OBJECTIVE To study the influence of glycogen synthase kinase3β (GSK3β) over expres?sion in the hippocampus on the antidepressant and anxiolytic effects of total flavoids from Xiaobuxin Tang (XBXT-2). METHODS Adeno-associated virus containing GSK3β(S9A) mutation was microinjected into the hippocampus. After three weeks of recovery, GSK3βand p-GSK3βwere detected by Western blotting, and open field test (OFT) was used to evaluate the locomotor activity. Then, AAV group and GSK3β over expression group were divided into administration group and solvent group, respectively. XBXT-2 (100 mg · kg-1) and solvent were ig administered chronically. After 14 d and 16 d of administra?tion, the tail suspension test (TST) and forced swimming test (FST) were used to investigate the influence of GSK3βover expression on the antidepressant effect of XBXT-2, respectively. After 18 d and 20 d of administration, the elevated plus maze test (EPMT) and staircase test (ST) were used to investigate the influence of GSK3β over expression on the anxiolytic effects of XBXT-2, respectively. RESULTS Western blotting analysis showed that the protein level of GSK3βincreased significantly in GSK3βover expression group (P<0.01) compared with AAV group, but there was no significant difference in p-GSK3β. In OFT, the number of crossings and rearings showed no difference between AAV group and GSK3β over expression group. The results of TST and FST showed that compared with AAV group, the immobility time was significantly reduced in AAV+XBXT-2 group (P<0.05, P<0.01), but compared with GSK3β over expression group, the immobility time showed no difference in GSK3β over expression+XBXT-2 group. In EPMT, compared with AAV group, the percentage of entrances and time into open arms in AAV+XBXT-2 group was significantly increased (P<0.01, P<0.05), but compared with GSK3βover expression group, these indexes showed no difference in GSK3βover expression+XBXT-2 group. In ST, compared with AAV group, the number of rearings was significantly reduced in AAV+XBXT-2 group (P<0.05), but there was no difference between GSK3β over expression+XBXT-2 group and GSK3βover expression group. CONCLUSION GSK3βover expression in the hippocampus can reverse the antidepressant effects of XBXT-2 in the TST and FST, and the anxiolytic effects in the EPM and ST.