本文選題：運輸應激 + 大鼠��； 參考：《華中農業(yè)大學》2014年碩士論文

【摘要】：動物的運輸是畜牧生產中一個重要的環(huán)節(jié),運輸應激對動物生產性能、健康狀況等產生一系列負面影響。家畜在運輸應激后使機體內氧化和抗氧化失衡,導致機體內發(fā)生氧化應激。本研究旨在評價抗氧化劑維生素C、維生素E、α-硫辛酸、槲皮素和止痢草油的抗氧化能力和特性,并通過建立大鼠運輸應激模型,比較這5種抗氧化劑緩解大鼠運輸應激的效果,為家畜運輸應激的營養(yǎng)調控提供理論依據。主要研究內容和結果如下： 1、不同抗氧化劑體外抗氧化效果評價及特性研究。利用DPPH法測定維生素C、維生素E、槲皮素和止痢草油清除DPPH自由基的能力,結果表明抗氧化劑清除DPPH自由基的IC50如下：維生素C為0.0059mg/ml,維生素E為0.0249mg/ml,槲皮素為0.0051mg/ml,止痢草油為0.0687mg/ml,這4種抗氧化劑對DPPH自由基的清除能力大小順序為：槲皮素維生素C維生素E止痢草油。通過建立大鼠肝勻漿脂質自氧化模型,測定維生素C、a-硫辛酸、維生素E、槲皮素和止痢草油的脂質自氧化抑制率,各抗氧化劑的IC50如下：維生素C為0.255mg/ml,維生素E為0.0052mg/ml,α-硫辛酸為2.06mg/ml,槲皮素為0.0058mg/ml,止痢草油為0.067mg/ml,抑制脂質自氧化的高低順序為：維生素E槲皮素止痢草油維生素Cα-硫辛酸。 2、抗氧化劑對運輸應激大鼠的抗應激效果的研究。試驗選擇7-8周齡SPF級雄性Wistar大鼠70只,隨機分成7組,每組10個重復,每個重復1只大鼠。其中非應激對照組和應激對照組均飼喂基礎日糧,5個日糧處理組在基礎日糧中分別添加1.5g/kg維生素C、2g/kg α-硫辛酸、1g/kg維生素E、1g/kg槲皮素、100mg/kg止痢草油。試驗期28d,試驗期結束后,應激組大鼠根據運輸應激模擬條件(大鼠置于35℃恒溫搖床中,60r/min,2h)進行處理,運輸應激處理結束后乙醚麻醉并屠宰大鼠,收集樣品。測定血清激素水平、血清和肝臟中抗氧化系統指標、空腸組織形態(tài)結構及空腸mRNA表達量。結果顯示： 1)添加止痢草油能顯著增加大鼠體重、日增重和日采食量(P0.05),添加維生素E顯著增加大鼠體重(P0.05),添加槲皮素可以顯著增加大鼠的平均日采食量(P0.05)。 2)運輸導致大鼠體重顯著下降,體溫顯著升高(P0.05)。添加各抗氧化劑后,體重失重與應激對照組無顯著差異(P0.05),運輸后維生素C組大鼠體溫顯著高于其他各組(P0.05)。 3)運輸應激處理后應激對照組大鼠血清中皮質醇和去甲腎上腺素水平均顯著升高(P0.05)。與應激對照組相比,日糧添加維生素E顯著降低了去甲腎上腺素水平(P0.05)。 4)與非應激組相比,應激對照組的血清T-AOC顯著降低(P0.05),MDA水平顯著升高(P0.05),SOD和GSH-Px活力均顯著降低(P0.05)；添加抗氧化劑后,各抗氧化劑組與應激對照組相比,MDA水平顯著降低(P0.05),SOD活力顯著升高(P0.05),維生素C、維生素E與止痢草油組T-AOC顯著升高,維生素E、槲皮素、a-硫辛酸和止痢草油組GSH-Px活力均顯著升高(P0.05)。 5)抗氧化劑對運輸應激大鼠肝臟抗氧化功能的影響：應激對照組的肝臟T-AOC顯著下降,脂質過氧化物MDA水平顯著升高(P0.05)。而添加維生素C、維生素E、槲皮素和止痢草油均能提高T-AOC,降低MDA水平(P0.05)。與非應激對照組相比,應激對照組均顯著降低了肝臟的SOD和GSH-Px活力(P0.05),添加槲皮素和止痢草油組可以提高SOD和GSH-Px活力(P0.05),而添加α-硫辛酸組肝臟的SOD和GSH-Px活力與應激對照組沒有顯著差異(P0.05)。 6)大鼠空腸組織結構形態(tài)分析：應激對照組顯著提高了隱窩深度,顯著降低了空腸絨毛高度和絨毛高度與隱窩深度比(P0.05)。添加維生素E、a-硫辛酸、槲皮素和止痢草油均能顯著提高絨毛高度(P0.05)；添加維生素C、維生素E、槲皮素均能降低隱窩深度(P0.05)；添加這5種抗氧化劑均能顯著提高絨毛高度與隱窩深度比(P0.05)。各日糧處理組的空腸絨毛高度、隱窩深度、絨毛高度與隱窩深度比,與非應激對照組相比差異不顯著(P0.05)。 7)大鼠空腸HSps mRNA表達量的變化：應激對照組與非應激對照組相比,大鼠空腸HSp27、Hsp70、Hsp90mRNA表達量均顯著升高(P0.05)；日糧添加維生素C、α-硫辛酸、維生素E、槲皮素和止痢草油均能顯著降低大鼠空腸Hsp27、Hsp70、 Hsp90mRNA表達量(P0.05)。 綜上所述,本研究的結論是：1)由體外試驗可得,槲皮素和維生素E清除DPPH自由基和抑制脂質自氧化的效果較好。2)日糧中添加維生素E、槲皮素和止痢草油均能增加動物采食量和體重,改善動物的生長性能。3)由大鼠運輸應激模型可知,補充抗氧化劑均能降低大鼠血清和肝臟中脂質過氧化,提高抗氧化酶SOD和GSH-Px活力,提高機體抗氧化能力。其中α-硫辛酸在動物體內發(fā)揮自由基清除能力、抑制脂質自氧化的作用最強,止痢草油對機體總抗氧化能力增強作用較明顯,槲皮素、維生素C、維生素E在抑制脂質自氧化和增強抗氧化酶活力均有效果,這5種抗氧化劑在運輸應激大鼠中均具有一定的抗運輸應激作用。綜合指標的測定可知,維生素E、槲皮素和止痢草油在運輸應激大鼠中抗運輸應激效果較好。
[Abstract]:The transport of animals is an important link in animal production. Transportation stress has a series of negative effects on animal production and health. The oxidative stress in the body is caused by the imbalance of oxidation and antioxidation in the body after transportation stress. The aim of this study is to evaluate the antioxidant vitamin C, vitamin E, alpha lipoic acid and mistletoe. The antioxidative ability and characteristics of PEP and SCR were compared, and the effect of these 5 antioxidants on transport stress was compared by establishing a rat model of transport stress, which provided a theoretical basis for the nutrition regulation of livestock stress. The main contents and results were as follows:
1, evaluation and characterization of antioxidant effects of different antioxidants in vitro. Using DPPH to determine the ability of vitamin C, vitamin E, quercetin and dysentery oil to remove DPPH free radicals, the results showed that the antioxidants removed the DPPH free radical IC50 as follows: vitamin C 0.0059mg/ml, vitamine E 0.0249mg/ml, quercetin as 0.0051mg/ml, dysentery The order of the 4 antioxidants is 0.0687mg/ml, the order of the scavenging ability of these 4 antioxidants to DPPH radicals is quercetin vitamin E, vitamin E, antidysentery oil. The lipid self oxidation inhibition rate of vitamin C, a- lipoic acid, vitamin E, quercetin and antidysentery oil is determined by establishing rat liver homogenate lipid autoxidation model, and the IC50 of each antioxidant is as follows. Vitamin C is 0.255mg/ml, vitamin E is 0.0052mg/ml, alpha lipoic acid is 2.06mg/ml, quercetin is 0.0058mg/ml, and dysentery oil is 0.067mg/ml, and the order of inhibiting lipid self oxidation is: vitamin E quercetin vitamin C alpha - lipoic acid.
2, the anti stress effect of antioxidant on stress rats was studied. 70 male Wistar rats of SPF grade 7-8 weeks old were randomly divided into 7 groups, with 10 repetitions in each group and 1 rats each. The non stress control group and the stress control group were fed with basal diet, and 5 diet treated groups added 1.5g/kg to the basal diet. C, 2g/kg alpha lipoic acid, 1g/kg vitamin E, 1g/kg quercetin, 100mg/kg dysentery oil. Test period 28d. After the test period, the stress group rats were treated according to the simulation conditions of transport stress (rats were placed in the constant temperature rocking bed at 35 degrees C, 60r/min, 2h). After the transport of stress treatment, the rats were anaesthetized and slaughtered to collect samples and determine serum hormones. Levels, serum and liver antioxidant index, jejunal morphology and jejunal mRNA expression.
1) adding dysentery oil could significantly increase the weight of rats, daily gain and daily intake (P0.05), and vitamin E significantly increased the weight of rats (P0.05), and the addition of quercetin could significantly increase the average daily feed intake (P0.05) of rats.
2) transport resulted in a significant decrease in body weight and a significant increase in body temperature (P0.05). There was no significant difference between weight loss and stress control group (P0.05) after the addition of all antioxidants (P0.05). The body temperature of rats in group C after transportation was significantly higher than that of other groups (P0.05).
3) the levels of cortisol and norepinephrine in the serum of stress control group were significantly increased after transportation stress treatment (P0.05). Compared with the stress control group, dietary vitamin E significantly decreased norepinephrine level (P0.05).
4) compared with the non stress group, the serum T-AOC of the stress control group was significantly lower (P0.05), the level of MDA increased significantly (P0.05), and the activity of SOD and GSH-Px decreased significantly (P0.05). After the addition of antioxidants, the level of MDA decreased significantly (P0.05), SOD activity increased significantly (P0.05), vitamin C, vitamins and dysentery. T-AOC increased significantly in grass oil group, while GSH-Px activity in vitamin E, quercetin, a- lipoic acid and Zhili grass oil group increased significantly (P0.05).
5) the effect of antioxidant on the antioxidant function of liver in stress rats: the liver T-AOC in stress control group decreased significantly and the level of lipid peroxide MDA increased significantly (P0.05). The addition of vitamin C, vitamin E, quercetin and antidysentery oil could increase T-AOC and decrease MDA level (P0.05). Compared with non stress control group, the stress control group was all compared with the control group. The activity of SOD and GSH-Px in the liver (P0.05) was significantly reduced. The activity of SOD and GSH-Px (P0.05) was increased by the addition of quercetin and antidysentery oil, while the activity of SOD and GSH-Px in the liver of the group adding alpha lipoic acid was not significantly different from that of the stress control group (P0.05).
6) the morphological analysis of the jejunum tissue: the stress control group significantly increased the depth of the recess, significantly reduced the height of the jejunum and the height of the villi and the depth of the recess (P0.05). The addition of vitamin E, a- lipoic acid, quercetin and antidysentery oil significantly increased the height of the villi (P0.05), and vitamin C, vitamin E and quercetin were all reduced. Low recess depth (P0.05); adding these 5 antioxidants could significantly increase the ratio of the height of the villi to the depth of the recess (P0.05). The height of jejunum villus, the depth of the recess, the height of the villi and the depth of the recess were not significant (P0.05).
7) the changes in the expression of HSps mRNA in the jejunum of rats: the expression of HSp27, Hsp70 and Hsp90mRNA in the jejunum of the rats increased significantly (P0.05) in the stress control group compared with the non stress control group. The diet supplemented with vitamin C, alpha lipoic acid, vitamin E, quercetin and antidysentery oil significantly reduced the expression of Hsp70 and Hsp90mRNA (P0.05) in the rat jejunum.
To sum up, the conclusions of this study are as follows: 1) in vitro tests can be obtained, quercetin and vitamin E can remove DPPH free radicals and inhibit lipid autoxidation, the effect of vitamin E, quercetin and antidysentery oil can increase animal food intake and weight, improve animal growth performance.3) by rat model of transport stress, supplemented by rat transport stress model. Antioxidants can reduce lipid peroxidation in serum and liver of rats, increase the activity of antioxidant enzyme SOD and GSH-Px, and improve the antioxidant capacity of the body. Among them, alpha lipoic acid plays the free radical scavenging ability in animals and inhibits lipid self oxidation. Vitamin C and vitamin E have effect on inhibiting lipid autoxidation and enhancing antioxidant enzyme activity. These 5 kinds of antioxidants have a certain anti transport stress in transport stress rats. The comprehensive index shows that vitamin E, quercetin and antidysentery oil have better effect on transportation stress in the transport stress rats.
【學位授予單位】：華中農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：S852.2

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