本文關(guān)鍵詞： 山羊 丙酸 M_2受體 M_3受體 內(nèi)皮型一氧化氮合酶　出處：《揚州大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：奶牛真胃變位是奶牛業(yè)生產(chǎn)中一種常見內(nèi)科疾病,但其發(fā)病機制尚不清楚。丙酸作為反芻動物體內(nèi)揮發(fā)性脂肪酸(Volatile fatty acids, VFA)重要組成部分,有研究表明VFA蓄積對真胃運動有一定抑制作用。本實驗通過手術(shù)安裝的瘤胃瘺管向山羊瘤胃內(nèi)灌注不同劑量的丙酸,檢測丙酸對山羊血液理化指標(biāo)、真胃組織結(jié)構(gòu)及相關(guān)基因表達(dá)的影響,為真胃變位的發(fā)病機制提供實驗資料。選取健康山羊12只,體重30.0±2.6 kg,隨機分為4組：對照組、低劑量組、中劑量和高劑量組。每天向試驗組山羊瘤胃內(nèi)分別灌注0.5 g/kg、1.0 g/kg和1.2 g/kg的丙酸,對照組山羊灌注等量生理鹽水,連續(xù)灌注一周。每天灌注后4h采集血液和瘤胃液,檢測血常規(guī)、血液生化、瘤胃液pH值和VFA含量。灌注丙酸一周后,于次日手術(shù)采集山羊真胃組織樣品,用于病理組織學(xué)觀察和用熒光定量PCR法檢測M2、M3和eNOS的mRNA表達(dá)水平。試驗結(jié)果如下：1.灌注丙酸后,與對照組相比,三個試驗組山羊的肝、腎功能相關(guān)指標(biāo)和血鈣含量均未見明顯變化(P0.05),高劑量組山羊血糖含量極顯著升高(P0.01),低、中和高劑量組山羊的血液白細(xì)胞數(shù)和紅細(xì)胞數(shù)均未出現(xiàn)顯著變化(P0.05)。2.灌注丙酸后,與對照組相比,低、中和高組山羊瘤胃內(nèi)pH值均無明顯變化(P0.05)；低、中劑量組山羊瘤胃內(nèi)乙酸、丙酸和丁酸的濃度和高劑量組山羊瘤胃內(nèi)乙酸濃度均未出現(xiàn)顯著變化(P0.05)；高劑量組山羊瘤胃內(nèi)丙酸和丁酸濃度均極顯著升高(P0.01)。3.灌注丙酸后,進(jìn)行真胃壁病理組織學(xué)觀察,在低、中劑量組山羊真胃黏膜層未見明顯異常；高劑量組山羊真胃黏膜上皮形成的胃小凹消失,單層柱狀上皮細(xì)胞形態(tài)消失,固有層中腺體分布排列松散、不規(guī)則,黏膜肌層平滑肌肌纖維連續(xù)性消失。4.灌注丙酸后,在山羊真胃壁組織中見有M2、M3和eNOS的mRNA表達(dá)。與對照組相比,低、中劑量組山羊真胃黏膜層和全層組織中M2、M3和eNOS的mRNA表達(dá)量沒有顯著變化(P0.05),但高劑量組山羊真胃黏膜層和全層組織中M2和M3的mRNA表達(dá)量下降極顯著P0.01), eNOS的mRNA表達(dá)量極顯著升高(P0.01)。綜上所述,丙酸對山羊血常規(guī)和血液相關(guān)生化指標(biāo)無顯著影響,但是高劑量組山羊血糖含量升高且真胃黏膜層組織發(fā)生明顯病變,真胃黏膜層和全層組織中M2和M3的mRNA表達(dá)量下調(diào),eNOS的mRNA表達(dá)量有不同程度的上調(diào),這可能是丙酸誘導(dǎo)真胃蠕動減弱、繼而導(dǎo)致真胃變位的主要原因。
[Abstract]:Displaced abomasum in dairy cows is a common disease in a dairy cattle production, but its pathogenesis is still not clear. As ruminant animal propionic acid, volatile fatty acids (Volatile fatty, acids, VFA) is an important part of research shows that the accumulation of VFA has inhibitory effect on gastric motility. So this experiment through the installation operation of propionic acid rumen fistula to goat rumen infusion of different doses of propionic acid on detection of goat blood physicochemical index, expression of genes related to gastric tissue structure and really, to provide experimental data for the pathogenesis of displaced abomasum. Healthy mountain sheep 12, weight 30 + 2.6 kg, were randomly divided into 4 groups: control group, the low dose group, middle dose group and high dose group. Every day to test in rumen of goats were perfused with 0.5 g/kg, 1 g/kg and 1.2 g/kg propionic acid, the control group of goats perfusion saline, continuous perfusion for a week. Every day after reperfusion 4H In the blood and rumen fluid, blood routine examination, blood biochemical, rumen pH value and VFA content. One week after the surgery in the perfusion of propionic acid, collect the goat true stomach tissue samples for histological observation and real-time fluorescence quantitative PCR method was used to detect M2 expression levels of M3 and eNOS mRNA. The experimental results are as follows: 1. after perfusion of propionic acid, compared with the control group, three groups of goat liver, significant changes in renal function indexes and serum calcium were not found (P0.05), high dose group of goat blood sugar content increased significantly (P0.01), low, there were no obvious change in the number of white blood cells and red blood cells and high dose group (goat P0.05.2.) propionate, compared with the control group, low, middle and high group rumen pH value did not change significantly (P0.05); low dose group, rumen acetic acid, propionic acid and butyric acid concentrations and the high dose group of acetic acid concentration in rumen of goats were not This significant change (P0.05); high dose group rumen propionate and butyrate concentration were significantly increased (P0.01).3. perfusion after true stomach propionate, histological observation and in low dose group, the goat had no obvious abnormalities of gastric mucosa layer; gastric pits in high dose group goat gastric mucosa epithelial formation disappeared. Disappear form monolayer columnar epithelium, lamina propria glands arranged loose, irregular, muscularis mucosa and smooth muscle fiber disappeared after.4. continuous perfusion of propionic acid in gastric tissues, the goat saw M2, the expression of M3 and eNOS mRNA. Compared with the control group, low dose group, in abomasum of goats the whole layer of mucosa layer and M2, M3 and eNOS mRNA expression did not change significantly (P0.05), but M2 and M3 high dose group goat gastric mucosa layer and mRNA layer decreased significantly P0.01), eNOS mRNA expression increased significantly (P0 .01). To sum up, no significant effect of propionic acid on goat blood tests and blood biochemical indexes, but the high dose group blood sugar content increased and goat gastric mucosa tissue was really lesions, M2 and M3 in gastric mucosa tissue really layer and layer in the down-regulation of mRNA expression, eNOS expression of mRNA increased in different degrees this may be true, propionate induced gastric motility, which causes the displacement of the abomasum.

【學(xué)位授予單位】：揚州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S852.3

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