不同营养时期牦牛组织中β-胡萝卜素含量及相关降解酶的研究

发布时间：2018-01-19 10:34

本文关键词： 牦牛 β-胡萝素维生素A β胡萝卜素-15 15　出处：《青海大学》2015年硕士论文　论文类型：学位论文

【摘要】：成年牦牛(Bos mutus)脂肪呈现出明显的黄色,其对β-胡萝卜素在内的类胡萝卜素具有较强的富集能力,这种现象并非遗传缺陷及病理原因造成的,而是牦牛适应高原特殊生境环境的一种营养代谢机制。为了研究和探讨牦牛脂肪中β-胡萝卜素富集与β-胡萝卜素关键代谢酶之间的关系,采用高效液相色谱法测定牦牛组织中β-胡萝卜素和维生素A的含量,并对β-胡萝卜素降解酶含量及酶活进行测定;应用实时荧光定量PCR检测牦牛不同营养时期组织中β胡萝卜素-15,15'-单加氧酶(BCMO1)m RNA的表达差异,应用蛋白免疫印迹(western blot)检测牦牛不同营养时期组织中β胡萝卜素-15,15'-单加氧酶(BCMO1)蛋白表达水平。(1)不同营养时期,各组织中β-胡萝卜素含量不同。营养丰盛期牦牛血清、肝脏、十二指肠、空肠和瘤胃中β-胡萝卜素的含量明显高于营养匮乏期(P0.05),其中血清中β-胡萝卜素的含量差异最明显,而脂肪和肾脏中β-胡萝卜素含量差异不显著(P0.05)。营养丰盛期肝脏中维生素A的含量明显高于营养匮乏期,两个时期相比较,回肠、肠内容物、瘤胃内容物及其他组织中维生素A含量差异显著(P0.05);(2)牦牛肝脏和胰脏组织β-胡萝卜素降解酶活性在营养丰盛期要高于营养匮乏期,而其他组织降解酶活性在营养匮乏期要高于营养丰盛期。(3)不同营养时期牦牛β胡萝卜素-15,15'-单加氧酶(BCMO1)m RNA基因在各组织中均有表达,在营养丰盛期十二指肠、肾脏、瘤胃和空肠中的m RNA的表达量明显低于营养匮乏期(P0.05),而肌肉和胰脏中m RNA的表达量营养丰盛期要高于营养匮乏期。(4)牦牛十二指肠、空肠、肾脏、肌肉和瘤胃组织中的β胡萝卜素-15,15'-单加氧酶(BCMO1)蛋白表达量,营养丰盛期均低于营养匮乏期(P0.05),肝脏中的β胡萝卜素-15,15'-单加氧酶(BCMO1)蛋白表达量在营养丰盛期要显著高于营养匮乏期(P0.05),肺脏和回肠中的β胡萝卜素-15,15'-单加氧酶(BCMO1)蛋白表达量在两个营养时期差异不显著(P0.05)。通过对研究结果的分析发现,牦牛脂肪等组织在营养丰盛期能富集大量的β-胡萝卜素,在维生素A缺乏的季节,组织中β-胡萝卜素可降解转化为VA。牦牛脂肪中β-胡萝卜素的富集与β-胡萝卜素降解酶有一定的关系,具体表现在除肝脏和胰脏外,牦牛组织中β-胡萝卜素降解酶活性在营养匮乏期高于营养丰盛期,从基因及蛋白表达水平分析在营养丰盛期大部分组织中β胡萝卜素-15,15'-单加氧酶(BCMO1)的表达水平低于营养匮乏期。从β-胡萝卜素反馈代谢机理分析β-胡萝卜素的代谢产物(视黄酸等)可能会反馈性抑制β胡萝卜素-15,15'-单加氧酶(BCMO1)的表达。
[Abstract]:The fat of adult Yak Bos Mutusus showed obvious yellow, and it had a strong ability to enrich the carotenoid, including 尾 -carotene. This phenomenon is not caused by genetic defects and pathological reasons. In order to study the relationship between 尾 -carotene enrichment and 尾 -carotene key metabolic enzymes in yak fat. The content of 尾 -carotene and vitamin A in yak tissue was determined by HPLC, and the content and activity of 尾 -carotene degrading enzyme were determined. Real time fluorescence quantitative PCR was used to detect the expression of 尾 -carotene -15A15 and monooxygenase (BCMO1) m RNA in yak tissues at different nutritional stages. Western blot was used to detect 尾 -carotene 15 in yak tissues at different nutritional stages. The expression level of BCMO1) protein was different in different nutrition period. The serum, liver and duodenum of yak during nutritious period were different. The content of 尾 -carotene in jejunum and rumen was significantly higher than that in nutrient-deficient period (P 0.05), and the difference of serum 尾 -carotene was most obvious. However, there was no significant difference in 尾 -carotene content between fat and kidney. The content of vitamin A in the liver in the nutrient-rich period was significantly higher than that in the nutrition-deficient period, and the contents of ileum and intestine were compared between the two periods. The content of vitamin A in rumen contents and other tissues was significantly different from that in other tissues. (2) the activity of 尾 -carotene degrading enzyme in liver and pancreas of yak was higher than that in the period of rich nutrition. The activity of other tissue degradation enzymes was higher in the period of nutrient deficiency than that in the period of rich nutrition.) 尾 -carotene -15 in yak at different nutrition stage. BCMO1m RNA gene was expressed in all tissues, and it was found in duodenum and kidney during nutritious period. The expression of m RNA in rumen and jejunum was significantly lower than that in nutritional deficiency period (P 0.05). The expression of m RNA in muscle and pancreas was higher than that in nutrient-deficient period. The 尾 -carotene -15 was found in duodenum, jejunum, kidney, muscle and rumen tissue of yak. The protein expression of BCMO1 was lower than that of P0.05 in nutrition-deficient period and 尾 -carotene-15 in liver. The expression of BCMO1 protein was significantly higher than that of P0.05, 尾 -carotene -15 in lung and ileum during the period of rich nutrition. There was no significant difference in the expression of BCMO1 protein between the two nutrition periods (P 0.05). It was found by the analysis of the results of the study that there was no significant difference in the expression of BCMO1 protein between the two nutrition periods. Yak fat and other tissues can enrich a large amount of 尾 -carotene during the period of rich nutrition, in the season of vitamin A deficiency. The concentration of 尾 -carotene in fat of yak was related to 尾 -carotene degrading enzyme, except liver and pancreas. The activity of 尾 -carotene degrading enzyme in yak tissue was higher than that in nutrition-rich period, and 尾 -carotene -15 in most tissues of nutrition-rich period was analyzed from the level of gene and protein expression. The expression level of BCMO1 was lower than that in the period of nutritional deficiency. The metabolites of 尾 -carotene (retinoic acid, etc.) were analyzed from the feedback metabolic mechanism of 尾 -carotene. May have a feedback inhibition of beta-carotene -15. The expression of BCMO1.
【学位授予单位】：青海大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823.85

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