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Research of Glycosaminoglycan from Perna Virids Effect on Blood Lipid

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Tutor: WuHongMian
School: Guangdong Ocean University
Course: Aquatic Products Processing and Storage Engineering
Keywords: Perna virids,Glycosaminoglycan,Purification,Hypolipidemic,Lipid metabolism,Antio
CLC: R285.5
Type: Master's thesis
Year:  2011
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Abstract:
Perna viridis¨alias name is Mussels screw,which is widely distributed the south of East China Sea and South China Sea, and its ability to reproduce is very strong and easy to breed.But due to a lack of market and effective processing method, its resources failed to be fully utilized. Atherosclerosis、fatty liver、heart cerebrovascular disease has become the major disease which is harmful to human health, and lipid metabolism disorder, especially hypercholesterolemia is the basic susceptibility factor. Therefore, how to effectively regul- ate blood-fat is currently one of the hot spot of nutritional research.Systemic research on the isolation and purification of Perna viridis from glycosa- minoglycan was studied in this paper,then analyzing and determining its physicoc- emical property.The effects of Perna viridis from glycosaminoglycan on the lipid metabolism were detemined. At the same time,investigation on the effects of Perna viridis from glycosaminoglycan on the anti-oxidation in hyperlipidemia mice was conducted.The resu- lts can provide theoretical basis for evaluating Perna viridis from glycosaminoglycan nutr- ition value and the development of Perna viridis for reference It can have a contribution to the development of shellfish.The main results are as follows:(1) Glycosaminoglycan Perna viridis was largely extracted on the optimal conditions, the removal of protein in the product was compared by different pH values,and then mem- brane filtration and desalination was used to prepare PVG-1.PVG-1was purified by DEAE -52 column chromatography, higher content of PVG-1-1,PVG-1-2 were obtained,further purified by Sephadex G-200 chromatography,PVG-1-1a,PVG-1-3a were obtained. The chemical composition of PVG、PVG-1、PVG-1-1、PVG-1-3、PVG-1-1a、PVG-1-3a were examed. The total sugar of PVG、PVG-1、PVG-1-1、PVG-1-2、PVG-1-1a、PVG-1-3a is respectively 20.34%、26.51%、40.64%、37.56%、49.56%、43.12%;62.1%;the content of GAG is respectively 26.81%、45.36%、66.78%、60.45%、75.56%、69.78%;the content of protein is respectively 33.43%、18.12%、13.85%、11.89%、10.75%、8.45%;the content of sulfate is respectively 4.53%、6.89%、12.47%、14.85%、19.65%、21.56%; the content of galactosamine is respectively 2.32%、3.71%、6.64%、8.65%、7.67%、10.56%; the content of uronic acid is respectively 5.88%、7.89%、12.78%、10.67%、19.85%、16.56%;the content of total hexosamine is respectively 10.21%、19.36%、29.61%、33.42%、35.67%、39.67%, PVG-1-1a、PVG-1-3a was tested by HPLC for purity and the result showed that PVG-1-1a and PVG-1-3a both had a characteristic absorption peak, they are homogeneous polysac- charide.Relative molecular mass of PVG-1-1a and PVG-1-3a is respectively 5.20〜10~5Da and 2.58〜10~5Da;Infrared spectroscopy analysis their construction and characteristic groups, the results showed that PVG-1-1a and PVG-1-3a both have characteristic absorption peak of glycosaminoglycan.(2)72 kunming male mice were divided into normal contrast group,hyperlipidemia contrast group,positive contrast group,low dose group,medium dose group,and high dose group respectively.The normal contrast group was fed with normal diet and other five groups with high-lipid diet.The normal contrast group and the model contrast group were fed with the same volume of NS,the positive group with Simvastatin Tablets at the dosages of 5 mg/kg,The low dose group,medium dose group and high dose group were fed with glycosaminoglycan from Perna viridis at the dosages of 50mg/kg,100mg/kg and 200mg/kg respectively.The contents of total cholesterol(TC),triglyceride(TG),low density lipoprotein- cholesterol(LDL-C),high density lipoprotein-cholesterol (HDL-C)and NO in serum,the contents of total cholesterol (TC),triglyceride (TG) in liver,the activities of lecithin chole- sterol acyltransferase(LCAT) in serum,lipoprotein lipase(LPL) and hepatic lipase( HL) in serum and liver were examined on the 30th day.Results showed that glycosaminoglycan from Perna viridis had no effect on mice weight (P0.05),the middle-dose and high-dose groups can increase liver index(P0.05),decrease renal index (P0.05).The middle-dose and high-dose groups of glycosaminoglycan from Perna viridis could decrease the contents in serum TC,TG,LDL-C,AI(P0.01) and increase serum HDL-C、NO(P0.01) significantly, decrease the contents in liver TC、TG significantly(P0.01),enhance the activities of serum LCAT,serum and liver HPL、HL (P0.05 or P0.01) markedly. Glycosaminoglycan from Perna viridis obviously decreased the blood lipids.The function may be related to improve the activities of LCAT and LPL and increase the level of HDL-C、NO.(3)PVG-1 antioxidant studies in vitro:compared with vitamin C,we test PVG-1 oxidation resistance activeness through the,OH、DPPH,、O2-,and resistance of liver lipid peroxidation in vitro.when PVG-1 concentration is 8mg/mL,,OH clearance rate could reach 55.17%,at the same concentration of PVG,,OH clearance rate could reach 44.59%. The results of antioxidation test in vtro showed that PVG and PVG-1 able to scavenge DPPH,in the concentration of 8.0mg/mL and the scavenging rate was respectively 45.09% and 61.96%.PVG-1 was obviously able to scavenge super oxide free radical in the concent- ration of 5.0mg/mL and the scavenging rate was 68.36%,PVG could scavenge super oxide free radical in the concentration of 6.0mg/mL and the scavenging rate was 56.98%;while PVG and PVG-1 in the concentration of 7.0 mg/mL,the inhibition rate of liver lipid peroxidation respectively reach 46.78% and 62.57%.Experiments in vitro showed that glycosaminoglycan from Perna viridis played a good role in the antioxidation process,this inhibition has a dose-effect relationship.Investigation on the effect of PVG-1 on the antioxidase in hyperlipemia mice was conducted through examining the indexes followed.Total antioxidant capacity in serum and liver,the content of MDA,the activities of GSH-Px、CAT and SOD in serum、liver and brain were determined. The results showed that middle dose group and high dose group of PVG-1 could improve T-AOC(P0.05 or P0.01),PVG-1 could reduce the content of MDA (P0.01)both of serum、liver and brain,increase the activity of GSH-Px(P0.05 or P0.01)、CAT(P0.01)、SOD(P0.01) of serum;the activities of GSH-Px(P0.01)、CAT(P0.05 or P0.01)、SOD (P0.01)and the activities of GSH-Px(P0.01)、CAT(P0.01)、SOD(P0.05 or P0.01) of brain.It can be concluded that PVG-1 could promote the activities of endogenous anti-oxidation enzymes and reduce the level of peroxidation in body,which attribute to the effect of preventing AS.
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