半胱胺盐酸盐和两种必需氨基酸对山羊细胞免疫功能的影响
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摘要
为了研究半胱胺、蛋氨酸和赖氨酸对山羊细胞免疫功能的影响，我们进行了如下试验：
一、日粮营养和口服半胱胺盐酸盐对山羊手术前后外周血淋巴细胞转化率、血清白介素(IL-2)和皮质醇(cortisol)水平的影响
8头本地成年阉公山羊，随机分成两组：对照组和试验组。两组山羊自由采食粗料(花生秆)和饮水。整个试验分为4个期：1)术前粗料期；2)术前精料期；3)瘤胃手术期；4)十二指肠手术期。后三期两组山羊粗料日粮中均添加精料(50 g／goat·d)，此外，试验组在精料中拌入包被半胱胺盐酸盐CT2000(45mg／kg Bw·d，含半胱胺30％)；分别于术前粗料期第3天、精料期第11天，瘤胃瘘管手术后第8天，十二指肠瘘管手术后第4天和第13天，采血做淋巴细胞转化试验，并测血清中IL-2和cortisol水平。结果显示，正常生理状况下，增加日粮营养显著提高山羊淋巴细胞转化率(P＜0.05)，但未见其对血清IL-2和cortisol的作用。对照组在手术后淋巴细胞转化率和IL-2水平下降，而cortisol水平升高，但试验组淋巴细胞转化率下降的幅度低于对照组，且十二指肠手术后第4天显著高于对照组，血清IL-2和cortisol水平未见变化。十二指肠手术后第4天血清IL-2水平显著高于对照组(P＜0.05)，cortisol水平低于对照组(P=0.11)。研究表明，提高日粮营养水平可增强山羊机体淋巴细胞转化能力；而半胱胺盐酸盐可以阻止由手术造成的淋巴细胞转化能力和血清IL-2水平下降、血清cortisol水平上升，从而缓解了手术对细胞免疫功能的抑制。
二、粗料日粮时瘤胃灌注半胱胺盐酸盐和十二指肠灌注蛋氨酸和赖氨酸对山羊外周血淋巴细胞转化率、血清IL-2和cortisol水平的影响
5头本地阉公山羊，试验前安装永久性瘤胃瘘管和十二指肠瘘管。采用前后对比、自身对照的方法，整个试验分为四个期，每期14天：1)对照期：经十二指肠瘘管灌注5mL／time生理盐水，每天两次；2)蛋氨酸+赖氨酸期：将蛋氨酸和赖氨酸(以基础日粮中的干物质为准，蛋氨酸的灌喂量为干物质的0.03％，赖氨酸为0.05％)每天分两次溶解于5mL生理盐水中，从十二指肠瘘管中灌注；3)半胱胺盐酸盐期：从瘤胃瘘管中投放包被半胱胺盐酸盐CT2000(50mg／kgBW·d)，并同时经十二指肠瘘管灌注生理盐水5 mL／time，每天两次进行；4)蛋氨酸+赖氨酸+半胱胺盐酸盐期：每天两
半肤胺盐酸盐和两种必需氨基酸对山羊细胞免疫功能的影响
次从瘤胃痊管中投入半脱胺盐酸盐，同时从十二指肠痪管中灌注两种氨基酸溶解液，
剂量同前两期。分别在每期的第n天采血进行淋巴细胞转化试验，并测定血清IL一2
和cortisol水平。结果显示，添加半脱胺盐酸盐或/和蛋氨酸+赖氛酸期的淋巴细胞转
化率均显著高于对照期(P<0.05)，且血清。。rtisol水平显著低于对照期(P<0.05)o
半脱胺盐酸盐和两种必需氛基酸联合处理时IL一2水平比对照期有所升高(P司.09)，
而半脱胺盐酸盐和两种必需氨基酸单独处理时则没有变化。提示半脱胺盐酸盐和两种
必需氛基酸共同处理提高山羊的细胞免疫功能效果更优。
三、粗料添加部分精料日粮时口服半耽胺盐酸盐并十二指肠痊管灌注蛋氮酸和赖氨酸
对山羊外周血淋巴细胞转化率、血清IL一2和cortisol水平的影响
功头本地阉公山羊，试验前全部安装永久性瘤胃痊管和十二指肠痊管，并给予
30天恢复期。采用完全随机设计分成两组，对照组和试验组。两组山羊粗料日根中均
添加精料(50克/头)，此外，试验组在精料中拌入包被半脱胺盐酸盐CT2000(50mg/kg
BW.d，含半脱胺30%);同时，将蛋氨酸和赖氛酸(以基础日粮中的干物质为准，蛋
氛酸的灌喂量为干物质的0.03%，赖氨酸为0.05%)每天分两次溶解于smL生理盐水
中，从十二指肠痊管中灌注。对照组经十二指肠痊管每次灌注smL生理盐水。两组山
羊自由采食粗料(花生秆)和饮水。饲喂14天后，采血进行淋巴细胞转化试验，并测
定血清IL一2和co r t 15。1水平。结果表明，试验组的淋巴细胞转化率和血清IL一2水
平显著高于对照组(P<0.05)，但对cortisol水平没有明显影响。表明半脱胺盐酸盐
和蛋氨酸、赖氨酸联合处理能显著提高山羊的细胞免疫水平。
四、口服半耽胺盐酸盐对山羊产生新城疫弱毒苗诱导的杭体的影响
8头本地阉公山羊，分成两组:对照组和试验组。两组山羊粗料日粮中均添加精
料(5 09/goat)，试验组在精料中拌入包被半胧胺盐酸盐CT2000(45mg/kg BW·d，
含半脱胺30%)。两组山羊自由采食粗料(花生秆)和饮水。在试验开始前(零周w的
和第二周(wZ)，皮下注射新城疫IV系疫苗(4mL/body)。连续饲喂8周，每周采集
血清，进行血凝抑制试验(Hl)。结果表明，本试验条件下，对照组和试验组的山羊
血清的HI效价没有显著差异，未见半脱胺盐酸盐对山羊体液免疫的影响。
五、口服半耽胺盐酸盐对山羊脾脏指数的影响
试验四结束后，称山羊体重，宰杀山羊，取脾脏，并称其重;计算脾脏指数:脾
脏指数(spleen index)=脾脏重量/山羊体重;结果表明，试验组的脾脏指数显著
高于对照组(P< 0.01)。说明半脱胺盐酸盐对山羊的脾脏生长有显著的促进作用。
To gain insights into effects of cysteamine , methionine and lysine on celluar immunity in goats fed with different diets ,the following experiments were carried out.
1.Effect of dietary nutrition and cysteamine pre- and post- surgeries on transformation of peripheral blood lymphocyte concentrations of serum IL-2 and cortisol in goats
8 adult castrated goats fed with peanut stems, were randomly divided into 2 groups :control and experiment groups. Water was avalible at all time.They experienced 4 periods:1) Period of no supplemental dietary nutrition before surgery;2) Period of supplemental dietary nutrition before surgery;3)Period of surgery on rumen ;4) Period of surgery on duodenum. Supplemental nutrition ( 50 g /goat-d) was added to the diet of all goats; In addition ,cysteamine (45mg/kg BW-d) added only to the diet of experiment group in the last three periods.Two peripheral blood samples were collected at different times(the 3rd day of period 1, 11th day of period 2,8th day of period 3 ,4th and 13th days of period 4) respectively). One was used for test of transformation of peripheral blood lymphocyte(TLC) in 30 min,and the other was separated for serum and stored at -20 C until RIA assay of concentrations of IL-2 and cortisol. Results showed supplemental nutrition exerted significant effect on TLC( P<0.05 ) in normal physiological
condition,but no effect on level of serum IL-2 and cortisol. Surgery induced the decreases of TLC and serum IL-2 ,increase serum cortisol of the control goats,but the extent of decreased TLC of experiment goats was lower than that of control;There were no changes of TLC and serum IL-2 after surgery in experiment group. We concluded the following results :1) the supplemental nutrition can improve the function of lymphocyte in goats; 2) surgery can induce the immunosuppress;3)cysteamine can dimish the celluar immunosuppress by preventing the decrease of TLC , concentration of serum IL-2 and increase of serum cortisol induced by surgery.
2.Effect of cysteamine administrated by ruminal fistula or/and two kinds of essential amino acids by duodenal fistula on transformation of peripheral blood lymphocyte concentrations of serum IL-2 and cortisol in goats
5 castrated adult goats fixed permanent ruminal and duodenal fistulae a month ago experienced four periods : 1) Control period:5mL saline was administrated twice per day through duodenal fistula; 2) methionine plus lysine period: methionine(0.03%dry matter ) and lysine(0.05%dry matter) , which were dissolved in 10mL saline ervery time, were administrated twice per day through duodenal fistula ; 3)cysteamine periodxysteamine (50mg/kg BW-d ) was administrated twice per day by ruminal fistula and lOmL saline by duodenal fistula at the same time ; 4)cysteamine plus methionine plus lysine period : cysteamine was administrated by ruminal fistula and two kinds of essential amino acids by duodenal fistula at the same time with two times per day;Each period lasted 14 days.They ate peanut stems and drank water ad litum. Two peripheral blood samples were collected on the llth day of each period. One was used for test of transformation of peripheral blood lymphocyte(TLC),and the other was separated for serum and stored at -20 C until RIA assay for IL-2 and cortisol. The results showed TCL of cysteamine or/and amino acid period was significantly higher ( P<0.05 ) than that of control period ,and cortisol concentration of the last periods was lower significantly (P<0.05 ) than that of control period . IL-2 concentration of experiment period was only higher(P=0.09) than that of control period under the condition of combination of cysteamine and two essential amino acids. These results indicate the effect of the combination of cysteamine and two essential amino acids on celluar immunity is more powerful than that of either of them .
3. Effect of the combination of cysteamine orally and two amino acids administrated by duodenal fistula on transformation of peripheral blood lymphocyte concentrations of serum IL-2 and cortisol in goats
10 castrate
一、日粮营养和口服半胱胺盐酸盐对山羊手术前后外周血淋巴细胞转化率、血清白介素(IL-2)和皮质醇(cortisol)水平的影响
8头本地成年阉公山羊，随机分成两组：对照组和试验组。两组山羊自由采食粗料(花生秆)和饮水。整个试验分为4个期：1)术前粗料期；2)术前精料期；3)瘤胃手术期；4)十二指肠手术期。后三期两组山羊粗料日粮中均添加精料(50 g／goat·d)，此外，试验组在精料中拌入包被半胱胺盐酸盐CT2000(45mg／kg Bw·d，含半胱胺30％)；分别于术前粗料期第3天、精料期第11天，瘤胃瘘管手术后第8天，十二指肠瘘管手术后第4天和第13天，采血做淋巴细胞转化试验，并测血清中IL-2和cortisol水平。结果显示，正常生理状况下，增加日粮营养显著提高山羊淋巴细胞转化率(P＜0.05)，但未见其对血清IL-2和cortisol的作用。对照组在手术后淋巴细胞转化率和IL-2水平下降，而cortisol水平升高，但试验组淋巴细胞转化率下降的幅度低于对照组，且十二指肠手术后第4天显著高于对照组，血清IL-2和cortisol水平未见变化。十二指肠手术后第4天血清IL-2水平显著高于对照组(P＜0.05)，cortisol水平低于对照组(P=0.11)。研究表明，提高日粮营养水平可增强山羊机体淋巴细胞转化能力；而半胱胺盐酸盐可以阻止由手术造成的淋巴细胞转化能力和血清IL-2水平下降、血清cortisol水平上升，从而缓解了手术对细胞免疫功能的抑制。
二、粗料日粮时瘤胃灌注半胱胺盐酸盐和十二指肠灌注蛋氨酸和赖氨酸对山羊外周血淋巴细胞转化率、血清IL-2和cortisol水平的影响
5头本地阉公山羊，试验前安装永久性瘤胃瘘管和十二指肠瘘管。采用前后对比、自身对照的方法，整个试验分为四个期，每期14天：1)对照期：经十二指肠瘘管灌注5mL／time生理盐水，每天两次；2)蛋氨酸+赖氨酸期：将蛋氨酸和赖氨酸(以基础日粮中的干物质为准，蛋氨酸的灌喂量为干物质的0.03％，赖氨酸为0.05％)每天分两次溶解于5mL生理盐水中，从十二指肠瘘管中灌注；3)半胱胺盐酸盐期：从瘤胃瘘管中投放包被半胱胺盐酸盐CT2000(50mg／kgBW·d)，并同时经十二指肠瘘管灌注生理盐水5 mL／time，每天两次进行；4)蛋氨酸+赖氨酸+半胱胺盐酸盐期：每天两
半肤胺盐酸盐和两种必需氨基酸对山羊细胞免疫功能的影响
次从瘤胃痊管中投入半脱胺盐酸盐，同时从十二指肠痪管中灌注两种氨基酸溶解液，
剂量同前两期。分别在每期的第n天采血进行淋巴细胞转化试验，并测定血清IL一2
和cortisol水平。结果显示，添加半脱胺盐酸盐或/和蛋氨酸+赖氛酸期的淋巴细胞转
化率均显著高于对照期(P<0.05)，且血清。。rtisol水平显著低于对照期(P<0.05)o
半脱胺盐酸盐和两种必需氛基酸联合处理时IL一2水平比对照期有所升高(P司.09)，
而半脱胺盐酸盐和两种必需氨基酸单独处理时则没有变化。提示半脱胺盐酸盐和两种
必需氛基酸共同处理提高山羊的细胞免疫功能效果更优。
三、粗料添加部分精料日粮时口服半耽胺盐酸盐并十二指肠痊管灌注蛋氮酸和赖氨酸
对山羊外周血淋巴细胞转化率、血清IL一2和cortisol水平的影响
功头本地阉公山羊，试验前全部安装永久性瘤胃痊管和十二指肠痊管，并给予
30天恢复期。采用完全随机设计分成两组，对照组和试验组。两组山羊粗料日根中均
添加精料(50克/头)，此外，试验组在精料中拌入包被半脱胺盐酸盐CT2000(50mg/kg
BW.d，含半脱胺30%);同时，将蛋氨酸和赖氛酸(以基础日粮中的干物质为准，蛋
氛酸的灌喂量为干物质的0.03%，赖氨酸为0.05%)每天分两次溶解于smL生理盐水
中，从十二指肠痊管中灌注。对照组经十二指肠痊管每次灌注smL生理盐水。两组山
羊自由采食粗料(花生秆)和饮水。饲喂14天后，采血进行淋巴细胞转化试验，并测
定血清IL一2和co r t 15。1水平。结果表明，试验组的淋巴细胞转化率和血清IL一2水
平显著高于对照组(P<0.05)，但对cortisol水平没有明显影响。表明半脱胺盐酸盐
和蛋氨酸、赖氨酸联合处理能显著提高山羊的细胞免疫水平。
四、口服半耽胺盐酸盐对山羊产生新城疫弱毒苗诱导的杭体的影响
8头本地阉公山羊，分成两组:对照组和试验组。两组山羊粗料日粮中均添加精
料(5 09/goat)，试验组在精料中拌入包被半胧胺盐酸盐CT2000(45mg/kg BW·d，
含半脱胺30%)。两组山羊自由采食粗料(花生秆)和饮水。在试验开始前(零周w的
和第二周(wZ)，皮下注射新城疫IV系疫苗(4mL/body)。连续饲喂8周，每周采集
血清，进行血凝抑制试验(Hl)。结果表明，本试验条件下，对照组和试验组的山羊
血清的HI效价没有显著差异，未见半脱胺盐酸盐对山羊体液免疫的影响。
五、口服半耽胺盐酸盐对山羊脾脏指数的影响
试验四结束后，称山羊体重，宰杀山羊，取脾脏，并称其重;计算脾脏指数:脾
脏指数(spleen index)=脾脏重量/山羊体重;结果表明，试验组的脾脏指数显著
高于对照组(P< 0.01)。说明半脱胺盐酸盐对山羊的脾脏生长有显著的促进作用。
To gain insights into effects of cysteamine , methionine and lysine on celluar immunity in goats fed with different diets ,the following experiments were carried out.
1.Effect of dietary nutrition and cysteamine pre- and post- surgeries on transformation of peripheral blood lymphocyte concentrations of serum IL-2 and cortisol in goats
8 adult castrated goats fed with peanut stems, were randomly divided into 2 groups :control and experiment groups. Water was avalible at all time.They experienced 4 periods:1) Period of no supplemental dietary nutrition before surgery;2) Period of supplemental dietary nutrition before surgery;3)Period of surgery on rumen ;4) Period of surgery on duodenum. Supplemental nutrition ( 50 g /goat-d) was added to the diet of all goats; In addition ,cysteamine (45mg/kg BW-d) added only to the diet of experiment group in the last three periods.Two peripheral blood samples were collected at different times(the 3rd day of period 1, 11th day of period 2,8th day of period 3 ,4th and 13th days of period 4) respectively). One was used for test of transformation of peripheral blood lymphocyte(TLC) in 30 min,and the other was separated for serum and stored at -20 C until RIA assay of concentrations of IL-2 and cortisol. Results showed supplemental nutrition exerted significant effect on TLC( P<0.05 ) in normal physiological
condition,but no effect on level of serum IL-2 and cortisol. Surgery induced the decreases of TLC and serum IL-2 ,increase serum cortisol of the control goats,but the extent of decreased TLC of experiment goats was lower than that of control;There were no changes of TLC and serum IL-2 after surgery in experiment group. We concluded the following results :1) the supplemental nutrition can improve the function of lymphocyte in goats; 2) surgery can induce the immunosuppress;3)cysteamine can dimish the celluar immunosuppress by preventing the decrease of TLC , concentration of serum IL-2 and increase of serum cortisol induced by surgery.
2.Effect of cysteamine administrated by ruminal fistula or/and two kinds of essential amino acids by duodenal fistula on transformation of peripheral blood lymphocyte concentrations of serum IL-2 and cortisol in goats
5 castrated adult goats fixed permanent ruminal and duodenal fistulae a month ago experienced four periods : 1) Control period:5mL saline was administrated twice per day through duodenal fistula; 2) methionine plus lysine period: methionine(0.03%dry matter ) and lysine(0.05%dry matter) , which were dissolved in 10mL saline ervery time, were administrated twice per day through duodenal fistula ; 3)cysteamine periodxysteamine (50mg/kg BW-d ) was administrated twice per day by ruminal fistula and lOmL saline by duodenal fistula at the same time ; 4)cysteamine plus methionine plus lysine period : cysteamine was administrated by ruminal fistula and two kinds of essential amino acids by duodenal fistula at the same time with two times per day;Each period lasted 14 days.They ate peanut stems and drank water ad litum. Two peripheral blood samples were collected on the llth day of each period. One was used for test of transformation of peripheral blood lymphocyte(TLC),and the other was separated for serum and stored at -20 C until RIA assay for IL-2 and cortisol. The results showed TCL of cysteamine or/and amino acid period was significantly higher ( P<0.05 ) than that of control period ,and cortisol concentration of the last periods was lower significantly (P<0.05 ) than that of control period . IL-2 concentration of experiment period was only higher(P=0.09) than that of control period under the condition of combination of cysteamine and two essential amino acids. These results indicate the effect of the combination of cysteamine and two essential amino acids on celluar immunity is more powerful than that of either of them .
3. Effect of the combination of cysteamine orally and two amino acids administrated by duodenal fistula on transformation of peripheral blood lymphocyte concentrations of serum IL-2 and cortisol in goats
10 castrate
引文
1．刘建欣，郑昌学．现代免疫学-免疫的细胞和分子基础[M]．清华大学出版社(第二版)，2002；
2．管敏强，易礼胜．家禽饲料营养与免疫研究[J]．饲料研究，2002，1：13～15；
3．周光炎主编．免疫学原理[M]．上海科学技术出版社(第一版)；
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