纳米豆粕在小鼠消化道内示踪研究

发布时间：2019-04-19 03:23

【摘要】：本文以纳米豆粕为研究对象,主要研究内容包括纳米豆粕的制备及粒径表征;纳米豆粕在小鼠消化道各部分的分布情况的研究;纳米豆粕对小鼠消化道形态结构的影响。主要研究方法、结果与结论如下:1.采用球磨法将普通豆粕加工成纳米豆粕,通过扫描电子显微镜和激光粒度仪对纳米豆粕的形貌和粒径进行表征,结果表明:豆粕粒子粒径分布在200-300nm,单分散性良好。扫描电镜观察显示纳米豆粕粒子有的具有球形形貌特征,有的为椭圆形,还有的为不规则形状,其中球形和椭圆形所占比例较大,纳米豆粕粒子尺寸分布比较均匀。激光粒度仪分析结果显示:干磨法研磨获得的纳米豆粕最小粒径在200nm附近,粒径分布比较均匀,粒径分布峰值在219.8nm处。2.本论文利用β-伴大豆球蛋白的抗原性和相对稳定性,研究了纳米豆粕在小鼠消化道内的分布情况。本实验采用双抗夹心ELISA定量分析纳米豆粕在小鼠消化道的分布情况,并与普通豆粕进行对比。研究结果显示:纳米豆粕、普通豆粕在消化道中的分布无显著性差异,同时也表现出相同的分布规律:灌胃0.5h之后两种豆粕主要分布在胃部,1h之后主要分布在小肠。此外纳米豆粕、普通豆粕、荧光微球在小鼠消化道各部位的分布比例也呈现出类似的规律:0.5h大部分分布在胃和小肠,在大肠中分布很少。在1-3.5h后分布最多的部位在小肠的空肠、回肠部位,大肠中的分布很少。通过对普通豆粕组和纳米豆粕组分布的显著性分析发现:在胃、十二指肠、空肠、回肠、直肠这些部位纳米豆粕的分布比例和普通豆粕无显著性差异(P0.05),而在结肠部位纳米豆粕组高于普通豆粕组(P0.05)。灌胃后在0.5h、1h、2h、3.5h,检测到消化道内的纳米豆粕占到灌胃总量的百分比分别为16.8%、23.4%、24.9%、24.6%,而普通豆粕占到灌胃总量的百分比分别为17.1%、23.6%、24.8%、24.9%。同样的时间段,消化道内的荧光占到灌胃总量的百分比分别为19.3%、29.7%、32.5%、33.3%。3.普通豆粕组和纳米豆粕组小鼠的消化道制作石蜡切片HE染色,结果表明给小鼠灌胃纳米豆粕3.5h后纳米豆粕组和普通豆粕组小鼠消化道形态结构相似。之后又做了免疫组化胶体金染色,结果发现:纳米豆粕在消化道中主要存在于小肠皱襞柱状细胞和大肠的细胞质中,个别出现在胃的黏膜层,局部的胶体金出现了聚集现象。
[Abstract]:In this paper, nano-soybean meal was taken as the research object, including the preparation and particle size characterization of nano-soybean meal, the distribution of nano-soybean meal in various parts of mouse digestive tract, and the effect of nano-soybean meal on the morphology and structure of mouse digestive tract. The main research methods, results and conclusions are as follows: 1. Ordinary soybean meal was prepared by ball milling. The morphology and particle size of nano-soybean meal were characterized by scanning electron microscope (SEM) and laser particle size analyzer. The results showed that the particle size distribution of soybean meal was 200 ~ 300 nm and its monodispersity was good. SEM observation showed that some of the nano-soybean meal particles had spherical morphology, some were oval, others were irregular shape, in which the proportion of sphericity and ellipse was larger, and the size distribution of nano-soybean meal particles was more uniform. The results of laser particle size analyzer show that the minimum particle size of soybean meal obtained by dry grinding is near 200nm, the particle size distribution is uniform, and the peak value of particle size distribution is at 219.8nm. In this paper, the distribution of nano-soybean meal in the digestive tract of mice was studied by using the antigenicity and relative stability of 尾-concomitant soybean globulin. The distribution of nano-soybean meal in the digestive tract of mice was quantitatively analyzed by double-antibody sandwich ELISA and compared with that of common soybean meal. The results showed that there was no significant difference in the distribution of nano-soybean meal and common soybean meal in the digestive tract. At the same time, the two kinds of soybean meal were mainly distributed in the stomach after 0.5 h and in the small intestine 1 hour later. In addition, the distribution of nano-soybean meal, common soybean meal and fluorescent microspheres in the digestive tract of mice showed a similar pattern: most of them were distributed in the stomach and small intestine at 0.5h, but little in the large intestine. The largest distribution was found in jejunum, ileum and large intestine after 1 ~ 3.5 h. The distribution of nano-soybean meal in stomach, duodenum, jejunum, ileum and rectum was not significantly different between ordinary soybean meal group and normal soybean meal group (P0.05), by analyzing the distribution of nano-soybean meal in common soybean meal group and nano-soybean meal group (P0.05). But in colon, nano-soybean meal group was higher than ordinary soybean meal group (P0.05). At 0.5 h, 1 h, 2 h, 3.5 h after gavage, the percentage of nano-soybean meal detected in the digestive tract was 16.8%, 23.4%, 24.9%, 24.6%, respectively. The percentage of common soybean meal was 17.1%, 23.6%, 24.8% and 24.9% respectively. Over the same period of time, the percentage of fluorescence in the digestive tract to the total gavage was 19.3%, 29.7%, 32.5%, 33.3% respectively. The paraffin-embedded sections of the digestive tract were stained with HE from the normal soybean meal group and the nano-soybean meal group. The results showed that the digestive tract morphology and structure of the nano-soybean meal group and the ordinary soybean meal group were similar to those of the ordinary soybean meal group after 3.5 h of feeding the mice with nano-soybean meal. The results showed that nano-soybean meal mainly existed in the columnar cells of the small intestine fold and the cytoplasm of the large intestine, some of them in the mucous layer of the stomach, and the colloidal gold in the local part of the stomach was aggregated. The results showed that nano-soybean meal mainly existed in the columnar cells of the small intestine and the cytoplasm of the large intestine.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S816;TS209

【参考文献】