Studies On Preparing Techniques Of The Soybean Isoflavone Aglycones And Its Anti-AD Activity

【Objective】To formulate the technology of Soybean Isoflavone Aglycones , and establish the method for the content determination of isoflavone aglycone. Study its protection of PC12 cell against oxidative damage induced by H2O2 and neurotoxicity by Aβon cell level. In order to study mechanism that Soybean Isoflavone Aglycones anti- Alzheimer's disease (AD) and providing proofs for developing new clinical anti-AD medicine ,we observe the effect of the Aglycones on AD model mice,s learning and memory, effect on levating serum anti—oxidative , effect on adjusting cholinergic transmitter in cortexs tissue , relievimg neurotoxicity, damage In hippocamps of the mouse were be obversed.【Methods】(1)T o extracte Soybean Isoflavones from defatted Soybean, then use acid hydrolysis to transformat glucoside of Soybean Isoflavones into Aglycones ,refine Aglycones . Establish an HPLC method for the content determination of isoflavone aglycones.(2)We built PC12 cell impaired model induced by H2O2 and Aβ.To study Soybean Isoflavone Activity Aglycones protection of PC12 cell against oxidative damage induced by H2O2 and neurotoxicity by Aβ, We Observe under the microscope , use MTT assay to evaluate the survival rate and LDH to check the LDH activity. (3)After treating with Soybean Isoflavone Aglycones, Y-maze and step experiments were used to observe the effect of on Alzheimer's disease model mice,s learning and memory.To study the effect on serum anti—oxidative, we determinate SOD,GSH-P activity, MDA content. To study the effect on adjustting cholinergic transmitter in cortexs tissue we determinate AchE activity. To study the effect on relievimg neurotoxicity, we determinateβ-Secretase Activity. damage In hippocamps of the mouse were be obversed too to study protection of the Aglycones.【Results】(1)We get the preparing techniques of Soybean Isoflavone Aglycones and establish the method for the content determination.The contents of three Activity Aglycone are 70.98%, Genistein,Daidzein and Glycitein were 46.59%, 21.215 % and 3.175%.(2)After being treated byH2O2 (150umol/L) and Aβ25-35(20umol/L) ,PC12 cell is strongly damaged . The PC12 cell form is abnormaland the quantity of cell is notably decreased.Compare to the normal control ,the the survival rate obviously reduces(P <0.05) and the LDH activity rise.The form of PC12 cells, which were protected by Soybean Isoflavone Activity Aglycone Component ,is normal ; the the survival rate of cell is near normal control and the LDH activity is obviously lower than damaged group (P <0.05).(3)The memory of Alzheimer's disease model mice was decreased significantly,and was improved by Soybean Isoflavone Aglycones treatment.Meanwhile,serum SOD activity, GSH-Px activity, AchE activity in cortexs tissue andβ-Secretase Activity in hippocampus were increased(P <0.05) by Soybean Isoflavone Aglycones treatment,whereas the contents of serum MDA were decreased(P <0.05) .【Conclution】(1)The preparing technique of Soybean Isoflavone Aglycones from defatted Soybean is stable and can be controlled.(2) Our studies show that Soybean Isoflavone Aglycones can obviously improve the abnormal form of PC12 cell induced by H2O2 and Aβand show a good protection of PC12 cell against oxidative damage induced by H2O2 and neurotoxicity by Aβ.(3)Soy isoflavones can improve learning and memory of Alzheimer's disease model mice, elevate the level of serum anti—oxidative, adjust cholinergic transmitter in cortexs tissue , relieve neurotoxicity in central nerve system.