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目的:观察重组人促红细胞生成素(recombinant human erythropoietin,rHuEPO)对腹膜间皮细胞(PMCs)的抗凋亡作用及其机制。方法:体外分离小鼠PMCs并给予rHuEPO干预,观察起对JAK2,STAT5,ERK1/2和P38磷酸化的影响。体内试验采用5000U/kg的rHuEPO预处理后再予以4.25%的腹透液腹腔内注射后4小时,取小鼠脏层腹膜组织检测活性凋亡蛋白酶-3表达。结果:小鼠PMCs上表达EPO受体mRNA和蛋白,5U/mL的rHuEPO可诱导PMCs的JAK2,STAT5和ERK1/2磷酸化上调。rHuEPO预处理1小时后再予腹透液处理的PMCs较单独予腹透液处理的PMCs,凋亡蛋白酶-3活化降低,DNA碎裂减少。rHuEPO干预可提高ERK1/2磷酸化水平而抑制腹透液诱导的P38磷酸化。特异性的ERK活化抑制剂PD98059可完全阻断rHuEPO的保护作用。通过检测凋亡蛋白酶-3证实rHuEPO可在小鼠体内降低腹透液引起的PMCs凋亡水平。结论:本研究为rHuEPO在腹透临床治疗中的临床应用开辟了新途径,rHuEPO可通过ERK1/2依赖途径减少PMCs的凋亡发挥保护性作用。rHuEPO及其衍生物可能成为维护腹膜完整性的新治疗手段。
Objective: To investigate the anti-apoptotic effect of recombinant human erythropoietin (rHuEPO) on peritoneal mesothelial cells (PMCs) and its mechanism. Methods: PMCs were isolated from mice and rHuEPO was given to the mice to observe the effect on the phosphorylation of JAK2, STAT5, ERK1 / 2 and P38. In vivo experiments using 5000U / kg of rHuEPO pretreatment and then 4.25% 4 hours after intraperitoneal injection of peritoneal fluid, take the mouse peritoneal layer of peritoneal tissue activity of active caspase-3 expression. Results: EPO receptor mRNA and protein were expressed on PMCs of mice and rHuEPO at 5U / mL could up-regulate the phosphorylation of JAK2, STAT5 and ERK1 / 2 in PMCs. PMCs treated with PDH 1 h after rHuEPO pretreatment showed lower activation of AP-3 and decreased DNA fragmentation compared with PMCs treated with PD alone. rHuEPO intervention can increase ERK1 / 2 phosphorylation and inhibit peritoneal dialysis-induced P38 phosphorylation. PD98059, a specific inhibitor of ERK activation, completely blocked the protective effect of rHuEPO. Pro-apoptotic protease-3 demonstrated that rHuEPO can reduce the level of apoptosis of PMCs induced by peritoneal dialysis in mice. CONCLUSION: This study opens up new avenues for the clinical application of rHuEPO in the clinical treatment of peritoneal dialysis. RHuEPO exerts a protective effect by reducing the apoptosis of PMCs through the ERK1 / 2-dependent pathway. rHuEPO and its derivatives may become new treatments to maintain peritoneal integrity.