摘要
目的探讨氯化锂(Li Cl)是否影响近端肾小管上皮细胞(HK-2细胞)的细胞周期及其信号通路。方法将不同浓度(5、12.5、20、25 mmol/L)的Li Cl作用于HK-2细胞不同时间(12、24、48、72 h),采用流式细胞术检测细胞周期变化,CCK-8测定HK-2细胞的活力,Western blotting检测细胞周期G2期的关键蛋白Cyclin B1、CDK1以及AKT/GSK-3β信号通路相关蛋白的表达量。结果流式细胞术结果提示Li Cl以时间-浓度依赖的方式使HK-2细胞的G2细胞周期比例逐渐增加(P<0.05),CCK-8结果提示HK-2细胞的活力随Li Cl作用的时间-浓度的增加而降低(P<0.05),Western blotting显示Cyclin B1、CDK1、p-GSK-3β、β-catenin蛋白含量随着时间以及浓度的增加而增加,而AKT磷酸化水平随着时间及浓度的增加而减少(P<0.05)。结论 Li Cl可能通过激活AKT/GSK-3β信号通路使HK-2细胞阻滞在G2期。
Objective To investigate the effect of lithium chloride（Li Cl） on cell cycle of HK-2 cells and explore the possible pathways involved. Methods HK-2 cells were treated with Li Cl at different concentrations（5, 12.5, 20, and 25 mmol/L） for 12,24, 48, or 72 h, and the changes in cell cycle and viability were detected using flow cytometry and CCK-8 assay, respectively.Western blotting was used to analyze the changes in the expressions of cyclin B1 and CDK1（the two G2 phase-related proteins） and those of AKT/GSK-3β signaling pathway-related proteins in the treated cells. Results Li Cl treatment time-and concentration-dependently increased HK-2 cell percentage in G2 phase and decreased the cell vitality. The expressions of cyclin B1, CDK1, p-GSK-3β, and β-catenin increased and the expression of p-AKT decreased significantly in the cells as Li Cl treatment time and concentration increased. Conclusion Li Cl may cause HK-2 cell cycle arrest in G2 phase through activation of the AKT/GSK-3β signaling pathway.
作者
姜婷婷
张雯英
向晓红
束双双
谢唯
汤珣
章俊
JIANG Tingting;ZHANG Wenying;XIANG Xiaohong;SHU Shuangshuang;XIE Wei;TANG Xun;ZHANG Jun(Department of Nephrology, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2018年第5期541-546,共6页
Journal of Southern Medical University
基金
国家自然科学基金(81202842)
广州市科技计划项目(11C22120703)
广东省科技计划项目(2013B021800149)~~
