低浓度哇巴因对负鼠肾小管上皮细胞增殖的影响

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

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。

Objective： 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 1-30 n M 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 proliferation. Results： Low dose ouabain（1-30 n M） 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 PI3 K 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.