不同环境温度下BlE对小鼠肠道黏膜保护和消化吸收的研究.docx
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1、分 类 号: 密 级: 学校代码: 学 号: 硕 士 学 位 论 文不同环境温度下BLE对小鼠肠道黏膜保护和消化吸收的研究论 文 作 者 XXX 指 导 教 师 XXX 教授 学 科 专 业 野生动植物保护与利用 申请学科门类 农学硕士 所 在 学 院 生命科学学院 论文提交日期 2020年4月 论文答辩日期 2020年5月 XXXX学位评定委员会XXXX2020年5月Study of BLE on intestinal mucosa protection and digestion and absorption of mice under different environment temp
2、eraturesA DissertationSubmitted to the Graduate FacultyIn Partial Fulfillment of the RequirementFor the Degree of Master of AgricultureByXXXSupervised byProfessor XXXMajor inWildlife Conservation and UtilizationInCollege of Life ScienceXXX UniversityXX, XX Province, ChinaMay, 2020本研究得到国家自然科学基金面上项目“竹
3、中植物次生代谢产物对大熊猫觅食策略和繁殖活动的影响”(31470462),国家林业局大熊猫保护国际合作项目“基于主食竹成分分析的大熊猫栖息地质量评价体系的建设”,XXX教育厅科研创新团队(自然科学)“大熊猫营养生态的研究”(17TD0037)项目资助!特此感谢!目 录目 录中文摘要VAbstractVII主要符号对照表X第1章 前言11.1 低温环境与恒温动物11.2 低温环境对动物生长发育的影响11.3 低温环境对动物肠道结构和功能的影响11.3.1 动物肠黏膜结构和功能21.3.2 低温环境对肠黏膜结构的影响21.3.3 低温环境对肠道消化吸收功能的影响31.4 外源添加物在低温环境下对肠
4、道的保护作用31.5 植物提取物研究41.5.1 植物提取物概述41.5.2 竹叶提取物概述51.5.3 巴山木竹叶概述51.6 研究目的及意义6第2章 不同环境温度下灌注BLE对小鼠生长发育的影响72.1 巴山木竹叶提取物的制备72.1.1 研究地概况72.1.2 样品采集与预处理72.1.3 仪器与试剂72.1.4 巴山木竹叶的提取72.2 试验动物的饲养82.2.1 试验材料82.2.2 试验动物82.2.3 试验设计82.2.3 检测指标与方法92.2.4 数据整理与分析92.3 结果与分析92.3.1 一般观察92.3.2 不同环境温度下灌注BLE对小鼠体增重的影响92.3.3 不同
5、环境温度下灌注BLE对小鼠摄食量的影响112.3.4 不同环境温度下灌注BLE对小鼠食物利用率的影响122.4 讨论13第3章 不同环境温度下灌注BLE对小鼠小肠形态结构的研究143.1 材料与方法143.1.1 试验材料143.1.2 试验动物与设计143.1.3 样品采集与保存143.1.4 检测指标与方法143.1.5 数据整理与分析153.2 结果与分析153.2.1 不同环境温度下灌注BLE对小鼠十二指肠形态结构的影响153.2.2 不同环境温度下灌注BLE对小鼠空肠形态结构的影响173.2.3 不同环境温度下灌注BLE对小鼠回肠形态结构的影响193.3 讨论21第4章 不同环境温度
6、下灌注BLE对小鼠回肠消化酶活性的研究234.1 材料与方法234.1.1 试验材料234.1.2 试验动物与设计234.1.3 样品采集与保存234.1.4 检测指标与方法234.1.5 数据整理与分析244.2 结果与分析244.2.1 不同环境温度下灌注BLE对小鼠回肠胰蛋白酶活性的影响244.2.2 不同环境温度下灌注BLE对小鼠回肠淀粉酶活性的影响254.2.3 不同环境温度下灌注BLE对小鼠回肠脂肪酶活性的影响254.3 讨论26第5章 不同环境温度下灌注BLE对小鼠结肠黏蛋白面积的研究285.1 材料与方法285.1.1 试验材料285.1.2 试验动物与设计285.1.3 样品
7、采集与保存285.1.4 检测指标与方法285.1.5 数据整理与分析295.2 结果与分析295.2.1 不同环境温度和灌注BLE下小鼠结肠AB-PAS染色显微观察295.2.2 不同环境温度下灌注BLE对小鼠结肠黏蛋白面积及比值的影响305.3 讨论32第6章 不同环境温度下灌注BLE对小鼠结肠短链脂肪酸含量的研究346.1 材料与方法346.1.1 试验材料346.1.2 试验动物与设计346.1.3 样品采集与保存346.1.4 检测指标与方法346.1.5 数据整理与分析356.2 结果与分析356.3 讨论37结 论39参考文献40致 谢i关于学位论文原创性的声明ii关于学位论文使
8、用授权的声明ii在学期间的科研情况iiiX摘 要摘 要本试验旨在研究秦岭佛坪巴山木竹叶提取物(Bamboo Leaves Extract,BLE)在不同环境温度下对小鼠生长发育、肠道黏膜保护和消化吸收功能的影响。试验采用22双因子重复试验设计(饲养温度:常温25和低温4;BLE灌注剂量:0 mg/kg BW和340 mg/kg BW),选取7周龄雌性昆明小鼠共240只,随机分为4个处理(常温对照组,NT+CON;常温BLE灌注组,NT+BLE;低温对照组,LT+CON;低温BLE灌注组,LT+BLE),每个处理6个重复,每个重复10只小鼠,试验周期为28天。试验分两个阶段进行,分别于14天和2
9、8天时就小鼠食物利用率、肠道组织形态学、肠道消化酶活性、肠道黏蛋白面积和短链脂肪酸含量进行相关测定,试验结果表明:1、与常温组相比,低温组小鼠体增重显著降低(P0.05),摄食量极显著增加(P0.01),食物利用率极显著降低(P0.01);与对照组相比,BLE组小鼠体增重显著增加(P0.05),摄食量显著降低(P0.05),食物利用率极显著提高(P0.01);低温下灌注BLE组小鼠摄食量显著降低(P0.05),食物利用率显著提高(P0.05)。结果表明灌注BLE在一定程度上可缓解因低温引起的小鼠生长发育减缓现象。2、与常温组相比,低温组小鼠十二指肠、空肠和回肠绒毛高度及十二指肠绒隐比显著降低(
10、P0.05),十二指肠隐窝深度显著增加(P0.05);与对照组相比,BLE组小鼠十二指肠和回肠绒毛高度显著提高(P0.05),空肠绒毛高度和绒隐比有提高的趋势,提高幅度最高分别达10.02%和3.29%;低温下灌注BLE组十二指肠绒毛高度显著提高(P0.05)。结果表明灌注BLE在一定程度上可恢复因低温导致的小鼠肠道黏膜结构损伤,增强肠道对营养物质的消化吸收。3、与常温组相比,低温组小鼠回肠胰蛋白酶和脂肪酶活性显著降低(P0.05),淀粉酶活性有降低趋势,降低幅度最高达9.54%;与对照组相比,BLE组胰蛋白酶和脂肪酶活性显著提高(P0.05),淀粉酶活性有提高的趋势,提高幅度最高达4.87%
11、;低温下灌注BLE组胰蛋白酶和脂肪酶活性显著提高(P0.05)。结果表明灌注BLE可缓解因低温引起的小鼠肠道消化酶活性抑制现象。4、与常温组相比,低温组小鼠结肠中性及混合性黏蛋白、酸性黏蛋白和黏膜层的面积极显著降低(P0.01);与对照组相比,BLE组黏蛋白和黏膜层面积均有增加的趋势,增高幅度最高分别达11.79%和14.55%;低温下灌注BLE组黏膜层面积极显著增加(P0.01),中性及混合性黏蛋白面积与黏膜层面积比值显著降低(P0.05)。结果表明灌注BLE在一定程度上可改善因低温导致的肠道肠壁变薄及黏蛋白分泌减少的现象。5、与常温组相比,低温组小鼠结肠乙酸和总酸含量极显著降低(P0.01
12、),丁酸的含量显著降低(P0.05);与对照组相比,BLE组丁酸和异丁酸的含量显著增加(P0.05),其余酸有增加的趋势,增加幅度最高可达43.83%。结果表明灌注BLE在一定程度上具有改善和增强肠道短链脂肪酸水平的作用。综上所述,低温可破坏小鼠肠道黏膜结构、抑制肠道消化酶活性、减少黏蛋白分泌和降低短链脂肪酸水平,影响肠黏膜结构的完整性和正常的消化吸收功能,从而减缓小鼠生长发育。灌注一定剂量的巴山木竹叶提取物可改善小鼠肠黏膜结构和功能,即保护肠道黏膜,增强肠道对营养物质的消化吸收,从而促进小鼠的生长发育。关键词:巴山木竹叶;竹叶提取物;生长发育;肠黏膜保护;消化吸收AbstractAbstra
13、ctThe purpose of this experiment was to study the effects of bamboo leaves extract of Bashania fargesii (Bamboo Leaves Extract, BLE) from Qinling Foping on the growth and development, intestinal mucosa protection, and digestion and absorption of mice at different environment temperatures. 22 double
14、factor repeated test design was adopted (feeding temperature: 25C at normal temperature and 4C at low temperature; BLE infusion dose: 0 mg/kg BW and 340 mg/kg BW). 240 7-week-old female Kunming mice were randomly divided into 4 treatments (normal temperature control group, NT + CON; normal temperatu
15、re bamboo leaves extract group, NT+BLE; low temperature control group, LT+CON; low temperature bamboo leaves extract group, LT+BLE). There were 6 replicates for each treatment, 10 mice for each replicate, and the experimental period was 28 days. The experiment was carried out in two stages, and food
16、 utilization, intestinal histomorphology, intestinal digestive enzyme activity, intestinal mucin area and short-chain fatty acid content of mice were measured at 14 and 28 days respectively. The test results showed that:1. Compared with the normal temperature group, the body weight gain of mice in t
17、he low temperature group was significantly reduced (P0.05), the food intake was significantly increased (P0.01), and the food utilization was significantly reduced (P0.01); compared with the control group, the body weight gain of the BLE group was significantly increased (P0.05), the food intake was
18、 significantly reduced (P0.05), and the food utilization was significantly increased (P0.01); at low temperature, the food intake of the BLE group was significantly reduced (P0.05), and the food utilization was significantly increased (P0.05). The results showed that the infusion of BLE can alleviat
19、e the growth and development slowdown of mice caused by low temperature to some extent.2. Compared with the normal temperature group, the villus height of duodenum, jejunum and ileum and VH/CD of duodenum in the low temperature group were significantly reduced, and the crypt depth of duodenum was in
20、creased (P0.05); compared with the control group, the villus height of duodenum and ileum of mice of the BLE group were significantly increased (P0.05), and it had a tendency to increase the villus height and VH/CD of jejunum, with the maximum increase of 10.02% and 3.29%, respectively; at low tempe
21、rature, the villus height of duodenum in the BLE group was significantly increased (P0.05). The results showed that the infusion of BLE can restore the damage of intestinal mucosal structure caused by low temperature in mice to a certain extent, and enhance the digestion and absorption of nutrients
22、in the intestinal tract.3. Compared with the normal temperature group, the ileum trypsin and lipase activity of mice in the low temperature group were significantly reduced (P0.05), and the amylase activity showed a decreasing trend, with the highest decreasing range of 9.54%; compared with the cont
23、rol group, the activity of trypsin and lipase in the BLE group was significantly increased (P0.05), and the amylase activity had a tendency to increase, with a maximum increase of 4.87%; the activity of trypsin and lipase in the BLE group was increased significantly at low temperature (P0.05). The r
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