Screening for the Active Ingredients of Ziziphus jujuba with Neuroprotective Effects and Their Mechanisms of Action / 中国药房

ABSTRACT

OBJECTIVE:To conduct screening for the active ingredients of Ziziphus jujuba with neuroprotective effect and to il-luminate their mechanisms of action preliminarily. METHODS:After neuron-like cells (PC12 cells) were respectively cultured in the ingredient of Z. jujuba with polysaccharide enriched(1 mg/ml),that with polysaccharide removed(1 mg/ml),7 kinds of flavo-noid ingredients of Z. jujuba(catechin,procyanidine B2,epicatechin,hyperoside,rutin,quercetin-3-β-D-glucoside and kaempfer-ol-3-O-rutinoside,represented by A,B,C,D,E,F,G,all at the concentrations of 3,13,30 μmol/L)and 2 kinds of nucleoside ingredients of Z. jujuba (cyclic adenosine monophosphate and cyclic guanosine monophosphate,both at the concentrations of 3, 13,30 μmol/L)for 24 h,tert-butyl hydroperoxide(tBHP,150 μmol/L)was used to act on PC12 cells for 3 h to induce oxidative cellular damage,and MTT assay was employed to detect the survival rate of PC12 cells. The PC12 cells transfected with reporter gene plasmid(pARE-Luc)were cultured as above for 24 h,luciferase(Luc)assay was used to detect the transcriptional levels of the antioxidant response element(ARE)of all groups of cells(reflected as the activity of Luc)so as to investigate the anti-injury mechanism. RESULTS:The ingredient of Z. jujuba with polysaccharide enriched could significantly increase the survival rate of PC12 cells to which oxidative damage was caused and the transcriptional level of ARE in the transfected cells. Among the flavonoid ingredients of Z. jujuba, A(30 μmol/L),B(3-30 μmol/L)and C(10-30 μmol/L)could significantly increased the survival rate of the cells,and A(30 μmol/L),B(3-30 μmol/L),C(30 μmol/L),E(30 μmol/L)and F(3-30 μmol/L)could obviously in-creased the activation level of ARE in the transfected cells. However,the nucleoside ingredients of Z. jujuba including cyclic ade-nosine monophosphate and cyclic guanosine monophosphate had no obvious effect of increasing the survival rate of PC12 cells to which oxidative damage was caused or activating the transcription of ARE in the transfected cells. CONCLUSIONS:The polysac-charide and flavonoid ingredients of Z. jujuba may be the active ingredients which account for the neuroprotective effect against oxi-dative cellular damage,and their mechanisms of action may be related to the activation of ARE transcription.