天然产物虾青素抗氧化与降糖作用及其部分机制研究

发布时间：2018-07-27 11:36

【摘要】：[目的]优化雨生红球藻有机溶剂提取工艺,寻找最佳的萃取条件,精确检测雨生红球藻萃取物中的虾青素酯含量,探索适用于大规模工业生的高速、高效、低成本提取方法;建立小鼠糖尿病模型,并给予不同浓度的雨生红球藻源虾青素酯,研究其对糖尿病小鼠糖耐量的影响、抗氧化指标的影响、血脂的调节情况及胰腺组织的损伤修复情况。探讨虾青素酯改善糖尿病及其并发症的作用机制,并验证高剂量虾青素酯对正常小鼠的影响;结合市场需求及原料特性,制备以雨生红球藻萃取物为原料的功能食品。[方法]1.从原料的预处理入手,采用物理、化学处理相结合的手段,首先将雨生红球藻微藻进行破壁粉碎,以食品级有机溶剂乙酸乙酯作为萃取剂,应用SPSS 22.0统计学软件,对萃取温度(40℃、50℃、60℃)、原料颗粒度(40目、80 目、160 目)、料液比(1:4、1:8、1:16)、萃取时间(30min、60min、90min)设计正交实验,并对单因素水平进行验证,通过与仲裁法丙酮萃取得率作对比,结合实际操作,确定雨生红球藻最佳萃取因素水平。2.运用高效液相色谱法(High Performance Liquid Chromatography HPLC),精确检测雨生红球藻萃取物中虾青素酯的含量。3.将喂食30 d高剂量虾青素酯正常小鼠,与喂食30 d溶剂的正常小鼠血糖值做对比,用SPSS 22.0统计学软件进行分析,比较两组小鼠的血糖值,分析虾青素酯对正常小鼠血糖值的影响。4.采取四氧嘧啶腹腔注射的方式建立糖尿病小鼠模型,设立高中低三个剂量虾青素酯组和二甲双胍、维生素C两个阳性药对照组,给药期周期为30 d,每7 d记录1次小鼠体重;给药期末进行耐糖量实验,利用血糖仪测定血糖值,运用全自动生化分析仪和相关试剂盒检测丙二醛(MDA)、谷胱甘肽(GSH-Px)、超氧化物歧化酶(T-SOD)等抗氧化指标,总胆固醇(CHO)、甘油三酯(TG)等血脂指标,并制作胰腺病理切片,观察胰岛组织形态变化。5.根据原料理化性质,选用软胶囊作为虾青素酯功能食品最终产品剂型,并进行小试试验研究生产。[结果]1.萃取温度40℃,原料颗粒度160目,料液比1:16,萃取时间90min,此条件下雨生红球藻萃取效果最好,萃取率高达90.8%(W/W)。2.通过HPLC分析,确定雨生红球藻萃取物中虾青素酯含量为5.46%。3.高剂量虾青素酯喂养30 d,对正常小鼠的血糖无影响。4.四氧嘧啶造模后小鼠毛色杂乱暗淡,出现明显多饮、多食、多尿的"三多"症状,与正常组小鼠相比,体重明显减轻(P0.05)。给予不同剂量虾青素酯喂养30 d后,各给药组体重平均值与模型对照组相比有所上升,低剂量虾青素酯组小鼠体重升高明显,存在统计学差异(P0.05)。高、中、低剂量虾青素酯组的空腹血糖值有不同程度下降,与模型对照组比较有统计学差异(P0.05,P0.05,P0.05),且有一定的量效关系,但降血糖效果不如阳性对照药二甲双胍。在口服葡萄糖耐量实验中,各组小鼠的糖负荷均在30 min时达到峰值,高剂量虾青素酯组和中剂量虾青素酯组血糖值在120 min时明显低于对照组(P0.05,p0.05),高、中剂量虾青素酯可以提高糖尿病小鼠对外源性葡萄糖的耐受力,与对照组相比有统计学差异。计算血糖线下面积,高、中、低剂量虾青素酯组均比模型对照组低,且有统计学意义(P0.05,P0.05,P0.05),表明虾青素酯可降低各时间点血糖,但效果不如阳性对照药二甲双胍。抗氧化指标,与模型对照组相比,高、中、低剂量虾青素酯组血清MDA含量显著下降(P0.01,P0.01,P0.05),血清GSH-Px活性单位含量、血清T-SOD活性单位含量显著上升(P0.01,P0.01,P0.01),并存在着一定的量效关系,抗氧化指标结果表明:虾青素酯具有显著的抗氧化作用,且明显高于阳性对照药维生素C。虾青素各剂量组血清CHO、TG含量明显低于对照组,差异具有统计学意义(P0.01,P0.01,P0.01),提示虾青素酯具有调节血脂的作用。5.以雨生红球藻萃取物为主要原料,葵花籽油为辅料,制成内容物0.5 g的虾青素软胶囊。
[Abstract]:[Objective] to optimize the extraction process of the organic solvent of rhodotrazina, find the best extraction conditions, accurately detect the content of astaxanthin ester in the extract of rhodotrae alga, explore the high speed, high efficiency and low cost extraction method suitable for large-scale industrial students, establish a diabetic model of mice, and give astaxanthin esters of different concentrations of the raining red ball algae. The influence of the glucose tolerance, the influence of antioxidant index, the regulation of blood lipid and the injury and repair of pancreatic tissue were studied. The mechanism of astaxanthin ester to improve diabetes and its complications was investigated, and the effect of high dose astaxanthin ester on normal mice was tested and the effect of astaxanthin ester on normal mice was verified. [method]1., starting with the pretreatment of the raw material, starting with the pretreatment of raw materials, using the means of physical and chemical treatment, the rain raw red alga microalgae were broken, and the organic solvent ethyl acetate was used as the extractant and the SPSS 22 system software was applied to the extraction temperature (40, 50, 60), and the raw materials were used. Granularity (40 orders, 80 orders, 160 orders), material liquid ratio (1:4,1:8,1:16), extraction time (30min, 60min, 90min) design orthogonal experiment, and verify the single factor level, by comparing with the arbitration method of acetone extraction, combining actual operation, to determine the optimum extraction factor level of raococcus red pellet using high performance liquid chromatography (High Performance L) method (High Performance L). Iquid Chromatography HPLC), the content of astaxanthin ester in the extract of rhodotrae alga was accurately measured by.3., 30 d high dose astaxanthin ester was fed to normal mice, and the normal mice fed with 30 d solvent were compared. The blood glucose of two groups of mice was compared with the SPSS 22 statistical software, and the blood of astaxanthin was compared to the normal mice blood. The effect of sugar value on.4. was established by intraperitoneal injection of four pyrimidine to establish a diabetic mouse model. Three doses of astaxanthin ester group and metformin and two vitamin C positive drug control groups were set up in high school. The period of the administration period was 30 d, and 1 mice were recorded every 7 d; the glucose tolerance test was carried out at the end of the period and the blood glucose was measured by blood glucose meter, The antioxidant indexes such as malondialdehyde (MDA), glutathione (GSH-Px), superoxide dismutase (T-SOD), total cholesterol (CHO), triglyceride (TG) and other lipid indexes were detected by the automatic biochemical analyzer and related kit. The pathological sections of pancreas were made and the morphological changes of pancreatic islets were observed and.5. was used as the shrimp green according to the properties of raw material. The final product form of vegetarian ester function food was produced in a small test. [results]1. extraction temperature 40 C, raw material granularity 160 mesh, material liquid ratio 1:16, extraction time 90min, the extraction efficiency is best, the extraction rate is up to 90.8% (W/W).2. through HPLC analysis, determine the astaxanthin ester content in the raining red pellet extract is 5 .46%.3. high dosage of astaxanthin ester was fed to 30 d, and the blood sugar of normal mice had no effect on the hair color and confusion of mice after the model of.4. four oxacil. The "more than 3" symptoms of polyuria, polyuria and polyuria were obviously reduced (P0.05). The average weight and model of body weight of each group were given after 30 d were fed with different doses of astaxanthin ester. Compared with the control group, the body weight of the low dose astaxanthin ester group was significantly higher than that of the low dose astaxanthin ester group (P0.05). High, middle and low dosage of astaxanthin ester group decreased the fasting blood glucose in different degrees, compared with the model control group, there was a statistical difference (P0.05, P0.05, P0.05), and there was a certain dose effect relationship, but the effect of hypoglycemic effect was not as good as that of Yang. In the oral glucose tolerance test, in the oral glucose tolerance test, the glucose load reached the peak at 30 min. The high dose of astaxanthin ester group and the middle dose astaxanthin ester group were significantly lower than the control group (P0.05, P0.05) at 120 min. High dosage of astaxanthin ester could improve the tolerance to exogenous glucose in diabetic mice. The area under the blood glucose line, high, middle and low dosage of astaxanthin ester group were lower than the model control group, and there were statistical significance (P0.05, P0.05, P0.05), indicating that astaxanthin ester could reduce the blood glucose at all time points, but the effect was not as effective as the positive metformin. The antioxidant index was compared with the model control group. The content of serum MDA in high, middle and low dose astaxanthin ester group decreased significantly (P0.01, P0.01, P0.05), the content of GSH-Px active unit in serum, the content of serum T-SOD active unit increased significantly (P0.01, P0.01, P0.01), and there was a certain quantitative effect relationship. The antioxidant index showed that astaxanthin ester had significant antioxidant effect and was obviously higher than positive. The content of serum CHO and TG in each dose group of vitamin C. and astaxanthin was significantly lower than that of the control group. The difference was statistically significant (P0.01, P0.01, P0.01), suggesting that astaxanthin ester had the effect of regulating blood lipids,.5. was the main raw material, and sunflower seed oil was used as the auxiliary material, and the astaxanthin soft capsule of 0.5 g was made.
【学位授予单位】：中央民族大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TS201.4

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