【作者】 江倩；

【导师】 刘兴友； 王艳玲；

【作者基本信息】 河南农业大学 ， 基础兽医学， 2007， 硕士

【摘要】 本试验建立了定量测定罗曼蛋鸡血液和组织中囊素三肽的方法——高效液相色谱法。利用该方法分离测定了罗曼蛋鸡组织和血液中囊素三肽含量,初步发现了其随日龄增长的消长规律。选用1日龄罗曼蛋雏鸡360只,随机分成连续采血组(30只)和宰杀测试组(330只),按常规饲养条件进行饲养。宰杀测试组从1日龄到26周龄分29次宰杀取样,连续采血组心脏采血取样。采用高速冷冻离心超滤法除杂、紫外高效液相色谱法(HPLC)检测罗曼蛋鸡血液、法氏囊、脾脏和脊髓中囊素三肽的含量。色谱条件为:流动相为V(超纯水)∶V(乙腈)∶V(三氟乙酸)=80∶20∶0.1,流速为1.0mL/min,柱温25℃,检测波长215nm,进样量10ul。在蛋鸡的血液、法氏囊、脾脏、脊髓中均能检测出囊素三肽,但其含量存在较大差异。法氏囊、血液、脾脏中囊素三肽含量随日龄的增长均呈现由低到高后又降低的变化趋势。其中:法氏囊中囊素三肽的含量由1日龄的0.2836mg/mL升至8周龄达到其峰值1.6808mg/mL,52日龄到10周龄维持在高水平,均在1.60mg/mL以上,10周龄以后开始迅速下降,到18周龄时其含量低于血液中的含量,19周龄之后下降速度逐渐减缓,23周龄后渐趋平稳,但均维持在较低水平(0.37～0.40mg/mL之间),稳定之后的含量介于1周龄与2周龄之间。前期血液中的含量比法氏囊中低,在12周龄以前的变化趋势与法氏囊中基本相似,但在12周龄之后的下降速度比法氏囊中缓慢,18周龄时其含量已经高于法氏囊中的含量,23周龄后基本稳定在0.51～0.54mg/mL之间,也高于法氏囊中在此期间的含量。脾脏中的变化趋势与血液一致,含量比血液中的低。脊髓中的变化趋势有所不同,呈现12周龄之前较低,12周龄之后均保持在高位水平,但总体水平均较低。相关性分析表明,蛋鸡在12周龄之前,法氏囊为蛋鸡血液中囊素三肽的主要产生部位,血液中的囊素三肽主要与法氏囊有关。12周龄之后法氏囊逐渐萎缩,产生囊素三肽的能力降低,血液中囊素三肽主要与脊髓有关。蛋鸡在法氏囊退化后,体内还存在其它囊素三肽源器官,脊髓可能是其中的重要器官之一。脾脏中的囊素三肽是通过血液转运而来的,脾脏不是囊素三肽源器官。本研究初步获得了蛋鸡体内囊素三肽的含量变化规律,为进一步研究囊素三肽的应用及囊素三肽与疾病感染的关系奠定了实验基础。更多还原

【Abstract】 This experiment established a method,High Performance Liquid ChromatograpHy to separate and analysis bursin from layers’ blood and organs.With this method,the rise and fall regularity of bursin content with days’ increasing has been found.The study selected one-day-old Roman chickens and randomly divided them into 2 groups,groupⅠand groupⅡ.GroupⅠcontained 330 chickens and groupⅡcontained 30 chickens.The two groups were seperately abataged group and benificed for blood group.Chickens in groupⅠwere abataged for blood,bursa,spiced and spinal cord,and there were 29 duplications test from 1st day to 26th week.At the same time,chickens in groupⅡwere benificed for blood.After high speed freezing ccntrifugalization, etc,HPLC with ultraviolet ray detection was used to detect bursin in chickens’ blood and tissues.Detections conditions of HPLC were the following:mobile phase components with volume proportion were ultrapure water:acetonitrile:TFA=80:20: 0.1,speed of mobile phase is 1.0 mL/min,temperature of the column is 25℃,the detection wavelength is 215nm,the quantity of injection is 10ul.The verification experiment indicated that the detection method established has good reproducibility and high precision,and the apparatus function is stable.Bursin was found in layers’s blood,bursa,spleen and spinal cord,but bursin content in blood and erery tissue was different.With days increasing,the values of bursin content in bursa,blood and spleen were all small firstly,then large,and little at last.In bursa,the minimum value of bursin content was 0.2836 mg/mL when the chickens were one day old,and the maximal value was 1.6808 mg/mL when the layerss were 8 weeks old.The value was more than 1.60 mg/mL from 52nd day to 10th week,then the value began to become little quickly.From 18th week on,the value was less than that of blood.After 19th week,the descent speed began to become little gradually and it began to be stable but little after 23rd week.The stable value was larger than 0.37 mg/mL but less than 0.40 mg/mL,and it was larger than that of week 1 but less than that of week 2.Early days,the value of bursin content in blood was less than that of bursa and the change tendency in blood was similar to that in bursa.The value’s descent speed was less than that of bursa after 12th week.From 18th week on,it was larger than that of bursa and it was larger than 0.51 mg/mL but less than 0.54 mg/mL after 23rd week.And it was larger than that of bursa at the same term.The change tendency of bursin content in spleen was consistent with that of blood but the value was less than that of blood.In spinal cord,the bursin content value’s change tendency was different from those of bursa and blood. Correspondingly,the value was little before 12th week and after 12th week,the values kept large.But the value was little throughout.Results of the correlation analysis showed that the change tendency of bursin in blood had close relation with that in bursa before 12th week and bursin in blood had close relation with bursa.From 12th week,the bursa began to degenerate gradually and bursin in blood had clode relation with spinal cord.We can confer that other organs that can produce bursin resist in layers’ organism after the bursa’s deneneration, and spinal cord maybe one kind of them.we also confer that bursin in spleen is from bloood and spleen itself can’t produce bursin.This study found the change tendencies of bursin content in chickens’ blood and tissues and make it the base of study to bursin application and the relation between bursin and disease infection.更多还原