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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 low concentration ouabain on the proliferation of OK cells (posterior mouse renal tubular epithelial cells) under low serum culture by using low serum culture medium to simulate the malnourished state of renal insufficiency. Methods: The OK cells were cultured with low concentration of ouabain (1-30 n M) in 0.2% serum. The effects of ouabain on the proliferation of OK cells were detected by MTT assay and Brdu incorporation assay. The expressions of Akt and ERK1 / 2 Phosphorylation of PI3K / Akt and ERK1 / 2 by LY294002 and PD98059 respectively. To observe the effect of PI3K / Akt and ERK1 / 2 on ouabain-induced OK cell proliferation. RESULTS: Low concentrations of ouabain (1-30 nM) promoted the proliferation of OK cells and upregulated the phosphorylation levels of Akt and ERK1 / 2 in OK cells. Specific inhibition of Akt and ERK1 / 2 activation by LY294002 and PD98059 can inhibit ouabain-induced proliferation. CONCLUSION: Low concentrations of ouabain (1-10 nM) can promote the proliferation of OK cells. The PI3K / Akt and ERK1 / 2 signaling pathways are involved in the regulation of ouabain on OK cells.