Seperation And Identification Of P.koraiensis On PC12Cells Oxidative Damage And Its Physiological Function

Alzheimer’s disease (AD) is an irreversible neurodegenerative brain disease，characterized by progressive loss of memory and other cognitive functions.Evidences indicated that AD have a close relationship with oxidative stress thoughthe pathogenesis of AD is complex. Extensive source of information showed thatphytochemicals isolated from natural products can alleviate oxidative stress in thebrain. Five compounds were isolated from Pinus koraiensis cone by differentchemical solvents, and their inhibition effect on PC12cells were carried out byH2O2induced oxidative damage model. We found that PKBE was the most effectivecompounds on PC12oxidative damage model. Then we seperated the PKBE byAB-8macroporous resin and studied the effect and mechanism of n-butanol extract50%ethanol elution fractions(PKBE-50) on PC12cells induced by H2O2. Finally,we investigated the effects of PKBE on AD model mice. The main contents are asfollows:Pinus koraiensis cone were extracted by95%ethanol, the residue wasextracted by hot water, then the ethanol extracts were extracted by petroleum benzin,formyl trichloride, ethyl acetate and n-butyl alcohol one by one after concentrated,finally determine the chemical components of those extracts. The results showedthat PKBE contains the most polyphenols and flavonoids, while the most abundantcomponents in PKWE is polysaccharide.We found that PKBE have inhibition effect on PC12oxidative damage inducedby H2O2. The PKBE was seperated by AB-8macroporous resin and eluted bydifferent concentration of ethanol, the effect of PKBE elutions were studied byH2O2induced oxidative damage on PC12Cells. Results showed that PKBE-50could improved the viability of the PC12cells significantly and have inhibitioneffect on PC12cells induced by H2O2via decrease the release of LDH, inhibit theproduction of MDA and improve the activity of SOD. PKBE-50could also inhibitthe decrease of MMP and eliminate the ROS in PC12cells.The components constitute of PKBE-50were analysed by LC-ESI-MS. Wefound that PKBE-50were constituted with caffeine acid, amentoflavone,Malvidin-3-O-glucoside, Dihydromyricetin Acetyl glucoside, resveratrol and so on.Finally we assessed the effect of PKBE-50on the learning, memory ability andthe antioxidant enzymes activity of AD mice by injection D-galactose and sodiumnitrite through the abdomina. Results showed that adminstration of PKBE-50couldimprove the learning and memory ability of AD mice, prolong the latency, decrease the error times and reduce the time to found platform when compared to the model.Moreover, PKBE-50could also improve the activity of SOD and GSH-Px in micebrain and decrese the content of lipid peroxide production MDA. In conclusion,PKBK-50have inhibiton effect on oxidative stress.