【作者】 李蕾；

【导师】 张晶； 徐暾海；

【作者基本信息】 吉林农业大学 ， 中药学， 2015， 硕士

【摘要】 刺萼悬钩子(Rubus alexeterium Focke)为蔷薇科悬钩子属植物,国内外未见有刺萼悬钩子的相关报道,本论文首次对刺萼悬钩子枝化学成分、不同提取部分降糖活性筛选以及总黄酮含量进行了研究,并对有效部位作用机制进行初步研究,为新型糖尿病药物的研究提供参考。通过检索、归纳相关文献,本论文对悬钩子属植物及刺萼悬钩子的主要的化学成分和药理活性进行了综述,为刺萼悬钩子的进一步研究提供参考。本论文采用不同极性的溶剂对刺萼悬钩子枝进行渗漉提取,得到石油醚提取部分、乙酸乙酯提取部分、95%乙醇提取部分、70%乙醇提取部分和30%乙醇提取部分。采用超声提取法对刺萼悬钩子枝残渣进行超声提取,得到水提取部分,分别用大孔吸附树脂和聚酰胺对水提液进行吸附,用不同浓度的醇进行洗脱,共得到6个提取部分。采用高速逆流色谱法对乙酸乙酯提取部分和95%乙醇提取部分进行最佳分离条件摸索,最终得到高速逆流色谱法分离乙酸乙酯提取部分的最佳条件为正己烷:乙酸乙酯:甲醇:水=5:4:3.5:1,高速逆流色谱法分离95%乙醇提取部分的最佳条件为氯仿:正丁醇:甲醇:水=4:0.5:3:2。本论文研究了刺萼悬钩子枝石油醚提取部分、乙酸乙酯提取部分、95%乙醇提取部分、70%乙醇提取部分和30%乙醇提取部分对KKAy糖尿病小鼠体重、空腹血糖、饮食量、饮水量、胰岛素耐量以及生化指标等的干预作用,通过综合统计分析可知,乙酸乙酯提取部分和30%乙醇提取部分具有较好的作用。乙酸乙酯提取部分可使小鼠体重显著下降;与模型组比较,乙酸乙酯提取部分对小鼠体重、肾体比的影响具有显著性差异。乙酸乙酯提取部分和30%乙醇提取部分可使小鼠饮水量、饮食量以及空腹血糖下降,同时乙酸乙酯提取部分和30%乙醇提取部分可使血清生化指标TG、TC和LDL含量下降,使HDL的含量升高。本论文采用硅胶色谱、葡聚糖凝胶和半制备型高效液相色谱相结合的方法对有效部位乙酸乙酯提取部分进行分离纯化,得到7个化合物,最终鉴定出5个化合物,分别为邻苯二甲酸二异丁酯、岩白菜素、β-谷甾醇、3,4-二羟基苯甲酸(又名原儿茶酸)、熊果酸,未鉴定出的两种化合物为三萜类化合物,可以对鉴定出的化合物进行进一步的药理活性及作用机制研究。本论文采用Realtime PCR法研究了乙酸乙酯部位对KKAy小鼠肌肉组织中五种基因表达量的影响,可知乙酸乙酯部位可使小鼠肌肉组织中pparαmRAN和pparγmRAN两种基因表达量升高,使小鼠肌肉组织glut4 mRAN、mnsod mRAN和ampk1 mRAN三种基因表达量降低。本论文采用紫外分光光度法对刺萼悬钩子枝中总黄酮含量进行测定,考察了乙醇体积分数、料液比、超声时间和超声次数对植物中总黄酮提取率的影响,通过正交试验和节能观念综合考虑,确定刺萼悬钩子枝总黄酮提取的最优工艺条件为乙醇体积分数50%,料液比1:40,超声功率100W,超声时间60min,超声次数2次。本文创新点:本文首次对刺萼悬钩子枝进行研究。更多还原

【Abstract】 Rubus alexeterium Focke belongs to the Rosaceae rubus, and there was no relevant reports at home and abroad about Rubus alexeterium Focke. This paper focuses on the research of chemical composition 、 the screening of the different extraction parts of hypoglycemic activity and total flavonoids content study of the branch from Rubus alexeterium Focke for the first time, and preliminaried studied of its mechanism, which provided the reference basis for the research of new diabetes drugs.By retrieving and summarizing lots of literatures, this paper reviewed the Rubus and Rubus alexeterium Focke main chemical components and pharmacological activity, providing a reference basis for further study on Rubus alexeterium Focke.This thesis adopted different polar solvent to percolate extraction Rubus alexeterium Focke branch, and had been extracted with petroleum ether、ethyl acetate extracts、95% ethanol extract、70% ethanol extract and 30% ethanol extracts. Adopting ultrasonic extraction to extract the Rubus alexeterium Focke branch residue, got water extracted part. Respectively using macroporous adsorption resin and polyamide to adsorb water extraction, got 6 extraction sites. The paper groped for the best separation conditions by high speed counter current chromatography in ethyl acetate extract and 95% ethanol extract. The optimum conditions for separation of ethyl acetate part of high speed countercurrent chromatography was n-hexane: ethyl acetate: methanol: water =5:4:3.5:1,and the optimum conditions for separation of 95% ethanol extract of high speed countercurrent chromatography was chloroform: n-butanol: methanol: water =4:0.5:3:2.This paper studyed the intervention effect of Rubus alexeterium Focke branch with petroleum ether、ethyl acetate extract、95% ethanol extract、70% ethanol extract and 30% ethanol, on the part of KKAy diabetic mice body weight、fasting blood sugar、diet and water intake、insulin tolerance and the biochemical indicators. Through the comprehensive statistical we knowed that the ethyl acetate extract and 30% ethanol extract had a good effect. Ethyl acetate extracts could cause the decrease of body weight of mice. Compared with model group, the ethyl acetate extract had significant difference on mice body weight and kidney ratio. The ethyl acetate extract and 30% ethanol extract could make mice diet and water intake and the amount of the fasting blood glucose decreased. At the same time, ethyl acetate extract and 30% ethanol extract could make the biochemical indicators of TG, TC and LDL content decrease, and HDL content increase.Using the method of silica gel column chromatography 、 Sephadex and semi preparative HPLC coupled to separate and purify the ethyl acetate extract, got seven compounds. Finally five compounds had been identified, they respectively were dibutyl phthalate 、bergeninum、β-sitosterol、3,4-dihydroxy benzoic acid(Protocatechuic acid)、ursolic acid. There was two kinds of three terpenes which was not identified was three terpenoids, which can be studied pharmacological activities and mechanism for the further.Using Realtime PCR method to study the influence of the ethyl acetate extract on the amount of five gene of muscle tissue in mice with KKAy PPAR alpha. The ethyl acetate extract could make the muscle tissue of mice pparα mRAN and PPARγ mRAN two gene expression increase, and make the muscle tissue of mouse glut4 mRAN、mnsod mRAN and ampk1 mRAN three gene expression decrease.This paper usd UV spectrophotometry to study the total flavonoids content of Rubus alexeterium Focke branch, and investigated the effects of ethanol concentration、 ratio of material to liquid ratio、 ultrasonic time and ultrasonic frequency on total flavonoids in plant extract. By orthogonal test and the idea of saving energy comprehensive consideration, determine the optimal process conditions of Rubus alexeterium Focke branch total flavone extraction was ethanol concentration 50%, ratio of solid to liquid 1:40, ultrasonic power 100 W, ultrasonic time 60 min, ultrasonic frequency 2 times. The innovation of this paper:The study of Rubus alexeterium Focke branch was the first time.更多还原