低浓度哇巴因对负鼠肾小管上皮细胞增殖的影响

目的:用低血清培养液来模拟肾脏供血不足的营养不良状态,研究低浓度哇巴因对低血清培养下OK细胞(负鼠肾小管上皮细胞)增殖的影响。方法:用低浓度哇巴因(1-30n M)处理0.2%血清培养下OK细胞,MTT实验和Brdu掺入法检测哇巴因对OK细胞增殖的影响;Western blot检测Akt和ERK1/2的磷酸化水平;用LY294002和PD98059分别抑制PI3K/Akt和ERK1/2蛋白激酶活性,观察抑制PI3K/Akt和ERK1/2对哇巴因促进OK细胞增殖的影响。结果:低浓度哇巴因(1-30n M)促进OK细胞的增值,上调OK细胞中Akt和ERK1/2磷酸化水平。用LY294002和PD98059特异抑制Akt和ERK1/2的活化能够抑制哇巴因的促增殖作用。结论:低浓度哇巴因(1-10n M)能够促进OK细胞的增值,PI3K/Akt和ERK1/2信号通路参与哇巴因对OK细胞促增殖作用的调节。

Low Doses of Ouabain Stimulate Kidney Tubular Cell Proliferation under Serum Deprivation Condition Via Activation of PI3K/Akt and ERK1 /2

Objectivre:To study the effect of physiologic concentration ouabain on OK cell proliferation and to analyze its mechanism.Methods:OK cells cultured under 0.2% Low Serum Culture Medium were treated with 1-30nM ouabain. Brdu incorporation assay and MTT methods were used to detect OK cell proliferation; Western blot was used to determine the phosphorylation of Akt and ERK of OK cells; LY294002, the PI3K/Akt specific inhibitor, and PD98059, the ERK1 /2 specific inhibitor, were used for interfering PI3K/Akt and ERK1 /2 signaling pathway and observed their effect on ouabain induced OK cell proliferationResults:Low dose ouabain (1 -30nM) stimulated OK cell growth. Ouabain could increase the phosphorylation of Akt and ERK in OK cells. The increase of OK cells proliferation by ouabain was inhibited by PI3K inhibitor LY294002 or ERK1 /2 inhibitor PD98059.Conclusion:Low dose ouabain promoted OK cell growth. PI3K/Akt and ERK1 /2 pathway have some effect on regulating this effect.