Darbepoetin alfa protects podocytes from apoptosis in vitro and in vivo
Darbepoetin alfa protects podocytes from apoptosis in vitro and in vivo
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DOI：
10.1038/sj.ki.5002362
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发表时间：
2007-08-01
影响因子：
19.6
通讯作者：
Shankland, S. J.
中科院分区：
文献类型：
--
作者：
Logar, C. M.;Brinkkoetter, P. T.;Shankland, S. J.
足细胞的脱落或凋亡导致蛋白尿和肾小球硬化。目前还没有专门预防足细胞凋亡的糖尿病或非糖尿病肾小球疾病干预措施。红细胞生成刺激蛋白（ESP）与非造血细胞上的受体的结合已被证明在体外、体内和初步的人体研究中具有抗凋亡作用。最近，在足细胞上鉴定了促红细胞生成素受体;因此，我们检测了达贝泊苷α预防足细胞凋亡的作用。培养的永生化小鼠足细胞用低剂量紫外线-C（uv-C）照射，诱导细胞凋亡的存在或不存在的达贝泊苷α。通过Hoechst染色和通过Western印迹评估的半胱天冬酶3切割来定量细胞凋亡。用达贝泊汀α预处理显著减少足细胞凋亡，这种作用涉及完整的Janus家族蛋白激酶-2（JAK 2）和AKT信号通路。此外，发现达贝泊苷α对转化生长因子-β 1具有保护作用，但对嘌呤霉素氨基糖苷诱导的细胞凋亡无保护作用。当用达贝泊汀α治疗时，抗肾小球抗体诱导的肾小球肾炎小鼠的蛋白尿、肾小球硬化和足细胞凋亡显著减少。我们的研究表明，用ESP治疗以足细胞凋亡为特征的进行性肾脏疾病可能有利于减缓慢性肾脏疾病的进展。
Detachment or apoptosis of podocytes leads to proteinuria and glomerulosclerosis. There are no current interventions for diabetic or non-diabetic glomerular diseases specifically preventing podocyte apoptosis. Binding of erythropoiesis stimulating proteins (ESPs) to receptors on non-hematopoietic cells has been shown to have anti-apoptotic effects in vitro, in vivo, and in preliminary human studies. Recently, erythropoietin receptors were identified on podocytes; therefore, we tested effects of darbepoetin alfa in preventing podocyte apoptosis. Cultured immortalized mouse podocytes were treated with low-dose ultraviolet-C (uv-C) irradiation to induce apoptosis in the absence or presence of darbepoetin alfa. Apoptosis was quantified by Hoechst staining and by caspase 3 cleavage assessed by Western blots. Pretreatment with darbepoetin alfa significantly reduced podocyte apoptosis with this effect involving intact Janus family protein kinase-2 (JAK2) and AKT signaling pathways. Additionally, darbepoetin alfa was found protective against transforming growth factor-beta 1 but not puromycin aminonucleoside induced apoptosis. Mice with anti-glomerular antibody induced glomerulonephritis had significantly less proteinuria, glomerulosclerosis, and podocyte apoptosis when treated with darbepoetin alfa. Our studies show that treatment of progressive renal diseases characterized by podocyte apoptosis with ESPs may be beneficial in slowing progression of chronic kidney disease.