丙酸对山羊血液理化指标、真胃组织结构及相关基因表达的影响

发布时间：2018-03-01 05:03

本文关键词： 山羊丙酸 M_2受体 M_3受体内皮型一氧化氮合酶　出处：《扬州大学》2016年硕士论文　论文类型：学位论文

【摘要】：奶牛真胃变位是奶牛业生产中一种常见内科疾病,但其发病机制尚不清楚。丙酸作为反刍动物体内挥发性脂肪酸(Volatile fatty acids, VFA)重要组成部分,有研究表明VFA蓄积对真胃运动有一定抑制作用。本实验通过手术安装的瘤胃瘘管向山羊瘤胃内灌注不同剂量的丙酸,检测丙酸对山羊血液理化指标、真胃组织结构及相关基因表达的影响,为真胃变位的发病机制提供实验资料。选取健康山羊12只,体重30.0±2.6 kg,随机分为4组：对照组、低剂量组、中剂量和高剂量组。每天向试验组山羊瘤胃内分别灌注0.5 g/kg、1.0 g/kg和1.2 g/kg的丙酸,对照组山羊灌注等量生理盐水,连续灌注一周。每天灌注后4h采集血液和瘤胃液,检测血常规、血液生化、瘤胃液pH值和VFA含量。灌注丙酸一周后,于次日手术采集山羊真胃组织样品,用于病理组织学观察和用荧光定量PCR法检测M2、M3和eNOS的mRNA表达水平。试验结果如下：1.灌注丙酸后,与对照组相比,三个试验组山羊的肝、肾功能相关指标和血钙含量均未见明显变化(P0.05),高剂量组山羊血糖含量极显著升高(P0.01),低、中和高剂量组山羊的血液白细胞数和红细胞数均未出现显著变化(P0.05)。2.灌注丙酸后,与对照组相比,低、中和高组山羊瘤胃内pH值均无明显变化(P0.05)；低、中剂量组山羊瘤胃内乙酸、丙酸和丁酸的浓度和高剂量组山羊瘤胃内乙酸浓度均未出现显著变化(P0.05)；高剂量组山羊瘤胃内丙酸和丁酸浓度均极显著升高(P0.01)。3.灌注丙酸后,进行真胃壁病理组织学观察,在低、中剂量组山羊真胃黏膜层未见明显异常；高剂量组山羊真胃黏膜上皮形成的胃小凹消失,单层柱状上皮细胞形态消失,固有层中腺体分布排列松散、不规则,黏膜肌层平滑肌肌纤维连续性消失。4.灌注丙酸后,在山羊真胃壁组织中见有M2、M3和eNOS的mRNA表达。与对照组相比,低、中剂量组山羊真胃黏膜层和全层组织中M2、M3和eNOS的mRNA表达量没有显著变化(P0.05),但高剂量组山羊真胃黏膜层和全层组织中M2和M3的mRNA表达量下降极显著P0.01), eNOS的mRNA表达量极显著升高(P0.01)。综上所述,丙酸对山羊血常规和血液相关生化指标无显著影响,但是高剂量组山羊血糖含量升高且真胃黏膜层组织发生明显病变,真胃黏膜层和全层组织中M2和M3的mRNA表达量下调,eNOS的mRNA表达量有不同程度的上调,这可能是丙酸诱导真胃蠕动减弱、继而导致真胃变位的主要原因。
[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.

【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S852.3

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