【摘要】：本試驗(yàn)通過體外試驗(yàn)和體內(nèi)試驗(yàn)對(duì)槲皮素的抗氧化作用進(jìn)行研究。體外試驗(yàn)以維生素C為陽性對(duì)照,分別從清除自由基、保護(hù)生物大分子及總還原力3方面綜合評(píng)價(jià)槲皮素的體外抗氧化能力;體內(nèi)試驗(yàn)采用單因素完全隨機(jī)試驗(yàn)設(shè)計(jì),將240只健康、體重和產(chǎn)蛋率相近的37周齡海賽蛋雞隨機(jī)分4組,每組6個(gè)重復(fù),每個(gè)重復(fù)10只雞,對(duì)照組飼喂玉米-豆粕型基礎(chǔ)飼糧,試驗(yàn)I、II、III組分別在基礎(chǔ)飼糧中添加0.02%、0.04%、0.06%槲皮素,研究槲皮素對(duì)海賽蛋雞抗氧化指標(biāo)和生產(chǎn)性能的影響,同時(shí)探討槲皮素作為飼糧添加劑的最適添加量。1.槲皮素對(duì)自由基的清除作用分別采用鄰苯三酚自氧化法、水楊酸法和1,1-二苯基-2-三硝基苯肼(1,1-Diphenyl-2-picrylhydrazyl,DPPH)法探討槲皮素對(duì)超氧陰離子、羥自由基和DPPH的清除作用,結(jié)果發(fā)現(xiàn),槲皮素對(duì)O2-清除作用隨濃度增加先升高后降低(P0.01),40 mg/L清除率最大(31.93%),但清除作用小于維生素C;槲皮素對(duì)·OH清除作用也隨濃度的增加先升高再降低(P0.01),300 mg/L清除率最大(61.79%),IC50=218.4 mg/L;槲皮素對(duì)DPPH清除作用隨濃度增加不斷升高,IC50=1.47 mg/L,清除·OH和DPPH作用均在低濃度時(shí)大于維生素C,高濃度時(shí)小于維生素C,槲皮素對(duì)3種自由基的清除作用為DPPH·OHO2-。2.槲皮素對(duì)卵黃脂蛋白過氧化的抑制作用采用Fe2+誘發(fā)卵黃脂蛋白多不飽和脂肪酸體系研究槲皮素對(duì)卵黃脂蛋白過氧化的抑制作用,結(jié)果發(fā)現(xiàn),槲皮素對(duì)卵黃脂蛋白過氧化的抑制作用隨濃度升高不斷增大(P0.01),IC50=11.89 mg/L,對(duì)脂質(zhì)過氧化的抑制作用高于維生素C。3.槲皮素對(duì)紅細(xì)胞溶血和線粒體腫脹的抑制作用分別采用H2O2誘導(dǎo)紅細(xì)胞溶血,VC-Fe2+誘導(dǎo)線粒體膨脹體系研究槲皮素對(duì)紅細(xì)胞和線粒體的保護(hù)作用,結(jié)果發(fā)現(xiàn),槲皮素在0.05~1.25 mg/L時(shí)對(duì)紅細(xì)胞溶血抑制作用隨濃度的升高極顯著增加(P0.01),但濃度過高時(shí)(31.25 mg/L)抑制作用反而下降,IC50=0.16 mg/L,對(duì)紅細(xì)胞溶血抑制作用強(qiáng)于維生素C;槲皮素對(duì)線粒體膨脹抑制作用隨濃度增加不斷增大,根據(jù)60 min時(shí)線粒體的膨脹程度可知,試驗(yàn)組線粒體的完整程度極顯著高于模型組(P0.01)。4.槲皮素的總還原力采用普魯士藍(lán)法對(duì)槲皮素總還原力進(jìn)行測(cè)定,結(jié)果表明,槲皮素總還原力隨濃度增加而增大(P0.01),經(jīng)計(jì)算濃度(x)與還原力(y)的關(guān)系為y=-0.0001x2+0.0263x+0.0391,R2=0.9992;根據(jù)回歸方程可知,槲皮素的總還原力小于維生素C。5.槲皮素對(duì)蛋雞抗氧化指標(biāo)的影響體內(nèi)試驗(yàn)結(jié)果表明,與對(duì)照組相比,3水平槲皮素組蛋雞血清總抗氧化能力均極顯著升高(P0.01);0.04%槲皮素組血清丙二醛含量顯著降低(P0.05),谷胱甘肽過氧化物酶和超氧化物歧化酶活性極顯著升高(P0.01),0.06%槲皮素組血清超氧化物歧化酶活性顯著升高(P0.05);3水平槲皮素組肝臟超氧化物歧化酶活性顯著升高(P0.05,P0.01,P0.05)。6.槲皮素對(duì)蛋雞生產(chǎn)性能的影響同對(duì)照組相比,0.02%和0.04%槲皮素組產(chǎn)蛋率分別極顯著提高4.96%和8.11%(P0.01),料蛋比分別顯著下降6.55%和11.72%(P0.05,P0.01)。綜上所述,槲皮素在一定濃度范圍內(nèi)具有較強(qiáng)的抗氧化作用,且可以提高37~45周齡蛋雞的抗氧化能力和生產(chǎn)性能,槲皮素用于提高蛋雞生產(chǎn)性能的最適添加量為0.04%。
[Abstract]:The antioxidant activity of quercetin was studied in vitro and in vivo. Vitamin C was used as the positive control in vitro to evaluate the antioxidant activity of quercetin in vitro from scavenging free radicals, protecting biological macromolecules and total reducing power. In vivo, the single factor complete randomized trial design was used to evaluate the antioxidant activity of quercetin. Healthy 37-week-old Haisai layers were randomly divided into 4 groups with 6 replicates in each group and 10 chickens in each replicate. The control group was fed with corn-soybean meal basal diet. The experimental group I, II and III were fed with 0.02%, 0.04% and 0.06% quercetin respectively in the basal diet. The scavenging effects of quercetin on free radicals were studied by pyrogallol autoxidation, salicylic acid and 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) methods. The scavenging effects of quercetin on superoxide anions, hydroxyl radicals and DPPH were investigated. The scavenging effect of quercetin on O2 - increased first and then decreased with the increase of concentration (P 0.01). The scavenging effect of quercetin on DPPH was the highest (31.93%) at 40 mg/L, but less than that of vitamin C. The scavenging effect of quercetin on OH increased first and then decreased with the increase of concentration (P 0.01). The scavenging effect of quercetin on DPPH was the highest (61.79%) at 300 mg/L and IC50 = 218.4 mg/L at 300 mg/L. IC50 = 1.47 mg/L, clearance of OH and DPPH were higher than that of vitamin C at low concentration and lower than that of vitamin C at high concentration. The scavenging effect of quercetin on three free radicals was DPPH OHO2 -.2. Inhibitory effect of quercetin on yolk lipoprotein peroxidation was studied by Fe2+ induced yolk lipoprotein polyunsaturated fatty acid system. Inhibitory effect of quercetin on the peroxidation of yolk lipoprotein was found to be increasing with the increase of concentration (P 0.01), IC50 = 11.89 mg/L. Inhibitory effect of quercetin on lipid peroxidation was higher than that of vitamin C.3. Inhibitory effect of quercetin on red blood cell hemolysis and mitochondrial swelling induced by H2O2 respectively. The protective effects of quercetin on erythrocyte and mitochondria were studied by cytolysis and mitochondrial dilatation induced by VC-Fe2+. The results showed that the inhibition of quercetin on erythrocyte hemolysis increased significantly with the increase of concentration (P 0.01), but the inhibition decreased when the concentration was too high (31.25 mg/L), and the IC50 = 0.16 mg/L. The inhibitory effect of quercetin on mitochondrial swelling was stronger than that of vitamin C, and the inhibitory effect of quercetin on mitochondrial swelling increased with the increase of concentration. According to the degree of mitochondrial swelling at 60 min, the mitochondrial integrity of the experimental group was significantly higher than that of the model group (P 0.01). 4. The total reducing power of quercetin was determined by Prussian blue method. The results showed that the total reducing power of quercetin increased with the increase of the concentration (P 0.01), and the relationship between the calculated concentration (x) and the reducing power (y) was y = - 0.0001x2 + 0.0263x + 0.0391, R2 = 0.9992. According to the regression equation, the total reducing power of quercetin was less than that of vitamin C.5. The total antioxidant capacity of serum in quercetin group was significantly increased (P 0.01), the content of malondialdehyde (P 0.05), the activity of glutathione peroxidase and superoxide dismutase (P 0.01), and the activity of superoxide dismutase (SOD) in quercetin group was significantly increased (P 0.05) at 0.04% and 0.06% respectively. The effect of quercetin on the performance of laying hens was significantly increased by 4.96% and 8.11% in 0.02% and 0.04% quercetin groups, respectively (P 0.01), and the feed-egg ratio decreased by 6.55% and 11.72% respectively (P 0.05, P 0.01). It has strong antioxidant effect in the periphery and can improve the antioxidant ability and production performance of 37-45 weeks old laying hens. The optimum addition of quercetin to improve the production performance of laying hens is 0.04%.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S831.5

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