促红细胞生成素促进内皮祖细胞增殖的体外研究及机制探讨

Effects and possible mechanism of recombinant human erythropoietin on proliferation of endothelial progenitor cells

目的:研究人重组促红细胞生成素(rhEPO)对人内皮祖细胞(EPCs)数量和功能的影响并探讨其作用机制。方法:以不同浓度rhEPO分别作用于15例处于肾衰竭期的患者以及15例健康志愿者EPCs,MTT法检测其增殖,Annexin-V法检测其凋亡,Western blotting法检测Akt蛋白激酶。结果:100U/L、600U/L及1200U/L rhEPO均能提高实验组和对照组EPCs的数量及增殖能力,呈剂量依赖性,实验组EPCs数量及功能均显著低于对照组。1200U/L rhEPO能显著降低EPCs的凋亡率,提高Akt蛋白激酶的表达。加入Wortmannin后能阻断这一效应。结论:rhEPO可以显著提高健康人群和肾衰竭患者EPCs的数量与功能,且这一效应与PI3K/Akt信号通路有关。

AIM： To observe the effects of recombinant human erythropoietin on proliferation of endothelial progenitor cells （EPCs） from healthy volunteers and patients with renal failure, and tried to elucidate the possible mechanism. METHODS： Various concentrations of rhEPO were added to the culture system of EPCs from 15 cases of patients with renal failure （ RF group） and 15 cases of healthy volunteers （ control group）. MTT assays were used to detect proliferative rates. Annexin - V/PI stains were used to measure the apoptotic rates. Western blotting was used to determine the expression of Akt protein kinase. RESULTS： Numbers and proliferative ability of EPCs from control group and RF group were improved in dose - dependent manner when concentrations of rhEPO were 100 U/L, 600 U/L and 1 200 U/L. However, compared to the control group, numbers and function of EPCs from RF group were remarkably decreased. The apoptosis rate of EPCs was decreased and the activity of Akt protein kinase was improved in the presence of 1 200 U/L rhEPO. Wortmannin was able to block the effects. CONCLUSION： rhEPO improves the number and function of EPCs from both healthy volunteer and patients with renal failure. PI3K/Akt might play an important role in it.