PIM-3激酶通过与AQP2作用调节肾脏水代谢的机制研究

PIM-3 Kinase Regulating Kidney Water Metabolism by Interacting with AQP2

目的:研究HK-2细胞中PIM-3激酶与AQP2相互作用调节肾脏水代谢的作用机制。方法:免疫荧光检测PIM-3激酶表达水平;过表达和沉默PIM-3激酶,观察细胞增殖、凋亡及迁移率变化;Western blot检测AQP2、p-AQP2以及细胞凋亡因子表达水平。结果:PIM-3激酶在HK-2细胞中大量表达;PIM-3激酶过表达后,HK-2细胞增殖能力和迁移率提高(P<0.01),细胞凋亡率降低(P<0.05);同时伴随p-AQP2表达明显上调(P<0.01),而促凋亡因子Bax、Cleaved Caspase-3表达明显下降(P<0.05);沉默PIM-3激酶结果完全相反。结论:PIM-3激酶在HK-2细胞中表达,其可能通过促进AQP2蛋白的磷酸化影响HK-2细胞的活性、迁移等生物学特征,从而改善肾脏水代谢失衡,为肾性水肿的治疗提供新的分子机制和治疗靶点。

Objective:To study the mechanism of interaction between PIM-3 kinase andAQP2 in HK-2 cells to regulate renal water metabolism.Methods:The expression level of PIM-3 kinase in HK-2 cells was detected by immunofluorescence;after over-expression and siRNA gene silencing of PIM-3 kinase in HK-2 cells,changes in cell proliferation,apoptosis and migration rate were observed;the expression levels of AQP2,P-AQP2 and apoptosis factors were detected by Western blot.Results:PIM-3 kinase was highly expressed in HK-2 cells;after PIM-3 kinase was overexpressed,cell proliferation and cell migration rate was enhanced(P<0.01),cell apoptosis rate was reduced(P<0.05);at the same time,p-AQP2 expression was significantly up-regulated(P<0.01),and the expression of pro-apoptotic factors Bax and Cleaved Caspase-3 decreased significantly(P<0.05).After silencing the PIM-3 kinase gene,the results were completely opposite.Conclusion:PIM-3 kinase is expressed in HK-2 cells,and it may affect the biological characteristics of HK-2 cells such as activity and migration through inducing phosphorylation of AQP2 protein,thereby improving the imbalance of renal water metabolism.It will provide new molecular mechanisms and therapeutic targets for the treatment of renal edema.