硒和维生素E缓解肉鸡氧化应激的研究
摘要
本研究用地塞米松（Dexamethasone, DEX）诱导建立氧化应激肉鸡模型，通过三个试验较为系统地研究了宰前氧化应激对肉鸡肉品质的影响以及日粮中添加硒和维生素E对氧化应激的缓解作用及机制，旨在为建立肉鸡抗应激营养调控技术奠定基础。
试验一研究了宰前氧化应激对肉鸡肉品质的影响。选用24只健康且体重相近的39日龄科宝500（Cobb500）肉鸡，随机分配到3个处理中，第1处理组注射生理盐水（对照组），第2处理组注射DEX3mg/kg BW，第3处理组注射DEX6mg/kg BW，连续注射3天。结果表明：宰前氧化应激显著降低肉鸡日均采食量（P<0.05），与对照组相比，DEX注射组体重显著下降（P<0.05）；屠宰率及腿肌率降低（P<0.05），胸肌率差异不显著（P>0.05）；胸肌pH24h显著下降（P<0.05），且其a*值和失水率显著升高（P<0.05），但其剪切力值差异不显著（P>0.05）；腿肌pH45min及pH_24h均极显著下降（P<0.01），其剪切力值极显著升高（P<0.01），但其失水率差异不显著（P>0.05）；血浆、胸肌及腿肌MDA含量显著升高（P<0.05），血浆GSH-Px、T-AOC与CAT活性显著升高（P<0.05）。本试验结果显示应激状态下肉鸡机体抗氧化系统会产生抗应激保护反应，这种反应虽然对维持机体生理机能有一定意义，但同时也会导致生长性能和肉品质显著下降。
试验二研究了硒和维生素E对氧化应激肉鸡生长性能、肉品质、免疫机能和抗氧化性能的影响。选用1日龄Cobb500肉鸡200只，随机分为4个处理，每个处理设5个重复，每个重复10只鸡。处理1饲喂玉米-豆粕型基础日粮，处理2、3、4分别饲喂基础日粮添加Se0.20mg/kg、VE200mg/kg、Se0.20mg/kg+VE200mg/kg的日粮。从16日龄开始，每处理随机分为A、B两组，B组腹部皮下注射DEX3mg/kg体重，隔日注射三天（16d，18d，20d），A组注射等量生理盐水。结果表明：
（1）注射期间（16~21d），试鸡日均采食量和日均增重极显著降低（P<0.01）；后期（22~42d）DEX注射对生产性能影响不显著（P>0.05）；
（2）注射DEX对42日龄肉鸡屠宰性能影响不显著（P>0.05）；
（3）21日龄时，与对照组相比，DEX注射组pH45min和pH24h显著降低（P<0.05），硒、维生素E与DEX间的互作对pH45min影响显著（P<0.05），各组间L*、a*、b*值差异不显著（P>0.05）；注射DEX后胸肌失水率显著增加（P<0.05）。42日龄时，各组间pH值、肉色、剪切力及失水率差异均不显著（P>0.05）；
（4）注射DEX极显著降低21日龄肉鸡胸腺指数和法氏囊指数（P<0.01），维生素E和DEX间互作对21日龄法氏囊指数影响显著（P<0.05）；注射DEX对42日龄肉鸡胸腺指数、脾脏指数及法氏囊指数影响均不显著（P>0.05）；与对照组相比，注射DEX显著增高21日龄血清IgA浓度（P<0.05），VE添加组血清IgA浓度显著高于对照组（P<0.01）；注射DEX对肉鸡35d和42d新城疫抗体水平影响不显著（P>0.05），添加Se和VE显著提高肉鸡35d新城疫抗体水平（P<0.05）；
（5）21日龄时，与对照组相比，注射DEX显著增高血浆MDA含量（P<0.05），极显著提高血浆、法氏囊GSH-Px活性（P<0.01）；添加VE极显著降低胸肌与法氏囊MDA含量（P<0.01），提高血浆T-AOC水平（P<0.05）；添加Se显著提高血浆CAT活性（P<0.05），降低胸肌MDA含量（P<0.05）；42日龄时，注射DEX对血浆与胸肌MDA含量、T-AOC水平及GSH-Px活性影响均不显著（P>0.05）。
这些结果揭示DEX诱导的急性氧化应激会导致鸡生长性能、肉品质及免疫机能显著降低，而在生长早期及急性应激期间日粮中添加硒和维生素E并不能完全消除应激对肉鸡的损伤效应，但日粮中添加硒和维生素E可改善肉鸡的免疫机能和抗氧化性能，缓解DEX诱导的氧化应激的负面影响，并利于肉鸡应激损伤后的康复。
试验三研究了氧化应激对肉鸡DNA损伤与甲基化的影响及VE的调控。选用1日龄Cobb500肉鸡100只，随机分为2个处理，每个处理设5个重复，每个重复10只鸡。处理1饲喂玉米-豆粕型基础日粮，处理2饲喂基础日粮添加VE200mg/kg的日粮。从16日龄开始，每处理随机分为A、B两组，B组腹部皮下注射DEX3mg/kg体重，隔日注射三天（16d，18d，20d），A组注射等量生理盐水。结果表明：注射DEX期间试鸡生长性能极显著降低（P<0.01）；肝脏、胸肌及法氏囊8-羟基脱氧鸟苷（8-OHdG）含量显著升高（P<0.05），而DNA去甲基化酶活性显著降低（P<0.05）。添加VE组注射期间日均采食量和日均增重显著高于对照组（P<0.05），8-OHdG有低于对照组趋势，但未达显著水平（P>0.05），DNA去甲基化酶活性与对照组差异不显著（P>0.05）。揭示氧化应激显著引起肉鸡DNA损伤并增高DNA甲基化程度，这可能是应激导致肉鸡性能降低的重要分子机制之一；添加维生素E可提高生长性能，减轻DNA氧化损伤，但对DNA甲基化状态无显著影响。
Three experiments were conducted in the present study to determine the effects of pres-laughter oxidative stress on meat quality and the regulations of dietary selenium and vitaminE on oxidative stress in broiler by injection of dexamethasone (DEX) in order to provide abeneficial accumulation to establish the technology of nutrient regulation against stress inbroiler industry.
In trial one, effects of preslaughter oxidative stress on meat quality of broilers were in-vestigated. Twenty-four broiler chickens (Cobb500,39-day-old) with near-mean body weight(BW) were randomly divided into three following treatments: sham injection of saline (con-trol), daily subcutaneous (s.c.) injection of DEX (3mg/kg or6mg/kg of BW, respectively)for3days. The results showed that average daily feed intake and BW of chickens were sig-nificantly decreased in the DEX treatment (P<0.05), it was the same as the change of thedressing percentage and leg muscle rate, however, the variation of breast muscle rate was notsignificant difference(P>0.05). The DEX treatment significantly decreased the pH at24hoursafter slaughter of breast muscle (P<0.05), simultaneously, significantly increased the redness(a*) and water-loss rate of breast muscle (P<0.05), but had no significant influence on shearforce of breast muscle (P>0.05). The pH at45minutes,24hours after slaughter of thigh mus-cle were significantly dropped in the DEX treatment(P<0.01), while the shear force of thighmuscle was significantly augmented(P<0.05), the change of water-loss rate of thigh musclewas not significant difference (P>0.05).The plasma,breast muscle and thigh muscle MDAwere signigicantly enhanced in the DEX treatment (P<0.05), simultaneously,the plasma activ-ity of glutathione peroxidase(GSH-Px),total antioxidant capacity(T-AOC) levels and activityof catalase(CAT) were signigicantly increased(P<0.05). Our reaearch results suggested thatthe body anti-oxidative systerm could be activated when broilers exposed to oxidative stress.Although such response could be useful to prevent oxidative injury, the following sequenceresulted in decreasing growth performance and meat quality.
In trail two, effects of dietary selenium and vitamin E on growth performance,meat qual-ity,immunity and oxidation resistance in broiler chickens exposed to DEX-induced oxidativestress was evaluated. A total of200one-day-old broiler chicks (Cobb500) were randomly di- vided into four treatments with5replicates of10birds. The first treatment was fed acorn–soybean meal basal diet, while the residual treatments were fed the basal diet supple-mented with selenium0.2mg/kg, vitamin E200mg/kg, or selenium0.2mg/kg＋vitamin E200mg/kg, respectively. All the treatments were randomly divided into two groups (A, B) at16days of age. Subsequently, the group B was injected at abdomen s.c. with DEX (3mg/kgBW) for3times every other day, while the group A was injected with saline and served ascontrol. The results were as follows:
(1)The average daily feed intake and BW gain were significantly suppressed (P<0.01)during DEX injection (16~21days), while the DEX treatment had no significant influence ongrowth performance in22~42days (P>0.05).
(2)The DEX treatment had no significant impact on slaughter parameters of42-day-oldbroilers (P>0.05).
(3)At21days of age, the pH at45minutes,24hours after slaughter of thigh muscle weresignificantly dropped in the DEX treatment compared with control (P<0.05).At the same timethere was significant interaction of selenium,vitamin E and DEX treatments for pH at45mi-nutes after slaughter (P<0.05).However,the DEX treatment had no significant influence onlightness(L*), redness(a*) and yellowness(b*)(P>0.05).In addition,the water-loss rate ofbreast muscle was significantly enhanced in the DEX treatment (P<0.05).At42days of age,the variation of pH,color,shear force and water-loss rate of breast muscle were not significantdifference among the treatments (P>0.05).
(4)The DEX treatment signigicantly decreased the thymus index and bursa of fabriciusindex of21-day-old broilers (P<0.01).There was significant interaction of vitamin E and DEXtreatment on bursa of fabricius index of21-day-old broilers (P<0.05).In contrast,the DEXtreatment had no significant influence on immue organ index of42-day-old broilers(P>0.05).Furthermore, the vitamin E (P<0.01) and DEX (P<0.05) treatment significantlyaugmented the serum immunoglobulin A(IgA) of21-day-old broilers.However,the DEXtreatment had no significant effect on newcastle disease antibody level of35-day-old and42-day-old broilers(P>0.05).Dietary selenium and vitamin E significantly enhanced the new-castle disease antibody level of35-day-old broilers (P<0.05).
(5)At21days of age,the DEX treatment significantly increased the plasma MDA levels(P<0.05), activity of GSH-Px(P<0.01) and bursa of fabricius activity of GSH-Px(P<0.01).Dietary vitamin E significantly declined the breast and bursa of fabricius MDA le-vels (P<0.01) and enhanced plasma T-AOC levels (P<0.05).Dietary selenium significantlyincreased plasma activity of CAT and decreased breast muscle MDA levels (P<0.05).At42days of age,there was no significant influence on plasma and breast muscle MDA le- vels,T-AOC levels and activity of GSH-Px (P>0.05).
These results suggested that growth performance, meat quality and immunity were sig-nificantly declined when broilers suffered from acute oxidative stress induced by DEX.Moreover, dietary supplementation with selenium and vitamin E during early growth andacute oxidative stress stage could not completely eliminate the injuries of oxidative stress forbroilers.However, dietary supplementation with selenium and vitamin E was favorable forimproving immunity and oxidation resisitance to alleviate DEX-induced oxidative stress, alsocontributed to chickens retrieve from stress state to the normal.
In trail three, effects of oxidative stress on DNA damage, DNA methylation and themodulation by vitamin E were evaluated. A total of100one-day-old broiler chicks (Cobb500)were randomly and equally assigned to two treatments with5replicates.The birds were fedeither a basal diet or a basal diet supplemented with vitamin E (200mg/kg basal diet). All thetreatments were randomly divided into two groups (A,B) at16days of age. Subsequently, thegroup B was injected at abdomen s.c. with DEX (3mg/kg BW) for3times every other day,while the group A was injected with saline and served as control. The results indicated that thegrowth performance was significantly suppressd when birds exposed to DEX-induced oxida-tive stress (P<0.01).8-hydroxydeoxyguanosine (8-OHdG) concentration enhanced by DEXtreatment (P<0.05), while activity of DNA demethylase declined (P<0.05).Dietary vitamin Esignificantly augmented average daily feed intake and daily gain (P<0.05) and tended to atte-nuate8-OHdG concentration (P>0.05).However, the activity of DNA demethylase was notinfluenced by vitamin E supplementation (P>0.05). The results of current research suggestthat broilers had a poor performance under oxidative stress through increasing degree of DNAdamage or DNA methylation. Supplemented with vitamin E was useful to improve the growthperformance and alleviate DNA oxidative injury to some extent, whereas no significant effecton DNA methylation was observed.
试验一研究了宰前氧化应激对肉鸡肉品质的影响。选用24只健康且体重相近的39日龄科宝500（Cobb500）肉鸡，随机分配到3个处理中，第1处理组注射生理盐水（对照组），第2处理组注射DEX3mg/kg BW，第3处理组注射DEX6mg/kg BW，连续注射3天。结果表明：宰前氧化应激显著降低肉鸡日均采食量（P<0.05），与对照组相比，DEX注射组体重显著下降（P<0.05）；屠宰率及腿肌率降低（P<0.05），胸肌率差异不显著（P>0.05）；胸肌pH24h显著下降（P<0.05），且其a*值和失水率显著升高（P<0.05），但其剪切力值差异不显著（P>0.05）；腿肌pH45min及pH_24h均极显著下降（P<0.01），其剪切力值极显著升高（P<0.01），但其失水率差异不显著（P>0.05）；血浆、胸肌及腿肌MDA含量显著升高（P<0.05），血浆GSH-Px、T-AOC与CAT活性显著升高（P<0.05）。本试验结果显示应激状态下肉鸡机体抗氧化系统会产生抗应激保护反应，这种反应虽然对维持机体生理机能有一定意义，但同时也会导致生长性能和肉品质显著下降。
试验二研究了硒和维生素E对氧化应激肉鸡生长性能、肉品质、免疫机能和抗氧化性能的影响。选用1日龄Cobb500肉鸡200只，随机分为4个处理，每个处理设5个重复，每个重复10只鸡。处理1饲喂玉米-豆粕型基础日粮，处理2、3、4分别饲喂基础日粮添加Se0.20mg/kg、VE200mg/kg、Se0.20mg/kg+VE200mg/kg的日粮。从16日龄开始，每处理随机分为A、B两组，B组腹部皮下注射DEX3mg/kg体重，隔日注射三天（16d，18d，20d），A组注射等量生理盐水。结果表明：
（1）注射期间（16~21d），试鸡日均采食量和日均增重极显著降低（P<0.01）；后期（22~42d）DEX注射对生产性能影响不显著（P>0.05）；
（2）注射DEX对42日龄肉鸡屠宰性能影响不显著（P>0.05）；
（3）21日龄时，与对照组相比，DEX注射组pH45min和pH24h显著降低（P<0.05），硒、维生素E与DEX间的互作对pH45min影响显著（P<0.05），各组间L*、a*、b*值差异不显著（P>0.05）；注射DEX后胸肌失水率显著增加（P<0.05）。42日龄时，各组间pH值、肉色、剪切力及失水率差异均不显著（P>0.05）；
（4）注射DEX极显著降低21日龄肉鸡胸腺指数和法氏囊指数（P<0.01），维生素E和DEX间互作对21日龄法氏囊指数影响显著（P<0.05）；注射DEX对42日龄肉鸡胸腺指数、脾脏指数及法氏囊指数影响均不显著（P>0.05）；与对照组相比，注射DEX显著增高21日龄血清IgA浓度（P<0.05），VE添加组血清IgA浓度显著高于对照组（P<0.01）；注射DEX对肉鸡35d和42d新城疫抗体水平影响不显著（P>0.05），添加Se和VE显著提高肉鸡35d新城疫抗体水平（P<0.05）；
（5）21日龄时，与对照组相比，注射DEX显著增高血浆MDA含量（P<0.05），极显著提高血浆、法氏囊GSH-Px活性（P<0.01）；添加VE极显著降低胸肌与法氏囊MDA含量（P<0.01），提高血浆T-AOC水平（P<0.05）；添加Se显著提高血浆CAT活性（P<0.05），降低胸肌MDA含量（P<0.05）；42日龄时，注射DEX对血浆与胸肌MDA含量、T-AOC水平及GSH-Px活性影响均不显著（P>0.05）。
这些结果揭示DEX诱导的急性氧化应激会导致鸡生长性能、肉品质及免疫机能显著降低，而在生长早期及急性应激期间日粮中添加硒和维生素E并不能完全消除应激对肉鸡的损伤效应，但日粮中添加硒和维生素E可改善肉鸡的免疫机能和抗氧化性能，缓解DEX诱导的氧化应激的负面影响，并利于肉鸡应激损伤后的康复。
试验三研究了氧化应激对肉鸡DNA损伤与甲基化的影响及VE的调控。选用1日龄Cobb500肉鸡100只，随机分为2个处理，每个处理设5个重复，每个重复10只鸡。处理1饲喂玉米-豆粕型基础日粮，处理2饲喂基础日粮添加VE200mg/kg的日粮。从16日龄开始，每处理随机分为A、B两组，B组腹部皮下注射DEX3mg/kg体重，隔日注射三天（16d，18d，20d），A组注射等量生理盐水。结果表明：注射DEX期间试鸡生长性能极显著降低（P<0.01）；肝脏、胸肌及法氏囊8-羟基脱氧鸟苷（8-OHdG）含量显著升高（P<0.05），而DNA去甲基化酶活性显著降低（P<0.05）。添加VE组注射期间日均采食量和日均增重显著高于对照组（P<0.05），8-OHdG有低于对照组趋势，但未达显著水平（P>0.05），DNA去甲基化酶活性与对照组差异不显著（P>0.05）。揭示氧化应激显著引起肉鸡DNA损伤并增高DNA甲基化程度，这可能是应激导致肉鸡性能降低的重要分子机制之一；添加维生素E可提高生长性能，减轻DNA氧化损伤，但对DNA甲基化状态无显著影响。
Three experiments were conducted in the present study to determine the effects of pres-laughter oxidative stress on meat quality and the regulations of dietary selenium and vitaminE on oxidative stress in broiler by injection of dexamethasone (DEX) in order to provide abeneficial accumulation to establish the technology of nutrient regulation against stress inbroiler industry.
In trial one, effects of preslaughter oxidative stress on meat quality of broilers were in-vestigated. Twenty-four broiler chickens (Cobb500,39-day-old) with near-mean body weight(BW) were randomly divided into three following treatments: sham injection of saline (con-trol), daily subcutaneous (s.c.) injection of DEX (3mg/kg or6mg/kg of BW, respectively)for3days. The results showed that average daily feed intake and BW of chickens were sig-nificantly decreased in the DEX treatment (P<0.05), it was the same as the change of thedressing percentage and leg muscle rate, however, the variation of breast muscle rate was notsignificant difference(P>0.05). The DEX treatment significantly decreased the pH at24hoursafter slaughter of breast muscle (P<0.05), simultaneously, significantly increased the redness(a*) and water-loss rate of breast muscle (P<0.05), but had no significant influence on shearforce of breast muscle (P>0.05). The pH at45minutes,24hours after slaughter of thigh mus-cle were significantly dropped in the DEX treatment(P<0.01), while the shear force of thighmuscle was significantly augmented(P<0.05), the change of water-loss rate of thigh musclewas not significant difference (P>0.05).The plasma,breast muscle and thigh muscle MDAwere signigicantly enhanced in the DEX treatment (P<0.05), simultaneously,the plasma activ-ity of glutathione peroxidase(GSH-Px),total antioxidant capacity(T-AOC) levels and activityof catalase(CAT) were signigicantly increased(P<0.05). Our reaearch results suggested thatthe body anti-oxidative systerm could be activated when broilers exposed to oxidative stress.Although such response could be useful to prevent oxidative injury, the following sequenceresulted in decreasing growth performance and meat quality.
In trail two, effects of dietary selenium and vitamin E on growth performance,meat qual-ity,immunity and oxidation resistance in broiler chickens exposed to DEX-induced oxidativestress was evaluated. A total of200one-day-old broiler chicks (Cobb500) were randomly di- vided into four treatments with5replicates of10birds. The first treatment was fed acorn–soybean meal basal diet, while the residual treatments were fed the basal diet supple-mented with selenium0.2mg/kg, vitamin E200mg/kg, or selenium0.2mg/kg＋vitamin E200mg/kg, respectively. All the treatments were randomly divided into two groups (A, B) at16days of age. Subsequently, the group B was injected at abdomen s.c. with DEX (3mg/kgBW) for3times every other day, while the group A was injected with saline and served ascontrol. The results were as follows:
(1)The average daily feed intake and BW gain were significantly suppressed (P<0.01)during DEX injection (16~21days), while the DEX treatment had no significant influence ongrowth performance in22~42days (P>0.05).
(2)The DEX treatment had no significant impact on slaughter parameters of42-day-oldbroilers (P>0.05).
(3)At21days of age, the pH at45minutes,24hours after slaughter of thigh muscle weresignificantly dropped in the DEX treatment compared with control (P<0.05).At the same timethere was significant interaction of selenium,vitamin E and DEX treatments for pH at45mi-nutes after slaughter (P<0.05).However,the DEX treatment had no significant influence onlightness(L*), redness(a*) and yellowness(b*)(P>0.05).In addition,the water-loss rate ofbreast muscle was significantly enhanced in the DEX treatment (P<0.05).At42days of age,the variation of pH,color,shear force and water-loss rate of breast muscle were not significantdifference among the treatments (P>0.05).
(4)The DEX treatment signigicantly decreased the thymus index and bursa of fabriciusindex of21-day-old broilers (P<0.01).There was significant interaction of vitamin E and DEXtreatment on bursa of fabricius index of21-day-old broilers (P<0.05).In contrast,the DEXtreatment had no significant influence on immue organ index of42-day-old broilers(P>0.05).Furthermore, the vitamin E (P<0.01) and DEX (P<0.05) treatment significantlyaugmented the serum immunoglobulin A(IgA) of21-day-old broilers.However,the DEXtreatment had no significant effect on newcastle disease antibody level of35-day-old and42-day-old broilers(P>0.05).Dietary selenium and vitamin E significantly enhanced the new-castle disease antibody level of35-day-old broilers (P<0.05).
(5)At21days of age,the DEX treatment significantly increased the plasma MDA levels(P<0.05), activity of GSH-Px(P<0.01) and bursa of fabricius activity of GSH-Px(P<0.01).Dietary vitamin E significantly declined the breast and bursa of fabricius MDA le-vels (P<0.01) and enhanced plasma T-AOC levels (P<0.05).Dietary selenium significantlyincreased plasma activity of CAT and decreased breast muscle MDA levels (P<0.05).At42days of age,there was no significant influence on plasma and breast muscle MDA le- vels,T-AOC levels and activity of GSH-Px (P>0.05).
These results suggested that growth performance, meat quality and immunity were sig-nificantly declined when broilers suffered from acute oxidative stress induced by DEX.Moreover, dietary supplementation with selenium and vitamin E during early growth andacute oxidative stress stage could not completely eliminate the injuries of oxidative stress forbroilers.However, dietary supplementation with selenium and vitamin E was favorable forimproving immunity and oxidation resisitance to alleviate DEX-induced oxidative stress, alsocontributed to chickens retrieve from stress state to the normal.
In trail three, effects of oxidative stress on DNA damage, DNA methylation and themodulation by vitamin E were evaluated. A total of100one-day-old broiler chicks (Cobb500)were randomly and equally assigned to two treatments with5replicates.The birds were fedeither a basal diet or a basal diet supplemented with vitamin E (200mg/kg basal diet). All thetreatments were randomly divided into two groups (A,B) at16days of age. Subsequently, thegroup B was injected at abdomen s.c. with DEX (3mg/kg BW) for3times every other day,while the group A was injected with saline and served as control. The results indicated that thegrowth performance was significantly suppressd when birds exposed to DEX-induced oxida-tive stress (P<0.01).8-hydroxydeoxyguanosine (8-OHdG) concentration enhanced by DEXtreatment (P<0.05), while activity of DNA demethylase declined (P<0.05).Dietary vitamin Esignificantly augmented average daily feed intake and daily gain (P<0.05) and tended to atte-nuate8-OHdG concentration (P>0.05).However, the activity of DNA demethylase was notinfluenced by vitamin E supplementation (P>0.05). The results of current research suggestthat broilers had a poor performance under oxidative stress through increasing degree of DNAdamage or DNA methylation. Supplemented with vitamin E was useful to improve the growthperformance and alleviate DNA oxidative injury to some extent, whereas no significant effecton DNA methylation was observed.
引文
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