The Active Ingredients Of Alpinae Oxyphyllae To Improve Memory And Urine-reducing Function

Alpinae Oxyphyllae Fructus(AOF)is dehydrated fruit of traditional Alpinae Oxyphyllae Miq,a Chinese medicine.Its bioactivities are various,including memory improvement,warming kidney and urine-reducing,neuroprotection,anti-oxidation and aging.The composition of AOF is complex.Previous studies focus on the chemical constituents and pharmacological activities.However the research about bioactive ingredients,the most important elements in AOF,are rarely reported.In this study,we try our effort to analyze the potential relationship between the AOF main bioactive ingredients and the multiple bioactivity.The research will lay a theoretical foundation on the further development of AOF.(1)The study about memory improvement and urine-reducing activity in different crude component of AOF.After extracted by different solvents,we obtained the ethanol extract(A),the water extract of(B),ethyl acetate extract(C),n-butanol extract(D)and extract residue(E).The mouse water maze test and the platform test were used to explore the effect of each crude component on the memory activity of mice.It was concluded that A can significantly improve the memory of mice(P < 0.01 or P < 0.05).Some of them contain active substances for improving memory;at the same time,the urine volume of mice was collected,it was concluded that C of AOF can quickly inhibit the production of urine in a short period of time(P < 0.01 or P < 0.05),indicating that the component contains a urine-reducing active ingredient.(2)The study about isolation of subdivided components of the alcoholic water extract and activity of memory improvement.The component A was filtrated by different columns,to remove impurity and harvest different components: L,M,and N.The water maze and the platform test were employed to explore the function of different components.The results showed that the component L could significantly improve the memory;the activity of AchE in the hippocampus was significantly weakened,and the activity of ChAT was significantly enhanced(P < 0.01 or P < 0.05);the component L was repeatedly purified by HW-55 F and Sephacryl S-400 gel column to harvest the final product L11.Its structure were identified by NMR and other spectroscopic methods.Compound L11 has classical polysaccharide structure and its main chain is a(1?2)-?-glucose isomer configuration.(3)The study about isolation of subdivided components of Ethyl acetate extract and activity of urine-reducing excretion.The component C was separated by MCI column,and the gradient elution was carried out by different concentrations of ethanol solution,and the components P1,P2,P3 and P4 were sequentially obtained.The urine-reducing murine models was established by metabolic cage.The total urine volume in the first 2 h and 6 h of the mice was calculated by urinary weight/body weight.The results showed that the urine volume/body weight of P3 and P4 groups showed a deceasing trend and the effect of P4 group was more significant(P < 0.01 or P < 0.05),indicating those ingredients have urine-reducing activity.P3 and P4 were mixed and then separated and purified by ODS-A and ODS-D columns.Finally,four compounds(I,II,III and IV)were obtained.Identifying the structure by NMR and other spectral methods They were identified as 3-methoxy-4hydroxy-diphenylhexane,20-propyl-?-sitosterol,oleic acid,and linoleic acid,respectively.We concluded that 3-methoxy-4hydroxy-diphenylhexane and 20-propyl-?-sitosterol in AOF are playing important role in urine-reducing.