Anti-oxidation And Hypoglycemic Effects Of Natural Product Astaxanthin And Its Partial Mechanism

Objective:To optimize the extraction process of Haematoccus pluvialis,looking for the best extraction method by organic solvent.Accurate measurement of Haematococcus pluvialis astaxanthin esters in the organic solvent extract.explore the high speed,high efficiency and low cost industrial production application of Haematococcus pluvialis extraction method;Establishment of diabetic mice model,treated with different concentrations of Haematoccus pluvialis source of astaxanthin esters,To study the effect of it on Oral Glucose Tolerance Test,antioxidant indexes,blood lipid regulation and pancreatic tissue damage repair of diabetic mice.To investigate the mechanism of astaxanthin ester effect on diabetes and its complications,and to verify the effect of high dose astaxanthin esters of normal mice;Combined with the market demand and the characteristics of raw materials,functional food preparation in Haematococcus pluvialis extract as raw material.Method:1.From the pretreatment of raw material,wall-brokenad haematococcus pluvialis by the means of the combination of physical and chemical.Use food grade organic solvents of ethyl acetate as the extraction agent,using SPSS 22.0 statistical software,designthe orthogonal experimental with extraction temperature(40 ?,50?and 60 ?),particle size(40 mesh,80 and 160 mesh),solid-liquid ratio(1:4,1:8,1:16),extraction time(30 min,60 min,90 min),and verify the single factor,the yield compared with the arbitration law and the use of acetone extraction,accurate analysis by HPLC,to determine the optimum extraction parameters of Haematoccus pluvialis.2.Based on the analysis of the content of carotenoids in the Haematococcus pluvialis,the High Performance Liquid Chromatography(HPLC)method was used to determine the content of astaxanthin esters in the extract of Haematococcus pluvialis.3.Fed 30 d high dose astaxanthin ester in normal mice,compared with normal mice fed 30 d of solvent,were analyzed by SPSS 22.0 statistical software,compare two groups of mice blood glucose,verify the effect of astaxanthin ester on blood glucose of normal mice.4.Intraperitoneal injection of Alloxan to establish diabetic mice model,set up three intervendonal astaxanthin ester group and metformin,vitamin C two positive drug control group,administered during the period of 30 d,every 7 d to record the weight.Oral Glucose Tolerance Test was performed at the end of medication and blood glucose was measured,utilization blood glucose meter.automatic biochemical analyzer and related kit for detection of antioxidant index as MalonDialdehyde(MDA),Glutathione Peroxidase(GSH-Px),Total Superoxide Dismutase(T-SOD)and,lipids as total cholesterol(CHO),triglyceride(TG).and pancreatic pathology,observe the morphological changes of pancreatic tissue.5.According to the Physical and chemical properties of raw materials,soft capsule was selected as the final product of astaxanthin ester functional food.Small scale production.Results:1.When extraction temperature is 40?,the particle size of 160 mesh,solid-liquid ratio 1:16,extraction time 90 min,the Haematoccus pluvialis extraction effect was the best extraction rate as high as 90.8%(W/W).2.Through the HPLC analysis,the content of astaxanthin esters in the extract was determined to be 5.46%.3.High dose astaxanthin esters had no effect on blood glucose of normal mice.4.Model mice appear hair disorder and dim,appear classical symptoms,compared with normal mice,significant weight loss(P<0.05).Given different doses of astaxanthin esters after 30 d,the average weight in each group increased compared with the model control group,the weight of low dose astaxanthin esters group mice increased significantly,there is a statistically significant difference(P<0.05).High,low dose astaxanthin ester group fasting blood glucose value decreased in different degrees,compared with model control group have a statistically significant difference(P<0.05,P<0.05,P<0.05),and there is a dose-response relationship,but the hypoglycemic effect less than the positive control drug metformin.In Oral Glucose Tolerance Test,glucose load of mice reached the peak at 30min,high dose group and medium dose astaxanthin ester group astaxanthin ester glucose in 120min was significantly lower than the control group(P<0.05,P<0.05),high dose astaxanthin ester can improve the diabetic mice exogenous glucose tolerance.Calculate the area under the line,high blood glucose,medium and low dose group of astaxanthin esters higher than the model control group,and there was statistical significance(P<0.05,P<0.05,P<0.05),showed that astaxanthin esters can reduce the blood glucose of each time point,the effect is not as good as the positive control drug metformin.The antioxidant index,compared with the model group,high,middle and low dosage of astaxanthin ester group serum MDA content was significantly decreased(P<0.01,P<0.01,P<0.05),significantly increased the activity of serum GSH-Px content,serum T-SOD activity per unit content(P<0.01,P<0.01,P<0.01),both in a certain dose effect relationship,antioxidant index results show that astaxanthin ester has a significant antioxidant effect,and was significantly higher than the positive control drug vitamin C.The contents of CHO and TG in serum of astaxanthin group were significantly lower than those of control group,the difference was statistically significant(P<0.01,P<0.01,P<0.01).5.The soft capsules main raw material was extracted from the Haematococ-cus pluvialis,Sunflower seed oil as accessories,Content 0.5 g.