山羊及绵羊铜肠道损伤及其转运的比较研究

发布时间：2018-06-10 00:24

  本文选题：山羊 + 绵羊　； 参考：《吉林农业大学》2017年硕士论文

【摘要】：铜是动物生命活动中所必须的过渡金属元素,其还原酶参与有氧呼吸,神经递质合成及铁代谢等过程。虽然适量的铜能促进动物生长,提高免疫力增加采食量,但是铜的缺乏和过量都严重的影响着动物的健康及生产。山羊及绵羊虽然是同属不同种的动物,但其对铜的敏感性差异极大。本试验通过日粮中添加不同水平的铜,观测山羊及绵羊血液铜相关指标,不同肠段和血铜含量、肠道组织损伤相关参数影及肠道相关转运蛋白、铜伴侣因子mRNA表达量来研究铜水平对山羊及绵羊肠道的影响,探索铜在山羊及绵羊肠道铜转运过程的特点,同时从种属角度对山羊及绵羊肠道铜代谢机制提供理论基础。1.试验一:日粮铜水平对山羊(Capra hircus)肠道铜稳态平衡的影响试验采用完全随机化设计,分别将15只山羊为3组,每组5个重复,每个重复一只羊。各组分别为CCu0组(基础日粮组),CCu25组(基础日粮+25mg/kg·DM,CuSO4·5H2O),CCu100组(基础日粮+100mg/kg·DM,CuSO4·5H2O)。试验期60d,采集第60d血清和肠道各个肠段组织样品。检测血清中铜含量,铜蓝蛋白酶活,与肠道损伤相关指标和各个肠段组织铜含量,碱性磷酸酶、谷胱甘肽等与组织损伤及铜转运蛋白相关的酶活。各肠段组织形态指标,及与肠段损伤相关指标(肠道免疫因子,两种紧密连接蛋白mRNA表达量)。肠道铜转运因子mRNA表达量,及主要转运因子的蛋白表达量。结果如下:(1)血清和各肠段铜含量与日粮铜添加水平呈形同趋势,CCu100的CuZn-SOD、ALP酶活和组织铜含量显著高于其他两组(P0.05)。(2)CCu25组各肠段的VCR和两种紧密连接蛋白显著高于其他两组(P0.05),而TLR2和TNF-α的HIS显著低于其他两组(P0.05)。ALP、GSH-Px和ZnCu-SOD三种酶活性在十二指肠、空肠和结肠中与其他两组相比差异显著(P0.05)。(3)CCu25组的ATOX1和两种铜转运蛋白mRNA相对表达量显著高于其他两组(P0.05),而CCu100组的MTmRNA表达量显著高于其他两组(P0.05)。2.试验二:日粮铜水平对绵羊(Ovis aries)肠道损伤及转运的影响除试验对象为绵羊外,其设计与试验一相同。各组分为OCu0组(基础日粮组),OCu10组(基础日粮+10mg/kg·DM,CuSO4·5H20),OCu25组(基础日粮+25mg/kg·DM,Cu SO4·5H2O)。结果如下:(1)血清CP、CuZn-SOD、ALP的活性与日粮铜添加水平呈正相关,且具有显著性(P0.05),OCu25组除回肠外的铜含量显著高于其他两组(P0.05)。(2)十二指肠、回肠VCR值显著高于其他两组,而OCu25的VCR和两个铜伴侣蛋白mRNA表达显著低于其他两组(P0.05)。在OCu10组,LAP和ZnCu-SOD酶活性、TJP mRNA表达量显著高于其他两组(P0.05),而TLR2和TNF-α的蛋白表达评分(HIS)显著低于其他两组(P0.05)。十二指肠的GSH-Px酶活性显著高于其他两组(P0.05),OCu25除回肠外,三种酶活性在十二指肠、空肠具有显著性(P0.05)。OCuO组的Claudins表达量显著高于其他两组(P0.05),OCu10组的Claudins mRNA表达量在结肠显著高于其他两组(P0.05)。OCu25组的TLR2和TNF的HIS表达显著高于其他两组(P0.05)。(3)OCu10组各肠段的COX17、ATOXmRNA相对表达量显著高于其他两组(P0.05)。OCu10组的CTR1、ATP7AmRNA相对表达量在十二指肠、空肠的表达量显著高于其他两组(P0.05),OCu25组的MT表达量的显著高于OCu0组(P0.05)。3.日粮铜水平对山羊及绵羊铜肠道损伤及转运的比较研究(1)OCu25组的血清ALP酶活性显著高于CCu0组(P0.05),CCu0组的CuZn-SOD酶活性显著高于OCu0组(P0.05)。OCu25组的十二指肠、空肠、结肠的铜含量显著高于CCu25组。(2)CCu25的小肠VCR值显著高于OCu25组(P0.05)。CCu25组在十二指肠、空肠结肠的ALP、GSH-Px、ZnCu-SOD酶活性显著高于OCu25组(P0.05)。CCu25组各肠段的Claudins mRNA相对表达量显著高于OCu25组(P0.05),空肠、回肠的的TJP的mRNA相对表达量显著高于OCu25组(P0.05)。CCu25组的TLR2和TNF-α的HIS显著高于OCu25组(P0.05)。(3)CCu25组各个肠段的COX17、ATOX1、CTR1的mRNA相对表达量显著高于OCu25组(P0.05)。CCu25组的ATP7A在十二指肠、空肠、回肠的相对表达量显著高于OCu25组(P0.05)综上所述:日粮添加10mg/kg、25mg/kg·DM的CuSO4分别对绵羊、山羊肠道功能有改善作用,同时促进其肠道铜吸收和转运。日粮添加25mg/kg、100mg/kg·DM的CuSO4分别对绵羊、山羊肠道有损伤作用,不利于其肠道铜吸收和转运。
[Abstract]:Copper is a necessary transition metal element in animal life activities. Its reductase is involved in aerobic respiration, neurotransmitter synthesis and iron metabolism. Although a moderate amount of copper can promote animal growth and increase immunity and increase feed intake, the deficiency and excess of copper have seriously affected the health and production of animals. In the experiment, the copper related indexes, different intestinal segments and copper content, the related parameters of intestinal tissue damage, the intestinal related transporters and the expression of copper chaperone mRNA were used to study the copper level to the goat. And the influence of sheep intestinal tract, the characteristics of copper transport in goat and sheep intestines were explored. At the same time, the copper metabolism mechanism of goat and sheep intestine was provided by.1. test. The effect of dietary copper level on the homeostasis of copper homeostasis in the intestinal tract of goat (Capra hircus) was tested by complete randomization design, and 15 goats were used respectively. 3 groups, 5 repetitions in each group, each for one sheep. Group CCu0 (base diet group), group CCu25 (basal diet +25mg/kg / DM, CuSO4. 5H2O), CCu100 group (+100mg/kg. DM, CuSO4. 5H2O) in group CCu100 (basal diet +100mg/kg. DM, CuSO4. 5H2O). Damage related indexes and copper content in various intestinal segments, alkaline phosphatase, glutathione and other enzymes related to tissue damage and copper transporter. The morphological indexes of the intestinal segments, and the indexes related to intestinal damage (intestinal immune factors, the expression of two close connexin mRNA), the expression of intestinal copper transport factor mRNA, and the main cause of transport The results were as follows: (1) the content of copper in serum and all intestinal segments was the same as that of dietary copper. The CuZn-SOD, ALP enzyme activity and tissue copper content of CCu100 were significantly higher than those of the other two groups (P0.05). (2) the VCR and two close connexin in the CCu25 group were significantly higher than those of the other two groups (P0.05), while TLR2 and TNF- alpha HIS were significantly lower. In other two groups (P0.05).ALP, GSH-Px and ZnCu-SOD, the activity of three enzymes in the duodenum, the jejunum and the colon was significantly different from the other two groups (P0.05). (3) the ATOX1 of the CCu25 group and the relative expression of the two copper transporter mRNA were significantly higher than those of the other two groups (P0.05), while the MTmRNA expression in the CCu100 group was significantly higher than that of the other two groups (P0.05) test Two: the effect of dietary copper level on intestinal damage and transport in sheep (Ovis aries) was the same as that of sheep, the design and test were the same. Each group was divided into OCu0 group (basal diet group), group OCu10 (basal diet +10mg/kg, DM, CuSO4. 5H20), OCu25 group (+25mg/kg DM, Cu). The activity was positively correlated with the level of dietary copper (P0.05), and the copper content in the OCu25 group except the ileum was significantly higher than the other two groups (P0.05). (2) the duodenum, the ileum VCR value was significantly higher than the other two groups, while OCu25 VCR and two copper chaperone mRNA were significantly lower than the other two groups (P0.05). In OCu10 group, LAP and ZnCu-SOD enzyme activity. The expression of TJP mRNA was significantly higher than that of the other two groups (P0.05), while the protein expression score of TLR2 and TNF- alpha (HIS) was significantly lower than that of the other two groups (P0.05). The activity of GSH-Px enzyme in the duodenum was significantly higher than that of the other two groups (P0.05). Except for the ileum, OCu25, the activity of three enzymes was in the twelve finger intestine and the jejunum was significant (P0.05) in the.OCuO group. Higher than the other two groups (P0.05), the expression of Claudins mRNA in the OCu10 group was significantly higher than that in the other two groups (P0.05).OCu25 group, and the HIS expression of TLR2 and TNF was significantly higher than that of the other two groups (P0.05). (3) the relative expression of the two groups was significantly higher than that of the two groups. The expression of two finger intestine and jejunum was significantly higher than that of the other two groups (P0.05), and the expression of MT in group OCu25 was significantly higher than that in group OCu0 (P0.05).3. diet to the damage and transport of copper intestinal tract in goats and sheep (1) the serum ALP enzyme activity in the OCu25 group was significantly higher than that in the CCu0 group (P0.05), and the CuZn-SOD enzyme activity in CCu0 group was significantly higher than that of the group. The copper content in the duodenum, jejunum and colon in the OCu25 group was significantly higher than that in the CCu25 group. (2) the VCR value of the small intestine in CCu25 was significantly higher than that in the OCu25 group (P0.05).CCu25 group in the duodenum, the ALP, GSH-Px, ZnCu-SOD enzyme activity of the jejunum colon was significantly higher than that of the OCu25 group (P0.05). The relative expression of mRNA in the ileum was significantly higher than that of the OCu25 group (P0.05).CCu25 group TLR2 and TNF- a HIS significantly higher than that of the OCu25 group (P0.05). (3) the relative expression of all intestinal segments in the CCu25 group was significantly higher than that in the duodenum, jejunum and ileum. P0.05) to sum up, the dietary supplementation of 10mg/kg, 25mg/kg / DM CuSO4, respectively, can improve the intestinal function of sheep and goats, and promote the absorption and transport of copper in the intestines. The dietary 25mg/kg, 100mg/kg / DM CuSO4, respectively, have a damaging effect on the intestines of sheep and goat, which are unfavorable to the absorption and transport of copper in the intestine.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S826.5;S827.5

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