【作者】 杜道辉；

【导师】 刘亚明；

【作者基本信息】 湖北中医学院 ， 中西医结合基础， 2009， 硕士

【摘要】 目的:运用现代生物学技术,充分体现中医“治未病”的理论优势,利用奶牛的生物转化功能从富铬酵母中转化含高活性葡萄糖耐量因子(GTF)的功能牛奶,用中药活性成分葛根素组合制备而成GTF葛根功能牛奶,观察GTF葛根功能牛奶对葡萄糖耐量低减(IGT)小鼠的干预作用,通过对骨骼肌GLUT-4基因表达的影响来阐明其干预IGT的分子机制,为中西医结合方法干预治疗IGT提供新的思路。方法:(1)将奶牛随机分成四组,在奶牛饲料中分别添加不同剂量组的富铬酵母,通过检测牛奶中铬元素含量,观察牛奶铬含量与富铬酵母添加剂量的相关性。(2)采用D-半乳糖腹腔注射法诱导小鼠IGT模型,将GTF功能牛奶、普通牛奶、富铬酵母和二甲双胍分组灌胃,比较GTF功能牛奶与对照各组对IGT小鼠的干预作用。(3)从野葛中提取葛根素,与GTF功能牛奶按比例配制GTF葛根功能牛奶,初步完成GTF葛根功能牛奶的制备工艺。(4)采用D-半乳糖腹腔注射法诱导小鼠IGT模型,将GTF葛根功能牛奶按不同剂量分组灌胃,观察各组对IGT小鼠空腹血糖(FBG)、口服葡萄糖耐量试验(OGTT)、胰岛素耐量试验(ITT)、血清空腹胰岛素(FINS)和胰岛素抵抗指数(HOMA-IR)水平的影响。(5)采用逆转录-聚合酶链反应(RT-PCR)技术,通过检测经GTF葛根功能牛奶不同剂量组干预治疗后IGT小鼠骨骼肌组织葡萄糖转运蛋白4(GLUT-4)基因表达水平,来阐释GTF葛根功能牛奶对IGT小鼠干预作用的可能机制。结果:(1)牛奶铬含量与奶牛饲料中富铬酵母添加剂量两者高度相关。(2)GTF功能牛奶对IGT小鼠的干预作用与普通牛奶、富铬酵母组比较明显增强。(3)GTF葛根功能牛奶对IGT小鼠空腹血糖(FBG)无明显影响,降低口服葡萄糖耐量试验(OGTT)后2小时血糖(2hBG),增加胰岛素耐量试验(ITT)后胰岛素的敏感性,降低空腹胰岛素(FINS)和胰岛素抵抗指数(HOMA-IR)水平。(4)GTF葛根功能牛奶明显增加IGT小鼠骨骼肌组织葡萄糖转运蛋白4(GLUT-4)基因表达水平。结论:(1)饲料中添加的铬可经奶牛机体生物转化为高活性铬富集在牛奶中,牛奶中铬含量与饲料中铬含量呈正相关。(2)功能牛奶中的铬可能与牛奶中的多种氨基酸络合后形成葡萄糖耐量因子(GTF),调节葡萄糖代谢,对IGT小鼠起到干预作用。(3)葛根素与功能牛奶中的GTF组合后,药效上产生协同作用。(4)GTF葛根功能牛奶对IGT小鼠干预治疗的作用机制可能是通过增加外周靶细胞GLUT-4mRNA的表达,增强组织对葡萄糖的摄取,提高胰岛素敏感性从而干预治疗IGT的。更多还原

【Abstract】 Objective: To study theory advantages in the Chinese medicine "Preventive Treatment of Disease" , using modern biological technology, utilize the biotransformation function of cows transforming the function of milk with high-activity glucose tolerance factor (GTF) form chromium-enriched yeast. Preparating GTF Puerarin function milk form Chinese medicine active ingredients Puerarin, to observe the role of intervention to the function of GTF Puerarin milk on Grape impaired glucose tolerance (IGT) in mice, through skeletal muscle GLUT-4 gene expression to clarify its molecular mechanism of intervention on IGT, to provide new ideas for traditional Chinese and western medicine in the treatment of IGT intervention.Methods: (1) Dividing randomly the cows into four groups, to add the chromium-rich yeast to dairy cattle feed at different dose groups respectively, through the detection of chromium content in milk to conclude the relationship between milk chromium content and chromium-rich yeast additive dosage.(2)Choosing GTF functions milk, ordinary milk, chromium-rich yeast and metformin to lavage stomatch to impaired glucose tolerance (IGT) mice model induced by D-galactose peritoneum injection, to compare the intervention role on the IGT mice between the milk GTF functions and each control group.(3) Extracting Pueraria lobata Puerarin form wilding Puerarin with GTF function of milk in proportion to the preparation of GTF functions Puerarin milk, to achieve initially GTF Puerarin functional milk production process.(4)Adopting D-galactose in mice induced by intraperitoneal injection of IGT model, GTF Puerarin milk will function by gavage administration in different doses, to observe each group of mice IGT fasting blood glucose (FBG),oral glucose tolerance test (OGTT), insulin tolerance test (ITT),serum fasting insulin (FINS) and insulin resistance index (HOMA-IR) levels.(5)Applying Reverse transcriptase-polymerase chain reaction (RT-PCR) technology,though detecting Puerarin by GTF function of different doses of milk after treatment IGT intervention group of mice skeletal muscle glucose transporter 4 (GLUT-4) gene expression level, to explain the possible mechanism of GTF Puerarin functional milk interfere role in IGT mice.Results: (l)There were significant correlations between Chromium content of milk and volume of dairy feed additive chromium-rich yeast.(2) Compared with ordinary milk group, chromium-rich yeast group, GTF functions milk intervention on the role of IGT in mice increased.(3)GTF Puerarin functional milk of mice IGT fasting blood glucose (FBG) had no significant effect, reduced oral glucose tolerance test (OGTT) 2 hours after glucose (2hBG), increased insulin tolerance test (ITT) after the insulin sensitivity, lowered fasting insulin (FINS) and insulin resistance index (HOMA-IR) levels.(4)GTF Puerarin functional milk IGT mice significantly increased skeletal muscle glucose transporter 4 (GLUT-4) gene expression level.Conclusion: (1)Chromium diet dairy organism biology may be translated into highly active chromium enrichment in the milk, the milk in the chromium content and chromium content of feed was positively correlated.(2) Function of milk in the milk may be associated with chromium in a variety of amino acids formed after complexation glucose tolerance factor (GTF), regulation of glucose metabolism on the IGT intervention played a role in mice.(3) After Puerarin and function of milk composed with GTF assembling, the efficacy had a synergistic effect.(4)The role of mechanisms GTF Puerarin functional milk intervention on the treatment of IGT in mice was possible increasing the peripheral target cells the expression of GLUT-4mRNA to enhance the uptake of glucose Organize, improved insulin sensitivity.更多还原