本文關(guān)鍵詞： 止痢草油 運(yùn)輸應(yīng)激 氧化應(yīng)激 肌肉狀態(tài) 肉質(zhì)　出處：《華中農(nóng)業(yè)大學(xué)》2016年博士論文　論文類(lèi)型：學(xué)位論文

【摘要】：養(yǎng)豬生產(chǎn)中運(yùn)輸應(yīng)激的發(fā)生,給豬肉屠宰、零售及相關(guān)產(chǎn)業(yè)造成巨大的經(jīng)濟(jì)損失。運(yùn)輸應(yīng)激發(fā)生時(shí),動(dòng)物應(yīng)激激素指標(biāo),各組織器官生理生化指標(biāo)、抗氧化指標(biāo)和腸道功能均發(fā)生顯著改變,肉質(zhì)的pH值、滴水損失和肉色等指標(biāo)也發(fā)生相應(yīng)變化。因此,我們推測(cè),在運(yùn)輸應(yīng)激過(guò)程中,上述指標(biāo)和功能的變化存在必然的聯(lián)系,并對(duì)肉質(zhì)造成影響。植物提取物止痢草油具有抗氧化、抗菌和改善腸道功能等多種特性,在動(dòng)物腸道及體內(nèi)均會(huì)發(fā)生作用,并影響機(jī)體生理代謝功能。此外,止痢草油(Oregano essential oils,OEO)還具有改善肉質(zhì),延長(zhǎng)肉品的貨架期的作用。本研究的目的是通過(guò)相應(yīng)試驗(yàn)研究探索止痢草油在運(yùn)輸應(yīng)激時(shí)改善肉質(zhì)的作用。本研究由三部分試驗(yàn)構(gòu)成。首先模擬運(yùn)輸構(gòu)建大鼠運(yùn)輸應(yīng)激模型,確認(rèn)模擬運(yùn)輸能夠影響大鼠肌肉氧化狀態(tài)和形態(tài)特征,通過(guò)對(duì)應(yīng)激激素、抗氧化酶活和肌肉形態(tài)等指標(biāo)檢測(cè),驗(yàn)證大鼠對(duì)運(yùn)輸應(yīng)激具有敏感性及重復(fù)性,明確模擬運(yùn)輸模型用于肌肉能量代謝和形態(tài)方面研究的可行性。然后,利用此模型研究止痢草油對(duì)運(yùn)輸應(yīng)激大鼠血液應(yīng)激激素指標(biāo),血液、腸道及肌肉抗氧化指標(biāo),腸道炎性因子及肌肉能量代謝等指標(biāo)的影響,揭示止痢草油緩解大鼠肌肉損傷的機(jī)制。最后,將止痢草油應(yīng)用在育肥豬生產(chǎn)中,通過(guò)在育肥豬飼料中添加止痢草油,研究止痢草油對(duì)育肥豬生長(zhǎng)性能的影響,并通過(guò)設(shè)定屠宰前不同的運(yùn)輸強(qiáng)度,研究止痢草油對(duì)宰前育肥豬血清、肌肉及肝臟抗氧化能力和肉質(zhì)相關(guān)指標(biāo)的影響,確定止痢草油對(duì)緩解運(yùn)輸應(yīng)激及改善肉質(zhì)的效果。主要結(jié)果如下：1、運(yùn)輸應(yīng)激大鼠模型用于肌肉狀態(tài)改變研究的可行性選擇6周齡健康雄性SD大鼠,模型溫度設(shè)定35℃,搖床轉(zhuǎn)速60 r/min,持續(xù)搖擺2 h。與對(duì)照組大鼠相比,模擬運(yùn)輸應(yīng)激大鼠血清中皮質(zhì)醇、去甲腎上腺素及促腎上腺皮質(zhì)激素釋放因子(Corticotropin releasing factor,CRF)水平極顯著升高(P0.01),血清中脂質(zhì)過(guò)氧化產(chǎn)物丙二醛(Malondialdehyde,MDA)含量極顯著升高(P0.01),而谷胱甘肽過(guò)氧化物酶(Glutathione peroxidase,GSH-Px)的活性極顯著降低(P0.01),過(guò)氧化氫酶(Catalase,CAT)的活性顯著降低(P0.05)。同時(shí),模擬運(yùn)輸應(yīng)激大鼠肌肉中MDA含量極顯著升高(P0.01),GSH-Px酶活性極顯著降低(P0.01),CAT的活性顯著降低(P0.05)。肌肉組織中活性氧(Reactive oxygen species,ROS)含量極顯著升高(P0.01)。從肌肉組織形態(tài)學(xué)觀察到,肌肉纖維有明顯的斷裂、破碎現(xiàn)象,說(shuō)明肌肉形態(tài)已經(jīng)發(fā)生損傷。這些結(jié)果表明：模擬運(yùn)輸應(yīng)激大鼠已發(fā)生明顯的應(yīng)激反應(yīng),此模型可以用于研究影響肌肉狀態(tài)的因素。2、止痢草油對(duì)運(yùn)輸應(yīng)激大鼠肌肉狀態(tài)的影響及機(jī)制的研究利用模擬運(yùn)輸應(yīng)激大鼠模型,采用雙因素試驗(yàn)設(shè)計(jì)。選擇6周齡健康雄性SD大鼠36只,隨機(jī)分為4組,每組9只。試驗(yàn)分組如下,非應(yīng)激組、應(yīng)激組、非應(yīng)激組+OEO(灌服5mg/t.bw)和應(yīng)激組+OEO(灌服5mg/t.bw)。試驗(yàn)期為20 d,記錄每天的采食量和試驗(yàn)?zāi)┲�。試�?yàn)結(jié)束后,采集血樣、肌肉和空腸等樣品。檢測(cè)應(yīng)激激素指標(biāo)、生理生化指標(biāo)、機(jī)體抗氧化指標(biāo)、肌肉形態(tài)學(xué)指標(biāo)和腸道免疫等指標(biāo),研究了止痢草油對(duì)大鼠肌肉狀態(tài)的影響及機(jī)制。結(jié)果表明：盡管添加止痢草油不能顯著提高大鼠日增重,但可以降低料重比(P0.05)。與非應(yīng)激組相比,運(yùn)輸應(yīng)激極顯著提高了大鼠血清中的皮質(zhì)醇、去甲腎上腺素、CRF、乳酸脫氫酶(Lactate dehydrogenase,LDH)、肌酸激酶(Creatine kinase,CK)、IL-6、TNFα、ROS、MDA和CAT的水平(P0.01)。與未灌服止痢草油組相比,灌服止痢草油可以極顯著降低血清中皮質(zhì)醇、去甲腎上腺素、CRF、LDH、IL-6、TNFα、ROS、MDA和GSH-Px等的水平(P0.01)。此外,皮質(zhì)醇、去甲腎上腺素、CRF、MDA、GSH-Px和SOD存在顯著的交互作用(運(yùn)輸×OEO, P0.05)。與應(yīng)激組相比,灌服止痢草油可以顯著提高宰后0h及48 h肌肉中SOD酶活性(P0.05)；極顯著降低0h及48 h肌肉MDA和乳酸濃度(P0.05)；極顯著升高0h及48 h肌肉糖原濃度(P0.01)。此外,灌服止痢草油可以提高顯著提高腸道SOD與GSH-Px酶活性(P0.01),顯著降低MDA含量(P0.05)。肌肉切片結(jié)果顯示,灌服止痢草油后,肌肉形態(tài)結(jié)構(gòu)更完整,解剖后內(nèi)臟組織無(wú)充血、脹氣現(xiàn)象。這些結(jié)果顯示,(1)止痢草油通過(guò)提高腸道抗氧化能力,降低炎性因子的產(chǎn)生,從而降低糖皮質(zhì)激素的水平,調(diào)節(jié)肌肉能量代謝,減少糖無(wú)氧酵解及乳酸的產(chǎn)生,防止pH值快速下降；(2)止痢草油通過(guò)提高機(jī)體抗氧化酶的活性,減少體內(nèi)自由基的產(chǎn)生；(3)止痢草油由于自身具有抗氧化能力,在體內(nèi)代謝過(guò)程中,能夠淬滅自由基,減少氧化產(chǎn)物的產(chǎn)生,從而減少對(duì)肌肉的損傷。3、止痢草油對(duì)肥育豬生產(chǎn)性能的影響及對(duì)肉質(zhì)的干預(yù)作用選擇日齡為150±3 d、體重78±5kg、公母各半的杜長(zhǎng)大商品豬288頭,隨機(jī)分成3組,每組96頭,分別飼養(yǎng)在6個(gè)欄里面(16頭/欄)進(jìn)行飼喂和管理。分別飼喂基礎(chǔ)日糧或添加200mg/kg維生素E、500mg/kg的止痢草油產(chǎn)品.(含OEO 5%)的日糧30 d。記錄初始重、末重和采食量等數(shù)據(jù)。飼喂結(jié)束后,每組挑選出60頭體重接近,共計(jì)180頭豬,根據(jù)模擬運(yùn)輸應(yīng)激條件,進(jìn)行高密度(252 kg/m2)長(zhǎng)時(shí)間(5h)或低密度(168 kg/m2)短時(shí)間運(yùn)輸(1 h)運(yùn)送至屠宰場(chǎng)進(jìn)行屠宰。采集血液和相關(guān)組織。結(jié)果顯示：與對(duì)照組相比,止痢草油組結(jié)束體重(P=0.06)和總增重(P=0.08)均有提高的趨勢(shì),能夠顯著降低料肉比(P0.05)。與非應(yīng)激組相比,運(yùn)輸應(yīng)激處理血清中皮質(zhì)醇水平(P=0.09)、去甲腎上腺素水平(P=0.06)和MDA水平(P=0.09)有增加趨勢(shì)；而OEO和VE處理,極顯著抑制了血清中皮質(zhì)醇和GSH-Px的水平(P0.01)；且皮質(zhì)醇和去甲腎上腺素存在極顯著的交互作用(P0.01)。與非應(yīng)激組相比,運(yùn)輸應(yīng)激處理肝臟中MDA水平有提高的趨勢(shì)(P=0.05),極顯著的提高了SOD和T-AOC的活性及ROS的水平(P0.01)：而止痢草油處理,則極顯著降低了MDA水平(P0.01),顯著提高了SOD的活性(P=0.01)。在肌肉組織中,運(yùn)輸應(yīng)激處理極顯著提高了MDA、SOD和GSH-Px的水平(P0.01)；且MDA水平存在顯著的交互作用(P=0.04)。運(yùn)輸應(yīng)激,極顯著降低了pH(45min) (P0.01),顯著降低了導(dǎo)電率(72 h)(P=0.04),顯著提高肌肉滴水損失(P=0.05)。日糧處理及運(yùn)輸方式對(duì)肌肉色值及肌肉導(dǎo)電率的影響差異不顯著(P0.05)；基礎(chǔ)日糧、VE和止痢草油處理組肌肉滴水損失差異顯著(P=0.05)。添加止痢草油極顯著地降低了肌肉中HSP27和HSP70mRNA的豐度(P0.01)；并且運(yùn)輸方式和日糧處理對(duì)HSP27的表達(dá)存在極顯著的交互作用(P0.01)；添加止痢草油組空腸絨毛高度、絨毛表面積、絨毛／隱窩比值顯著高于對(duì)照組(P0.05)。綜上所述,本研究的結(jié)論是：(1)建立了大鼠運(yùn)輸應(yīng)激模型,其條件為溫度為35℃,搖床轉(zhuǎn)速為60 r/min,持續(xù)搖擺時(shí)間為2 h。(2)止痢草油通過(guò)三個(gè)方面影響肌肉能量代謝和形態(tài)特征：第一,通過(guò)降低腸道氧化損傷,減少炎性因子的產(chǎn)生,從而降低糖皮質(zhì)激素的分泌,減少肌肉以無(wú)氧酵解方式供能,減少乳酸的積累,防止pH值快速下降；第二,止痢草油通過(guò)提高機(jī)體抗氧化酶的活性,使機(jī)體處于氧化還原平衡狀態(tài),減少氧化應(yīng)激產(chǎn)生的ROS對(duì)肌肉造成的損傷；第三,止痢草油在體內(nèi)代謝過(guò)程中,通過(guò)防止肌肉細(xì)胞膜發(fā)生脂質(zhì)氧化,減少M(fèi)DA的產(chǎn)生。(3)在育肥豬日糧中添加500 g/t的止痢草油產(chǎn)品,可以提高宰前30天肥育豬的生長(zhǎng)性能。可以緩解高密度(252kg/m2)、長(zhǎng)時(shí)間(5h)運(yùn)輸對(duì)豬應(yīng)激反應(yīng)。降低了應(yīng)激激素水平,提高了機(jī)體的抗氧化能力,改善了腸道的健康,有效緩解了運(yùn)輸應(yīng)激所帶來(lái)的負(fù)效應(yīng)。在肌肉組織中,減少了肌肉滴水損失,改善肉色等指標(biāo)。且止痢草油對(duì)肉質(zhì)的改善效果好于維生素E。
[Abstract]:Pig production in transport stress, to pork slaughter, resulting in huge economic losses in retail and related industries. Transportation stress occurs when the animal stress hormone index in various tissues of physiological and biochemical index, antioxidant index and intestinal function changed significantly, fleshy pH value, drip loss and color index changes. Therefore, we speculate that the stress in the transport process, the inevitable link change in the index and function, and its impact on meat quality. Plant extracts zhilishen oil with antioxidant, antibacterial and improve intestinal function and other characteristics, takes effect in the animal gut and the body will affect the body, and physiological function. In addition, zhilishen oil (Oregano essential oils, OEO) can improve meat quality, prolong the shelf life of meat. The purpose of this study is to explore the corresponding experimental research on dysentery Grass oil to improve meat quality in the transport stress effect. This study consists of three parts. The first test to simulate the transport construction of rat model of transport stress, confirm the simulation of transport can affect muscle oxidative status and morphology of rats, the stress hormone, detection of anti oxidase activity and muscle morphology indexes of rats with sensitivity test and the repetition of transport stress, clear the transport model to simulate feasibility study for the metabolic and morphological muscle energy. Then, the model of using zhilishen oil on the index, blood stress hormones in rat blood and intestinal transport stress, muscle antioxidant index, intestinal inflammatory factor and muscle energy metabolism. Reveal zhilishen oil relief mechanism of rat muscle injury. Finally, the zhilishen oil used in pig production, by adding zhilishen oil in fattening feed research, zhilishen Effect of oil on growth performance of fattening pig, and by setting different transportation intensity before slaughter, of zhilishen oil on pre slaughter fattening pig serum, muscle and liver index influencing the antioxidant ability and meat quality, determine zhilishen oil to relieve stress and improve the quality of the transportation effect. The main results are as follows: 1 the rat model of transport stress, feasibility study for the change of muscle status in 6 week old healthy male SD rats, model temperature of 35 DEG C, rotation speed of 60 r/min, 2 h. continuous swing compared with control group, serum cortisol analog transport stress rats, norepinephrine and corticotropin releasing factor (Corticotropin releasing factor, CRF) levels were significantly increased (P0.01), MDA in serum (Malondialdehyde, MDA) were significantly increased (P0.01), and glutathione peroxidase (Glut Athione peroxidase, GSH-Px) activity decreased significantly (P0.01), catalase (Catalase, CAT) activity was significantly reduced (P0.05). At the same time, the simulation of transport stress in rat muscle MDA content increased significantly (P0.01), GSH-Px enzyme activity was significantly decreased (P0.01), CAT activity was significantly reduced (P0.05). Active oxygen in muscle tissue (Reactive oxygen species, ROS) were significantly increased (P0.01). From the muscle morphology observed, muscle fibers were broken, broken phenomenon that has damaged muscle morphology. These results show that the simulation of transport stress in rats has significant stress response, this model.2 can be used to study the influencing factors of the muscle status, study the effects of stop dysentery grass oil on muscle state transport stress rats and the mechanism of the simulated transport stress rat model, using two factor experimental design. 6 week old healthy male Of 36 SD rats were randomly divided into 4 groups, 9 rats in each group. The experimental groups were as follows, the non stress group, stress group and non stress group +OEO (Ig 5mg/t.bw) and stress group (+OEO group 5mg/t.bw). The experiment lasted for 20 D, the daily feed intake and weight test at the end of the test. After collecting blood samples, muscle and jejunum samples. The detection of stress hormone indexes, physiological and biochemical indexes, antioxidation indexes, muscle morphology index and intestinal immune indexes of zhilishen oil effects on muscle state of rats and its mechanism. The results show that although adding zhilishen oil does not significantly improve the rats the weight gain, but it can reduce the feed weight ratio (P0.05). Compared with the non stress group, transport stress was significantly increased in rat serum cortisol and noradrenaline, CRF, lactate dehydrogenase (Lactate dehydrogenase, LDH), creatine kinase (Creatine kinase, CK), IL-6, TNF alpha, ROS, MDA and CAT 姘村鉤(P0.01).涓庢湭鐏屾湇姝㈢棦鑽夋補(bǔ)緇勭浉姣，

本文編號(hào)：1461394