Small interfering RNA targeting Fas protects mice against renal ischemia-reperfusion injury
Small interfering RNA targeting Fas protects mice against renal ischemia-reperfusion injury
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DOI：
10.1073/pnas.0406421101
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发表时间：
2004-10-12
影响因子：
11.1
通讯作者：
Lieberman, J
中科院分区：
文献类型：
--
作者：
Hamar, P;Song, EW;Lieberman, J
Fas介导的细胞凋亡被认为是肾缺血再灌注损伤后肾小管细胞死亡的原因之一。在这里，我们研究了针对Fas的小干扰RNA(SiRNA)双链是否能保护小鼠在肾动脉夹闭后免受急性肾功能衰竭的影响。夹闭肾静脉和动脉15~35min造成肾缺血再灌注损伤。小鼠在缺血之前或之后接受针对Fas或对照基因的siRNA治疗，通过流体动力注射或低容量肾静脉注射，或两者兼而有之。用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)检测肾组织Fas蛋白和mRNA的表达、组织病理学和细胞凋亡。对摘除对侧肾的小鼠的血尿素氮和存活率进行了监测。单次流体动力注射Fas siRNA可使肾脏Fas基因和蛋白的表达减少4倍。两天前接受Fas siRNA治疗的小鼠的肾脏，TUNEL染色显示肾小管上皮细胞凋亡明显减少，肾小管萎缩和透明层损伤也较少。而用生理盐水或GFP siRNA预处理的小鼠有90%死亡，而Fas-siRNA预处理的动物只有20%死亡。肾静脉注射一次低容量的Fas siRNA也提供了同样的生存优势。此外，通过肾静脉的缺血后注射保护了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。通过全身或局部插管沉默Fas有望限制缺血再灌注损伤。
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.