| As the highest economically valuable bamboo species,Moso bamboo(Phylbstachys pubescens)is widely planted in china,with an annual production of about 7 billion roots.Despite the considerable presence of various beneficial active components,the leaves of Moso bamboo were generally discarded without effective utilization,which cause a huge waste of forestry resource.This research can enrich the resource reservoir of natural products and offer valuable reference to the development of activated component In this research,we firstly extracted the active crude polysaccharide from Moso bamboo leaves(CPMBL)using enzymic methods.And the isolation and purification technology was systematically studied,as well as the chemical composition and the biological activities of PMBL.CPMBL was isolated by combined membrane technology and purified by DEAE-cellulose column chromatography;The bacteriostatic and antimicrobial effects of PMBL were observed by plate way.The antioxidant effects in vitro of PMBL were invested by studying the scavenging capacities of ROS,1,1-diphenyl 2-picrylhydrazyl(DPPH) and the antioxidant activities on lipids(TBA),etal.Finally,we investigated several biological functions of PMBL by animal experiment,including bacteriostasis,anti-oxidation,anti-fatigation and decreasing blood sugar and blood fat,respectively.Research results and analysis1.Isolation,purification and composition analysis of PMBLCPMBL was purified and refined PMBL was obtained,further research showed that:two different components were obtained by combined membrane isolation,including ultrafiltrationing polysaccharide solution(UPMBL,MW>6000D)and nanofiltration polysaccharide solution(NMBL,2000DW<6000D),Its polysaccharide purity is 37.37%and 51.76%respectively.In UV detection proved that there is no obvious characteristic absorption peaks for either protein or nucleic acid.Seven pure PMBL obtained from UPMBL and NPMBL by DEAE-cellulose column chromatography.with three relatively higher production among them,Its molecular weight and purity were assayed by HPLC.Result show that they can be proved to be composed with high purity polysaccharides,and its molecular weight were UPMBLW(20378D),UPMBL1(14367D),NPMBLW(5932D)respectively.which are mainly rhamnose,arabinose,mannose,galactose,glucose and fructose as well.Analysis of infrared spectrum showed that,there is little difference in sugar residue composition between UPMBLW and NPMBLW.Experimental results proved that above methods are feasible to isolate and purify the CPMBL in order to get pure PMBL production.2.The antimicrobial activity effect of PMBLThe bacteriostatic and antimicrobial effect of CPMBL and PMBL on gram-positive bacteria(G+)and gram-negative bacteria(G-)were observed.The Mowings are the results: CPMBL has strong bacteriostatic and antimicrobial effects,but PMBL has no obvious antimicrobial activity; Result show that CPMBL has broad - spectrum antimicrobial.and it hasn't effects on fungi. PMBL hasn't bacteriostatic and antimicrobial effects3,The antioxidant effect in vitro of PMBLThe antioxidant effects in vitro of PMBL were invested by studying the scavenging capacities of ROS,1,1-diphenyl 2-picrylhydrazyl(DPPH)and the antioxidant activities on lipids using thiobarbituric acid(TBA)method.The followings are the results:the scavenging effect of PMBL on ROS is good,on O2<sup>IC50=5.98mg/Land·OH(IC50=5.55mg/L)are stronger than Vc's(IC50=6.59mg/L)and rutin's(IC50=17.77mg/L.on DPPH free radical,the scavenging effect of PMBL(IC50=36.08mg/L)is good,but weaker than Vc's(IC50=21.05mg/L)and rutin's (IC50=15.88mg).PMBL has some antioxidant activities on linoleic acidThe erythrocyte hemolytic test results show that,PMBL has obvious inhibition to erythrocyte hemolysis caused by H2O2,the inhibition rate of 30mg/l PMBL is 38.119%,stronger than Vc's and rutin's.and With the concentration increasing,the inhibition is more obvious,which shows that there are obvious dose-effect relationship for PMBL to erythrocyte hemolytic caused by H2O2. The results showed that:The antioxidant effects in vitro of PMBL embody mainly that it has significant scavenging effect on oxygen free radical,and has more weak scavenging effect on organic free radical including DPPH,lipid peroxidation et.al;The antioxidant ability in vitro of PMBL has a dose-effect relationship obviously.4.The antioxidant effect of PMBL in vivo on aged miceCompared with normal aged mice,the antioxidant effect in vivo of PMBL on aged mice was studied.The followings are the results:PMBL can significantly increase contents of protein in the serum,liver & kidney,decrease the contents of MDA in the serum and the organs, increase activity of SOD and GSH-Px in the serum,heart,liver,kidney and the total antioxidant competency.The ratio of TAOC/MDA shows that,TAOC/MDA of PMBL group in the serum, heart,liver,kidney is obvious higher than the normal group.The results show that,PMBL can promote portein synthesis in serum and organs,mitigate recession of the tissue and cytoarchitecture,improve the function,balance the state of antioxidation/oxidation.5.antifatigue effects of PMBLCompared with normal mice,antifatigue eifects of PMBL was studied by intragastric infusion different dose of PMBL.The followings are the results:PMBL could extend swimming time,decrease the content of blood urea nitrogen,increase the activity of Lactic dehydrogenases,obviously increase reservation of liver glycogen and muscle glycogen.The results show that PMBL has physical fatigue alleviating function6,The hypoglycemic effect of PMBLMice were injected alloxan to cause diabetic model,and the hypoglycemic effect of PMBL on experimental diabetic mice and the influence on the serum glucose of normal mice were observed respectively.The following are the results:PMBL could remarkably decrease the serum glucose,relate weight lose and water intake of diatetic mice,but has no harmful effects on the serum glucose of normal mice.The results show that PMBL has hypoglycemic effect significantly on diatetic mice,and has no harmful effects on normal mice.7,Regulation of PMBL on lipid metabolism(1)Preventive effects of PMBL on hyperlipidemia and fatty liver were evaluated by ig. different dosages PMBL to the mice fed with high fat diet,the experiment lasted for 4 weeks. The followings are the results:PMBL has obvious inhibition to weight gain of the mice caused by high fat feeding,and good relief function to growth disorder caused by liver damage.PMBL can remarkably inhibit the rising of blood lipid,effectively decrease TC,TG,LDL-C in the serum,increase HDL-C in the serum.All dose of PMBL can reduce the liver cell damage, decrease ALT activity in the serum and the content of liver crude fat,inhibit the liver index and hepatomegaly,and the effect is batter than positive control(PC,Xuezhikang).The results show that PMBL can effectively relief and prevent the formation of hyperlipidemia,fatty liver and liver damages.(2)The lipid-reducing and liver-benefiting effects of PMBL on the hyperlipidemia,fatty liver and liver-damaging mice were studied.The experiment lasted for 2 weeks.The followings are the results.Different dose PMBL all can remarkably decrease TCHO,TG,LDL-C contents in the serum,and improved HDL-C contents in the serum appreciably;Reduce the content of liver crude fat(the highest reduction rate are 58%),the liver function and liver cell morphometry in PMBL group are superior to HL group significantly,and similar to NG.The study conformed that,All the dose PMBL can remarkably decrease the content of liver crude fat,ALT activity and liver index in the with hyperlipidemia,fatty liver and liver damages.The results show that PMBL has remarkable antilipidemic effect,and can effectively improve the liver function on the mice with hyperlipidemia and fatty liver.liver damage.The research results show that,the application of membrane separation technology is simple and feasible,which can provides a reference to the study of other plant polysaccharides.PMBL has activity of antifatigue,antioxidative,hypoglycemic and antilipidemic in certain degree, which shows that non-traditional medicinal plant polysaccharides has the same biological activities with medicinal plant polysaccharides,non-traditional medicinal edible resource can be used to product polysaccharides to improve the deficient state of excessive exploitation of Chinese Herbal Drugs Resources,and helpful to improve the added value of moso bamboo leaves, promote the development of forestry economy provide new methods and models for related resources...
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