山羊及綿羊銅腸道損傷及其轉(zhuǎn)運(yùn)的比較研究
本文選題:山羊 + 綿羊 ; 參考:《吉林農(nóng)業(yè)大學(xué)》2017年碩士論文
【摘要】:銅是動(dòng)物生命活動(dòng)中所必須的過(guò)渡金屬元素,其還原酶參與有氧呼吸,神經(jīng)遞質(zhì)合成及鐵代謝等過(guò)程。雖然適量的銅能促進(jìn)動(dòng)物生長(zhǎng),提高免疫力增加采食量,但是銅的缺乏和過(guò)量都嚴(yán)重的影響著動(dòng)物的健康及生產(chǎn)。山羊及綿羊雖然是同屬不同種的動(dòng)物,但其對(duì)銅的敏感性差異極大。本試驗(yàn)通過(guò)日糧中添加不同水平的銅,觀測(cè)山羊及綿羊血液銅相關(guān)指標(biāo),不同腸段和血銅含量、腸道組織損傷相關(guān)參數(shù)影及腸道相關(guān)轉(zhuǎn)運(yùn)蛋白、銅伴侶因子mRNA表達(dá)量來(lái)研究銅水平對(duì)山羊及綿羊腸道的影響,探索銅在山羊及綿羊腸道銅轉(zhuǎn)運(yùn)過(guò)程的特點(diǎn),同時(shí)從種屬角度對(duì)山羊及綿羊腸道銅代謝機(jī)制提供理論基礎(chǔ)。1.試驗(yàn)一:日糧銅水平對(duì)山羊(Capra hircus)腸道銅穩(wěn)態(tài)平衡的影響試驗(yàn)采用完全隨機(jī)化設(shè)計(jì),分別將15只山羊?yàn)?組,每組5個(gè)重復(fù),每個(gè)重復(fù)一只羊。各組分別為CCu0組(基礎(chǔ)日糧組),CCu25組(基礎(chǔ)日糧+25mg/kg·DM,CuSO4·5H2O),CCu100組(基礎(chǔ)日糧+100mg/kg·DM,CuSO4·5H2O)。試驗(yàn)期60d,采集第60d血清和腸道各個(gè)腸段組織樣品。檢測(cè)血清中銅含量,銅藍(lán)蛋白酶活,與腸道損傷相關(guān)指標(biāo)和各個(gè)腸段組織銅含量,堿性磷酸酶、谷胱甘肽等與組織損傷及銅轉(zhuǎn)運(yùn)蛋白相關(guān)的酶活。各腸段組織形態(tài)指標(biāo),及與腸段損傷相關(guān)指標(biāo)(腸道免疫因子,兩種緊密連接蛋白mRNA表達(dá)量)。腸道銅轉(zhuǎn)運(yùn)因子mRNA表達(dá)量,及主要轉(zhuǎn)運(yùn)因子的蛋白表達(dá)量。結(jié)果如下:(1)血清和各腸段銅含量與日糧銅添加水平呈形同趨勢(shì),CCu100的CuZn-SOD、ALP酶活和組織銅含量顯著高于其他兩組(P0.05)。(2)CCu25組各腸段的VCR和兩種緊密連接蛋白顯著高于其他兩組(P0.05),而TLR2和TNF-α的HIS顯著低于其他兩組(P0.05)。ALP、GSH-Px和ZnCu-SOD三種酶活性在十二指腸、空腸和結(jié)腸中與其他兩組相比差異顯著(P0.05)。(3)CCu25組的ATOX1和兩種銅轉(zhuǎn)運(yùn)蛋白mRNA相對(duì)表達(dá)量顯著高于其他兩組(P0.05),而CCu100組的MTmRNA表達(dá)量顯著高于其他兩組(P0.05)。2.試驗(yàn)二:日糧銅水平對(duì)綿羊(Ovis aries)腸道損傷及轉(zhuǎn)運(yùn)的影響除試驗(yàn)對(duì)象為綿羊外,其設(shè)計(jì)與試驗(yàn)一相同。各組分為OCu0組(基礎(chǔ)日糧組),OCu10組(基礎(chǔ)日糧+10mg/kg·DM,CuSO4·5H20),OCu25組(基礎(chǔ)日糧+25mg/kg·DM,Cu SO4·5H2O)。結(jié)果如下:(1)血清CP、CuZn-SOD、ALP的活性與日糧銅添加水平呈正相關(guān),且具有顯著性(P0.05),OCu25組除回腸外的銅含量顯著高于其他兩組(P0.05)。(2)十二指腸、回腸VCR值顯著高于其他兩組,而OCu25的VCR和兩個(gè)銅伴侶蛋白mRNA表達(dá)顯著低于其他兩組(P0.05)。在OCu10組,LAP和ZnCu-SOD酶活性、TJP mRNA表達(dá)量顯著高于其他兩組(P0.05),而TLR2和TNF-α的蛋白表達(dá)評(píng)分(HIS)顯著低于其他兩組(P0.05)。十二指腸的GSH-Px酶活性顯著高于其他兩組(P0.05),OCu25除回腸外,三種酶活性在十二指腸、空腸具有顯著性(P0.05)。OCuO組的Claudins表達(dá)量顯著高于其他兩組(P0.05),OCu10組的Claudins mRNA表達(dá)量在結(jié)腸顯著高于其他兩組(P0.05)。OCu25組的TLR2和TNF的HIS表達(dá)顯著高于其他兩組(P0.05)。(3)OCu10組各腸段的COX17、ATOXmRNA相對(duì)表達(dá)量顯著高于其他兩組(P0.05)。OCu10組的CTR1、ATP7AmRNA相對(duì)表達(dá)量在十二指腸、空腸的表達(dá)量顯著高于其他兩組(P0.05),OCu25組的MT表達(dá)量的顯著高于OCu0組(P0.05)。3.日糧銅水平對(duì)山羊及綿羊銅腸道損傷及轉(zhuǎn)運(yùn)的比較研究(1)OCu25組的血清ALP酶活性顯著高于CCu0組(P0.05),CCu0組的CuZn-SOD酶活性顯著高于OCu0組(P0.05)。OCu25組的十二指腸、空腸、結(jié)腸的銅含量顯著高于CCu25組。(2)CCu25的小腸VCR值顯著高于OCu25組(P0.05)。CCu25組在十二指腸、空腸結(jié)腸的ALP、GSH-Px、ZnCu-SOD酶活性顯著高于OCu25組(P0.05)。CCu25組各腸段的Claudins mRNA相對(duì)表達(dá)量顯著高于OCu25組(P0.05),空腸、回腸的的TJP的mRNA相對(duì)表達(dá)量顯著高于OCu25組(P0.05)。CCu25組的TLR2和TNF-α的HIS顯著高于OCu25組(P0.05)。(3)CCu25組各個(gè)腸段的COX17、ATOX1、CTR1的mRNA相對(duì)表達(dá)量顯著高于OCu25組(P0.05)。CCu25組的ATP7A在十二指腸、空腸、回腸的相對(duì)表達(dá)量顯著高于OCu25組(P0.05)綜上所述:日糧添加10mg/kg、25mg/kg·DM的CuSO4分別對(duì)綿羊、山羊腸道功能有改善作用,同時(shí)促進(jìn)其腸道銅吸收和轉(zhuǎn)運(yùn)。日糧添加25mg/kg、100mg/kg·DM的CuSO4分別對(duì)綿羊、山羊腸道有損傷作用,不利于其腸道銅吸收和轉(zhuǎn)運(yùn)。
[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.
【學(xué)位授予單位】:吉林農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2017
【分類號(hào)】:S826.5;S827.5
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