氯化锂通过激活Wnt信号通路促进内皮祖细胞的增殖能力  

Glycogen synthase kinase 3β inhibitor lithium chloride increased the proliferation of endothelial progenitor cell by activating Wnt signaling pathway

作　　者：崔斌[1] 刘曦[1] 李佳蓓[1] 秦浙学[1] 于世勇[1] 黄岚[1] CUI Bin LIU Xi LI Jia-Bei QIN Zhe-Xue YU Shi-Yong HUANG Lan(Institute of Cardiovascular Disease, Xinqiao Hospital, Third Military Medical University of PLA, Chongqing 400037, Chin)

机构地区：[1]第三军医大学新桥医院全军心血管内科研究所,重庆市400037

出　　处：《中国动脉硬化杂志》2017年第10期985-988,1001,共5页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金资助项目(81100149)

摘　　要：目的探讨糖原合酶激酶3β(GSK-3β)抑制剂氯化锂对内皮祖细胞增殖的作用机制。方法密度梯度离心法分离大鼠骨髓源性内皮祖细胞,体外培养7天后分别给予不同浓度氯化锂(5、10、20、40 mmol/L)以抑制内皮祖细胞的GSK-3β活性。分别采用镜下计数法及四氮唑溴盐比色法测定内皮祖细胞增殖能力,荧光激活细胞分离仪分析各组内皮祖细胞的细胞周期变化。采用蛋白印迹法测定Wnt信号通路中磷酸化GSK-3β、β-连环蛋白及细胞周期蛋白D1的蛋白表达。结果氯化锂在一定浓度范围内可剂量依赖性地增加内皮祖细胞数量。与对照组比较,不同浓度氯化锂组(5、10、20 mmol/L)内皮祖细胞数量显著增加(P<0.05,n=5)。四氮唑溴盐比色法检测氯化锂各浓度组内皮祖细胞在490 nm处吸光度值与对照组比较差异有显著性(P<0.05,n=5)。荧光激活细胞分离仪分析显示氯化锂组细胞周期S期较对照组显著增加(P<0.05,n=3)。蛋白印迹法测定表明,与对照组比较氯化锂可显著增加磷酸化GSK-3β、β-连环蛋白及细胞周期蛋白D1的蛋白表达(P<0.05,n=3)。结论 GSK-3β抑制剂氯化锂可通过激活Wnt信号通路显著促进内皮祖细胞的增殖能力。Aim To investigate the mechanism of glycogen synthase kinase 3β （GSK-3β）inhibitor lithium chlo- ride on the proliferation of endothelial progenitor cells （EPC）. Methods Mononuclear cells were isolated from bone marrow in rats by density gradient centrifugation combined. After 7 days, EPC were cultured with different concentrations of GSK-3β inhibitor lithium chloride. EPC proliferation were assessed by cells count and 3-{4,5-dimethylthiazol-2yl} -2, 5-diphenyltetrazolium bromide （MTT） assay. The cell cycle of EPC was measured by fluorescence-activated cell sorting （FACS）. The expression of phosphor-GSK-3β（ pGSK-3β）, β-catenin and cyclinD1 in EPC were detected by Western blot. Results Lithium chloride improved EPC proliferation. The number of EPC was obviously increased in lithium chloride group than that in control group. Compared with control group, EPC proliferation was enhanced in a dose-depend- ent manner in lithium chloride groups. S phrase in cell cycle was increased in lithium chloride group in comparison with control group by FACS assay. The protein expression of pGSK-3β, β-catenin and cyclinD1 were significantly higher than that in control group. Conclusion GSK-3β inhibitor lithium chloride could enhance EPC proliferation by activating Wnt signal pathway.

关 键 词：糖原合酶激酶3Β 内皮祖细胞 氯化锂 细胞增殖 WNT信号通路 

分 类 号：R363[医药卫生—病理学]