中文摘要：

目的研究糖原合成酶激酶（GSK）3β过表达及GSK3β抑制剂SB-216763通过Wnt通路对大鼠肝卵圆细胞增殖的影响及其调控机制。方法肝卵圆细胞WBF-344分为空白对照组、GSK3β过表达慢病毒组（过表达组）、二甲基亚砜对照组和GSK3β抑制剂组，抑制剂组设1、5、10μmol／L3个浓度梯度。以鉴定正确的GSK3β过表达慢病毒和不同浓度SB-216763处理WBF-344细胞，相差及荧光显微镜分别观察细胞形态及慢病毒组细胞荧光表达量；CCK8检测细胞增殖，AnnexinV-碘化丙啶检测细胞凋亡；Western blot法检测GSK3β、β—catenin及cyclinD1蛋白表达。结果镜下见GSK3β过表达组细胞较少，老化明显，且出现大量绿色荧光；各抑制剂组细胞分裂旺盛，状态良好，且细胞数随SB-216763浓度升高而增多。CCK8及流式细胞术显示GSK3β过表达组细胞增殖减慢（t=7．178，P〈0．01），凋亡明显，抑制剂组增殖加快（F=45．030，P〈0．01）。Westemblot法显示过表达组GSK3β呈高表达趋势，p—catenin和cyclinD1表达减弱，抑制剂组GSK3β表达量无明显差别，但随抑制剂浓度增高，B—catenin和cyclinD1表达增强。结论GSK3β过表达可下调Wnt通路使细胞增殖减慢，促进凋亡。GSK3β抑制剂能激活Wnt通路促进细胞增殖。

英文摘要：

Objective To research the effects of glycogen synthase kinase （GSK3β） overexpression and GSK3β inhibitor SB-216763 on the proliferation of hepatic oval cells in rats and its regulatory mechanisms by Wnt signaling pathway. Methods The hepatic oval cells WBF-344 were divided into the blank control group, GSK3 β over-expression group, DMSO control group and GSK3 β inhibitor groups, while the inhibitor groups set up three concentration gradients, that was 1, 5, 10 μmol/L. Using the GSK3β overexpression lentivirus, which had been identified correctly, and SB-216763 dealt with the cells WBF-344. The cells morphology of each group was observed under the phase contrast inverted microscope, and the expression of fluorescence in the lentivirus-transfected group was observed under the fluorescent microscope. The proliferation of each group cells was tested by CCK8 kits. The cells＇ apoptosis was detected by AnnexinV-FITC/PI kits. The expression of GSK3β, β-catenin and cyclin D1 were detected by Western blot. Results The cells of GSK3β over-expression group were fewer and obvious aging. However, in each inhibitor added group, the cells＇ division and proliferation was vigorous, and the condition was good. Moreover, the cells＇ proliferation was getting stronger with the concentration of SB-216763 increasing. A large number of green fluorescence was expressed in the lentivirus-transfected cells. The ceils＇ proliferation in GSK3β over-expression group restrained （t = 7. 178, P 〈 0. 01, as compared with control） , while the cells＇ proliferation was vigorous in inhibitor groups （ F = 45. 030, P 〈 0. 01, as compared with control）. Flow Cytometry showed that the cells apoptosis was significant in GSK3β over-expression group. Western blotshowed that the expression of GSK3β was increased, while the expression of β-catenin and cyclin D1 was decreased in the over-expression group. The expression of GSK3β had no significant difference among the control group and inhibitor groups. However, the expres