【摘要】 目的探讨促红细胞生成素(EPO)对慢性环孢素A(CsA)肾毒性致细胞凋亡的抑制作用及机制。方法 SD大鼠分为四组:对照组:给予橄榄油(1 mg·kg-1·d-1皮下注射);对照组+EPO(100 U/kg,3次/w腹腔注射);CsA毒性组:给予CsA(15 mg·kg-1·d-1皮下注射);CsA+EPO。检测各组大鼠的体重、肾功能、收缩期血压、血色素水平;肾小管间质纤维化和细胞凋亡分别以三色染色和TUNEL染色观察;利用RNA原位杂交和免疫印迹法检测转化生长因子(TGF)β1诱导基因h3(βig-h3)、细胞凋亡相关基因的表达。结果与对照组相比,CsA毒性组体重下降、肾功能低下、贫血,伴有肾小管间质纤维化和大量TUNEL阳性细胞。在分子水平上,CsA肾毒性组βig-h3 mRNA和蛋白的表达增加;凋亡基因Fas和Bax表达上调,而Bcl-2蛋白表达减少。EPO治疗均使上述指标逆转(P<0.05)。直线相关分析显示,肾小管间质纤维化程度与βig-h3蛋白表达(r=0.835,P<0.001)和TUNEL阳性细胞数呈正向相关(r=0.698,P<0.001)。结论在慢性CsA肾毒性中,EPO抑制细胞凋亡具有抗纤维化的肾脏保护作用。更多还原

【Abstract】 Objective To examine the anti-apoptotic effect of erythropoietin( EPO) in a rat model of chronic cyclosporine A( CsA) nephrotoxicity. Methods Sprague-Dawley rats were divided into Vehicle( VH),VH+EPO,CsA,CsA+EPO groups. Renal function,systolic blood pressure,and hematopoietic parameters were measured. In addition,renal histopathology( TIF),apoptotic cell death( TUNEL assay),and the expressions of transforming growth factor( TGF) β1 inducible gene-h3( βig-h3) and apoptosis-related genes were evaluated by in situ hybridization and immunoblotting. Results Compared with the VH-treated rats,the CsA-treated rats showed loss of body weight, renal dysfunction,and anemia,with the development of tubulointerstitial fibrosis and increased TUNEL-positive cells. In a molecular level, CsA treatment induced a significant increase in the expressions of βig-h3,Fas,and Bax,whereas Bcl-2 expression was decreased significantly by CsA. Concomitant treatment of EPO reversed all of above parameters. Of note,the degree of the tubulointerstitial fibrosis correlated with expression of βig-h3 and the number of TUNEL-positive cells. Conclusions EPO effectively abrogates the tubulointerstitial fibrosis by inhibiting apoptotic cell death in a rat model of chronic CsA-induced nephrotoxicity.更多还原