Hamar Péter, Song Erwei, Kökény Gabor, Chen Allen, Ouyang Nengtai, Lieberman Judy
Institute of Pathophysiology, Semmelweis University, Nagyvárad tér 4, Budapest, H 1089, Hungary.
Proc Natl Acad Sci U S A. 2004 Oct 12;101(41):14883-8. doi: 10.1073/pnas.0406421101. Epub 2004 Oct 4.
Fas-mediated apoptosis has been suggested to contribute to tubular cell death after renal ischemia-reperfusion injury. Here we investigate whether small interfering RNA (siRNA) duplexes targeting Fas protect mice from acute renal failure after clamping of the renal artery. Renal ischemia-reperfusion injury was induced by clamping the renal vein and artery for 15 or 35 min. Mice were treated before or after ischemia with siRNA targeting Fas or a control gene, administered by hydrodynamic injection, low-volume renal vein injection, or both. Treated mice were evaluated for renal Fas protein and mRNA expression, tissue histopathology, and apoptosis by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) staining. Blood urea nitrogen and survival were monitored in mice in which the contralateral kidney had been removed. A single hydrodynamic injection of Fas siRNA reduced Fas mRNA and protein expression in the kidney 4-fold. Kidneys from mice that received Fas siRNA two days earlier had substantially less renal tubular apoptosis by TUNEL staining and less tubular atrophy and hyaline damage. Whereas 90% of mice pretreated with saline or GFP siRNA died, only 20% of Fas-siRNA-pretreated animals died. The same survival advantage was provided by a single low-volume Fas siRNA injection into the renal vein. Moreover, postischemic injection through the renal vein protected 38% of mice from death. This study confirms the importance of Fas-mediated apoptosis in renal ischemia-reperfusion injury. Silencing Fas by systemic or local catheterization holds therapeutic promise to limit ischemia-reperfusion injury.
Fas介导的细胞凋亡被认为与肾缺血再灌注损伤后的肾小管细胞死亡有关。在此，我们研究靶向Fas的小干扰RNA（siRNA）双链体是否能保护小鼠免受肾动脉夹闭后的急性肾衰竭。通过夹闭肾静脉和动脉15或35分钟诱导肾缺血再灌注损伤。在缺血前或缺血后，通过流体动力学注射、低容量肾静脉注射或两者联合的方式，用靶向Fas或对照基因的siRNA处理小鼠。对处理后的小鼠进行肾Fas蛋白和mRNA表达、组织病理学以及通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）染色检测细胞凋亡的评估。监测对侧肾脏已被切除的小鼠的血尿素氮水平和存活率。单次流体动力学注射Fas siRNA可使肾脏中的Fas mRNA和蛋白表达降低4倍。通过TUNEL染色，两天前接受Fas siRNA处理的小鼠肾脏中的肾小管凋亡明显减少，肾小管萎缩和透明变性损伤也减少。用生理盐水或绿色荧光蛋白（GFP）siRNA预处理的小鼠中有90%死亡，而用Fas siRNA预处理的动物中只有20%死亡。单次低容量肾静脉注射Fas siRNA也能提供相同的存活优势。此外，缺血后通过肾静脉注射可使38%的小鼠免于死亡。本研究证实了Fas介导的细胞凋亡在肾缺血再灌注损伤中的重要性。通过全身或局部导管插入术沉默Fas有望限制缺血再灌注损伤。
