本文選題：水飛薊素　切入點：日糧脂肪　出處：《西北農(nóng)林科技大學(xué)》2017年博士論文

【摘要】：水飛薊素是一種黃酮類的植物提取物,從紫花水飛薊(Silybum marianum L.)的干燥果實中提取,主要由四種黃酮類化合物組成,包含水飛薊賓A和B,異水飛薊賓A和B,水飛薊寧和水飛薊亭。在哺乳動物上,水飛薊素具有保護肝臟、抗氧化抗炎作用,降血脂和降低內(nèi)臟脂肪沉積等作用,而有關(guān)水飛薊素對魚類脂質(zhì)代謝的研究鮮有報道。在生產(chǎn)實踐中,飼料中油脂的過量添加或氧化油脂的使用,給養(yǎng)殖魚類帶來諸多問題,如內(nèi)臟脂肪過度蓄積,引起脂肪肝、腹腔脂肪過度沉積、抗應(yīng)激力下降、肉質(zhì)品質(zhì)和風(fēng)味變化等,這已經(jīng)成為困擾和危害養(yǎng)殖業(yè)健康發(fā)展的問題之一。針對當前面臨的問題,本研究探討了不同脂肪或氧化油脂水平日糧中添加水飛薊素對魚體生長、體組成、脂質(zhì)代謝和健康的影響。與此同時,利用離體細胞平臺研究,探討了水飛薊素對肝細胞和脂肪細胞分化過程中脂質(zhì)蓄積抑制的作用機制。主要試驗結(jié)果和結(jié)論如下:1、日糧添加水飛薊素能夠抑制高脂日糧誘導(dǎo)的肝胰臟和腹腔脂肪組織脂質(zhì)的蓄積,并提高草魚抗氧化能力。采用2×3雙因素試驗設(shè)計方法,設(shè)計4%和8%脂肪水平(分別是low lipid,LL,和high lipid,HL)和0,100,200mg kg-1水飛薊素水平(SM0,SM100,SM200)的6種等氮等能的半純化日糧,飼喂草魚(初均重27.43±0.17克)82天,每個處理組4個重復(fù)。結(jié)果顯示日糧中添加水飛薊素顯著提高魚體的增重率、特定生長率和蛋白質(zhì)效率,降低飼料系數(shù)。水飛薊素能夠促進肝胰臟脂質(zhì)分解(HSL)和β-氧化(CPT1)相關(guān)基因表達上調(diào),抑制高脂日糧誘導(dǎo)的肝胰臟脂肪沉積。水飛薊素降低高脂日糧誘導(dǎo)的腹腔脂肪蓄積,通過下調(diào)脂質(zhì)合成相關(guān)基因的表達。水飛薊素可降低高脂日糧誘導(dǎo)的MDA含量,提高了抗氧化能力,有利于改善魚體健康。2、水飛薊素可以緩解氧化豆油對魚體生長的副作用,并改善氧化油脂誘導(dǎo)魚體脂質(zhì)過氧化反應(yīng),降低魚體氧化應(yīng)激。采用2×3雙因素試驗設(shè)計方法,設(shè)計2.5%和6.5%氧化豆油水平(分別Low oxidized soybean oil level,High oxidized soybean oil level,簡寫為LO和HO)和3個水飛薊素濃度(0,100 mg kg-1和200 mg kg-1,簡寫為SM0,SM100,SM200)的6個處理組,并分別設(shè)置新鮮低豆油組(2.5%Low fresh oil,LF)和新鮮高豆油組(6.5%High fresh oil,HF)作為低氧化油脂組和高氧化豆油組的正常對照組,配制8種等氮(粗蛋白30%)的半純化日糧,飼喂草魚(初均重60.01±0.77 g)80天,每個處理組3個重復(fù)。魚體的增重率、特定生長率、日攝食率、餌料系數(shù)、成活率在所有試驗處理組間均無顯著性差異,但在高氧化豆油日糧中添加水飛薊素后魚體增重率比HOSM0組平均提高了8.41-8.80%,低氧化豆油中添加水飛薊素后增重率比LOSM0組平均增加了10.16-19.77%,顯示水飛薊素可緩解氧化油脂對魚體生長的副作用。水飛薊素可降低氧化油脂誘導(dǎo)魚體MDA含量,并提高抗氧化能力參數(shù)(CAT、GSH-Px活性),表明水飛薊素能夠提高機體抗氧化能力,有利于提高魚體健康。3、水飛薊素能夠抑制油酸誘導(dǎo)草魚肝細胞脂質(zhì)蓄積,并提高肝細胞抗氧化能力。體外培養(yǎng)草魚肝細胞,用400μmol L-1油酸對其進行蓄脂誘導(dǎo)的同時,采用不同質(zhì)量濃度的水飛薊素處理24h,檢測了肝細胞活力、脂質(zhì)蓄積狀況、抗氧化指標和脂質(zhì)代謝相關(guān)基因表達狀況。結(jié)果顯示水飛薊素質(zhì)量濃度在100μg mL-1時能夠顯著抑制肝細胞脂質(zhì)蓄積,下調(diào)脂質(zhì)合成基因(FAS和SCD1)表達,并能夠顯著提高肝細胞GSH含量。4、水飛薊素能夠抑制草魚脂肪細胞分化,并提高脂肪細胞的抗氧化能力。利用離體培養(yǎng)草魚脂肪細胞的方法,采用不同濃度水飛薊素(50和100μg mL-1)持續(xù)處理分化的前體脂肪細胞6天。水飛薊素100μg mL-1顯著抑制了脂肪細胞甘油三酯含量,并抑制脂肪細胞分化相關(guān)的基因PPARγ,C/EBPα,SREBP1c,FAS,SCD1和LPL的m RNA表達以及PPARγ蛋白表達水平。水飛薊素降低分化的脂肪細胞ROS生成和GSH含量,發(fā)揮抗氧化作用。綜上所述,水飛薊素能夠抑制高脂日糧誘導(dǎo)的肝胰臟和腹腔脂肪組織脂質(zhì)過度蓄積,可以緩解氧化油脂對魚體的副作用,降低魚體氧化應(yīng)激,提高魚體抗氧化能力,有利于改善魚體健康。
[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.

【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：S965.112
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本文編號：1655416