生血宁片通过促进内源性EPO生成改善肾性贫血的作用机制研究

As an oral iron agent, Shengxuening tablets has a good effect on improving renal anemia caused by chronic kidney disease in clinic, but its mechanism is still unclear. In our previous study, we found that Shengxuening tablets can increase the hemoglobin and serum EPO level of renal anemia rats, reduce its serum creatinine and urea nitrogen, reflecting that Shengxuening tablets can effectively improve renal anemia. However, the effects of Shengxuening tablets and its underlying mechanisms in improving renal anemia have not yet been investigated. Therefore, we proposed the hypothesis that Shengxuening tablets can supply iron, increase the expression of HIF-2α, promote the secretion of endogenous EPO, then improve renal anemia. In this study, the regulation effect of Shengxuening tablets on EPO expression was clarified from the molecular, cellular, tissue and animal levels. We aim to investigate the regulatory effect of Shengxuening tablets on the binding affinity between IRP1 and HIF-2α mRNA, the effect of Shengxuening tablets on HIF-2α activity and the expression of GATA2 and NF-κB, and to reveal the mechanism of Shengxuening tablets in improving renal anemia by promoting endogenous EPO secretion. This study will open up a new research perspective for the prevention and treatment of renal anemia, and at the same time provide new ideas and approaches for the clinical application of Shengxuening tablets.

生血宁片作为一种口服铁剂，在临床上对慢性肾脏疾病并发的肾性贫血有很好的改善作用，但其作用机制尚不明确。我们前期研究发现，生血宁片的能上调肾性贫血大鼠血红蛋白等血常规指标，降低血清肌酐和尿素氮水平，且生血宁片你能显著增加大鼠血清EPO水平，表明生血宁片能有效改善肾性贫血，但目前未见关于生血宁片防治肾性贫血作用机制的报道。据此我们提出假说：生血宁片能补充铁供应，增加HIF-2α的表达，进而促进内源性EPO的分泌，改善肾性贫血。本课题从分子、细胞、组织以及动物水平等多层次明确生血宁片对EPO表达的调控作用；探讨生血宁片对IRP1与HIF-2α mRNA结合力的调节作用；确定生血宁片对HIF-2α、GATA2、NF-κB表达的调控；揭示生血宁片通过促进内源性EPO分泌这个新视点改善肾性贫血的机制。本研究将为肾性贫血的防治开创了全新的研究视角，同时为生血宁片的临床应用提供新的思路和途径。

生血宁片作为一种口服铁剂，在临床上对慢性肾脏疾病并发的肾性贫血有很好的改善作用，但其作用机制尚不明确。我们通过构建TNF-α诱导HEK-293T细胞炎症模型，研究发现生血宁片在体外能有效抑制GATA2和NF-κB的表达，并显著上调HEK-293T细胞中EPO与HIF-2α的表达。同时，通过0.75%腺嘌呤的饲料喂养大鼠建立大鼠肾贫血模型。ELISA试剂盒检测发现生血宁片组血清白介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的表达水平比CKD组显著降低；试剂盒检测发现生血宁片组的血清肌酐（Cr）和尿素氮（Bun）均明显改善。H&E切片结果表明生血宁片给药后均大鼠肾脏炎症和肾衰竭程度均有所减轻。此外，进一步机制研究发现生血宁片能通过促进HIF-2α表达、抑制PHD2生成，阻断NF-κB和GATA2的转录使EPO表达增加，而且还能通过有效阻断BMP6/SMAD4和IL-6/STAT3信号通路、下调TFR2表达来抑制hepcidin表达。以上结果表明生血宁片可通过促进EPO合成，同时抑制hepcidin调节铁代谢，从而改善大鼠肾性贫血。本研究为肾性贫血的防治开创了全新的研究视角，同时为生血宁片的临床应用提供新的思路和途径。