沙棘蛋白对糖尿病小鼠肠道微生物及脂类代谢的影响

发布时间：2018-03-17 10:51

本文选题：沙棘蛋白　切入点：2型糖尿病　出处：《合肥工业大学》2017年硕士论文　论文类型：学位论文

【摘要】：沙棘(Hippophae rhamnoides Linn.)植物沙棘为胡颓子科(Elaeagnaceae)沙棘属(Hippophae),是一种落叶性灌木,为我国西部地区最具代表性的经济作物之一。其生物资源蕴藏量丰富,多样性广泛。沙棘为药食同源植物,它的叶、茎、根和果,特别是沙棘的果实含有丰富的生物活性物质和营养物质。其中,从沙棘的籽中提取所得的蛋白,含量丰富,氨基酸种类齐全,含有人体所需的全部必需氨基酸,是一种优良的植物蛋白资源。本文以沙棘籽为原料,采用酸沉碱提的方法来制备沙棘籽蛋白。利用高脂饮食结合链脲佐菌素(STZ)来诱导,获得2型糖尿病动物实验模型。使用沙棘蛋白进行灌胃处理小鼠4周,探究其对小鼠体重(BW)、空腹血糖水平(FBG)、血脂水平和肝脏损伤的影响,用于研究沙棘蛋白对糖尿病小鼠的降血脂作用;检测各个时期小鼠粪便的pH值和肠道优势菌群的数量变化,考察沙棘籽蛋白对糖尿病小鼠肠道环境和肠道菌群的影响;同时用半定量PCR方法(RT-PCR)研究对脂肪酸代谢mRNA表达水平的影响,初步探究沙棘籽蛋白改善糖尿病脂质转运机理。利用16srDNA测序技术对小鼠粪便全基因组进行分离分类,考察沙棘蛋白对糖尿病小鼠肠道微生物菌群的动态调节。实验结果显示,采用碱提酸沉的工艺制备沙棘籽蛋白,其含量为78.6%,总提取率为37.4%。动物实验的结果显示:沙棘蛋白处理组较模型组小鼠的体重、空腹血糖、总甘油三酯(TG)、总胆固醇(TC)和低密度脂蛋白胆固醇(LDL-C)与糖尿病组相比有显著降低(p0.05);高密度脂蛋白胆固醇(HDL-C)较糖尿病组则明显升高。此外,RT-PCR结果显示,沙棘籽蛋白使小鼠肝脏组织基因CCATT增强子结合蛋白(CEBPα)、脂肪酶合酶(FAS)、乙酰辅酶A羧化酶(ACC1),肉碱酰基转移酶(CPT1)的表达量增加,降低胰岛素抵抗,改善糖尿病小鼠症状。培养得到的肠道微生物中,沙棘蛋白处理组的双歧杆菌(Bifidobacterium)、乳酸杆菌(Lactobacillus)、拟杆菌(Bacteroides)较模型组均有显著的增加;球形梭菌(Clostridium coccoides)则明显减少;聚类分析结果提示沙棘蛋白处理组肠道菌群与正常组之间的相似性较高,达77%左右。本研究表明:沙棘籽蛋白能改善2型糖尿病高血糖、高血脂;对肠道微生物同样有显著的调节作用。实验证明沙棘籽蛋白能够提高脂肪酸的代谢。PCRDGGE的分析结果表明沙棘籽蛋白能够显著改善糖尿病小鼠肠道菌群环境,调节动态平衡,进而改善2型糖尿病症状。
[Abstract]:Hippophae rhamnoides Linn.) Hippophae is a deciduous shrub of the genus Elaeagnaceae.Hippophae is one of the most representative cash crops in western China. Its leaves, stems, roots and fruits, especially the fruits of Hippophae rhamnoides, are rich in bioactive substances and nutrients. Hippophae rhamnoides seed was prepared by acid precipitation and alkali extraction, and was induced by high fat diet combined with streptozotocin (STZ). The experimental animal model of type 2 diabetes mellitus was established. The effects of sea buckthorn protein on body weight, fasting blood glucose level, lipid level and liver injury were investigated in mice treated with sea buckthorn protein for 4 weeks. To study the hypolipidemic effect of seabuckthorn protein on diabetic mice, to detect the pH value of feces and the number of dominant microflora of intestinal tract in each period of mice, and to investigate the effect of seabuckthorn seed protein on the intestinal environment and intestinal flora of diabetic mice. At the same time, the effect of RT-PCR on fatty acid metabolism mRNA expression was studied by semi-quantitative PCR method, and the mechanism of seabuckthorn seed protein (Hippophae rhamnoides) improving lipid transport in diabetes mellitus was preliminarily explored. The whole genome of mouse feces was separated and classified by 16s rDNA sequencing technique. The dynamic regulation of seabuckthorn protein on intestinal microflora of diabetic mice was investigated. The results showed that the process of alkali extraction and acid precipitation was used to prepare seabuckthorn seed protein (Hippophae rhamnoides L.). Its content was 78.6 and the total extraction rate was 37.4. The results of animal experiments showed that the body weight and fasting blood glucose of mice treated with Hippophae rhamnoides protein were higher than those of model group. Total triglyceride (TGG), total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) were significantly lower than those in diabetic group (P 0.05), and HDL-C (high density lipoprotein cholesterol) was significantly higher than that in diabetes mellitus (DM). Hippophae rhamnoides seed protein (Hippophae rhamnoides) increased the expression of CCATT enhancer binding protein (CEBP 伪), lipase synthase (FASA), acetyl coA carboxylase (ACC1), carnitine transferase (carnitine transferase), and decreased insulin resistance. To improve the symptoms of diabetic mice. In the intestinal microbes cultured with Hippophae rhamnoides, Bifidobacterium teriumus, Lactobacillus lactis, Bacteroidesin treated with seabuckthorn protein were significantly increased compared with the model group, while Clostridium coccoidesin was significantly decreased. Cluster analysis showed that the similarity between the intestinal flora of seabuckthorn protein treated group and normal group was about 77%. The results showed that Hippophae rhamnoides seed protein could improve hyperglycemia and hyperlipidemia in type 2 diabetes mellitus. The results of PCRDGGE analysis showed that seabuckthorn seed protein could significantly improve the intestinal microflora environment and regulate the dynamic balance of diabetic mice. And then improve the symptoms of type 2 diabetes.
【学位授予单位】：合肥工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.1

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