different fat or lipid peroxidation levels in the diets of fish growth hormone, body composition, lipid metabolism and health. At the same time, in vitro cell research platform use, explore the mechanism of silymarin on lipid and fatty liver cells during cell differentiation by repressing. The main results and conclusions are as follows 1, accumulation of milk thistle dietary peptide can inhibit the high fat diet induced liver and abdominal adipose tissue lipid, and improve the antioxidant capacity of grass carp. By 2 * 3 double factor test design method, design 4% and 8% lipid levels (low lipid, LL, and high lipid, and HL) 0100200mg kg-1 of silymarin (SM0, SM100, SM200 level) semi purified diets of 6 kinds of nitrogen and energy, with grass carp (initial average weight 27.43 + 0.17 g) for 82 days, each treatment consisted of 4 replicates. The results showed that with the addition of water of dietary silymarin significantly improved weight The weight gain rate, specific growth rate and protein efficiency, reduce the feed coefficient. Silymarin can promote liver lipid decomposition (HSL) and beta oxidation (CPT1) genes is up-regulated, liver and pancreas fat deposition inhibition induced by high fat diet. Silymarin reduced high fat diet induced peritoneal fat accumulation, by down regulating the expression of lipogenic genes. Silymarin could reduce the content of MDA induced by high fat diet, improve the antioxidant capacity, to improve the health of fish.2, silymarin can alleviate the side effects of oxidized soybean oil fish growth, and improve lipid peroxidation lipid peroxidation induced by fish. Fish reduce oxidative stress. Using 2 * 3 double factor test design method, the design of the 2.5% and 6.5% levels (respectively Low oxidized oxidation of soybean oil soybean oil level, High oxidized soybean oil level, abbreviated as LO and HO) and 3 (the concentration of Silymarin 0100 mg kg-1 and 200 mg kg-1, abbreviated as SM0, SM100, SM200) of the 6 groups, and were set low fresh soybean oil group (2.5%Low fresh oil, LF) and fresh high soybean oil group (6.5%High fresh oil, HF) as normal control group and high fat low oxidation oxidation of soybean oil were prepared. 8 isonitrogenous (30% crude protein) semi purified diets were fed, grass carp (initial average weight 60.01 + 0.77 g) for 80 days, each treatment consisted of 3 replications. The body weight gain rate, specific growth rate, feeding rate, feed coefficient, the survival rate of all treatments showed no significant difference, but the silymarin fish growth rate than the HOSM0 group increased 8.41-8.80% after adding in high oxidized soybean oil diets, low oxidized soybean oil added silymarin after weight gain rate than the LOSM0 group increased by an average of 10.16-19.77%, showed that silymarin can alleviate the side effects of fish oil on oxidation the growth of silymarin can be reduced. The fish oil and the content of MDA induced oxidation, improve antioxidant capacity parameters (CAT, GSH-Px, activity) showed that silymarin can improve the antioxidant capacity, help to improve the health of fish.3, silymarin can inhibit lipid accumulation in liver cells induced by oleic acid in grass carp, and improve the antioxidant ability. Grass carp liver cells liver cells cultured in vitro. Storage fat induced at the same time with 400 mol L-1 oleic acid with different concentrations of silymarin treatment 24h, detection of liver cell viability, lipid accumulation, expression index and lipid metabolism related genes. The results showed that antioxidant silymarin concentration can significantly inhibit hepatocyte lipid accumulation in 100 g mL-1, downregulation of lipogenic gene expression (FAS and SCD1), and can significantly improve the content of GSH in liver cells.4, silymarin can inhibit grass carp adipocyte differentiation, and improve the anti fat cells Oxidation ability. Using the in vitro culture of grass carp fat cells, with different concentrations of silymarin (50 g and 100 mL-1) for treatment of differentiation of preadipocytes in 6 days. Silymarin 100 g mL-1 significantly inhibited adipocyte triglyceride content and inhibit the differentiation related gene PPAR gamma, fat cells SREBP1c, FAS, C/EBP alpha, SCD1 LPL and m RNA expression and PPAR expression on protein level. Silymarin reduced fat cell formation and differentiation of ROS GSH content, exert an antioxidant effect. In summary, silymarin can inhibit the high fat diet induced liver and abdominal adipose tissue lipid accumulation, can alleviate the side effects of oxidized oil on the fish, the fish body to reduce oxidative stress, enhance the antioxidant ability of fish, is conducive to improving the health of fish.

【学位授予单位】：西北农林科技大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S965.112
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本文编号：1655416