Hypoglycemic Effect Of Extracts Fromllex Kudincha C. J. Tseng From Hainan In HFD And STZ-induced Diabetic Mice And Its Mechanism Of Action

Objectives: This study was designed primarily to investigate theHypoglycemic effect of of Extracts from Ilex kudincha C. J. Tsengfrom hainan on Diabetic Mice Induced by high fat diet andstreptozotocin.We tried to identify the main bioactive Fractionsof Ilex kudincha C. J. Tseng, and the mRNA levels of key enzymeswhich control glycome tabolism analysised by RT-PCR. In all, wewanted to provide theoretical basis for a novel food-derivedanti-diabetic agents of Kudingcha.Diabetes is the world’s largest endocrine disease, whichcharacterized by elevated blood glucose production and impairedglucose clearance. However, currently available drugs for type2diabetes have a number of limitations, such as adverse effects andhigh rates of secondary failure. Therefore, it is necessary to lookfor new drugs and interventions that can be used to manage thismetabolic disorder.Ilex kudingcha C. J. Tseng (Aquifoliaceae), is widely consumed as a traditional herbal tea known as ’’KuDingCha’’ in southern China,and it is also used in folk medicine and employed in commercialherbal preparations. Nowadays, there are many reports about itschemical composition and pharmaceutical functions. These reportsshowed that Ilex kudingcha have various biological activities. Ithas been claimed to act as a stimulant to the central nervous system,a diuretic,an antirheumatic, an antiobesity, an antidiabetic, anantihypertensive, and also a treatment for sore throats. However,little information about the mechanism for its effect on diabetesof kudingcha made from llex kudingcha is available. Thus the goalof this study was to investigate the hypoglycemic effect of extractsfromIlex kudincha C. J. Tseng from hainan in HFD and STZ-InducedDiabetic Mice and its mechanism of action.Method:1Diabetic Mice model: The mice received a diet containing(w/w) basic diet (78.8%), egg yolk (10%), lard (10%), cholesterol(1%) and cholate (0.2%) for one month and then was injected i.p.witha single dose of STZ (130mg/kg body weight).2Prepare of the extract from Ilex kudingcha: The lyophilizedpowder of water extract from Ilex kudingcha was extracted with4-fold(w/v)100%MeOH at65℃for2h and the extraction wasrepeated for3times. The combined100%MeOH extract was evaporatedand dried by a freeze dryer to yield fraction1.The residue wasthen extracted with50%MeOH/H2O, using same extraction process,to get fraction2. The residue was used as fraction3.3Animal Experimental Design: All animals were randomly allocatedto one of6different4-week treatments, with8mice per group: C:control group with0.4mL·d-1of distilled water; T: Type2diabeticmodel group with0.4mL·d-1of distilled water; P: Type2diabetic positive group with50mg·Kg-1d-1·metformin;F1: Type2diabetictreated group with15g·Kg-1d-1fraction1; F2: Type2diabetictreated group with15g·Kg-1d-1fraction2; F3: Type2diabetictreated group with15g·Kg-1d-1fraction3. After4weeks of treatment,Blood samples and organs from each animal were collected and storedat-80℃for follow-up work.Results:1Fraction1improved the symptoms of diabetic mice, raiseits spirit, increase its intensity of activity and body weight.2Fraction1and Fraction2could decrease the blood glueose ofdiabetic mice effectively and improved its OGTT. In addition,Fraction1and Fraction2significantly suppressed the increase inblood TC and TG levels. And Fraction1was more effective.3Fraction1enhanced the activity of SOD and decreased the levelof MDA. It indicated that Fraction1can enhance the antioxidantability of diabetic mice.4Pathological alterations in the livers: the liver histopath-ologic changes were markedly improved in fraction1and fraction2treated animals by the evidence of fewer hepatic lipid dropletsand less lipid inflammatory area.5Gene Expression: The cholesterol metabolism related gene CYP7A1and LDLR were both up-regulated significantly in Fraction1andFraction2groups.Moreover,Fraction1ingestion up-regulated theexpression of GCK gene and down-regulated the expression of G6pcgene related to glycometabolism. However, there were no significantdifferences in the relative quantities of hepatic IR mRNA among allgroups.Conclusions: In the present study we found that Fraction1treatmentsignificantly reduced the elevated levels of serum glycaemic and lipids in type2diabetic mice and improved the antioxidant abilityof diabetic mice. We found out the most bioactive fraction wasFraction1, but further investigations are in progress to find outthe concrete active component and elucidate the detailed mechanismof Fraction1on glucose metabolism.