Li X, Zhang J F, Lu M Q, Yang Y, Xu C, Li H, Wang G S, Cai C J, Chen G H
Department of Liver Transplantation, The Third Affiliated Hospital of Sun Yat-sen University, 600 Tianhe Road, Guangzhou, Guangdong Province 510630, China.
Langenbecks Arch Surg. 2007 May;392(3):345-51. doi: 10.1007/s00423-006-0142-5. Epub 2007 Jan 19.
Cellular apoptosis plays an important role in ischemia-reperfusion (I/R) injury during organ transplantation. Synthetic small interference RNA (siRNA) targeting apoptotic receptor Fas has proven effective to protect mice against hepatitis and renal I/R injury. The objective of this study is to investigate the silencing impact of Fas siRNA to alleviate I/R injury in rat liver transplantation.
Rat hepatocytes (BRL cells) were transfected with three pairs of synthesized Fas siRNA; cells untreated and treated with GFP siRNA were taken as blank and siRNA control. The most effective Fas siRNA was chosen for in vivo experiments. Syngeneic orthotopic liver transplantation was performed in Fas siRNA group, siRNA control group, and blank control group of Sprague-Dawley rats. There were 25 pairs of rats in each group. siRNA transfection of donor rats was done with hydrodynamic injection method 48 h before liver procurement. Blood and liver samples were collected for evaluation of serum ALT levels, Fas protein and mRNA expression, and apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining, 1, 3, 6, 12, and 24 h after liver transplantation.
Fas siRNA2, which inhibited Fas gene expression much more than other siRNAs, was chosen for in vivo experiment. The serum ALT levels of Fas siRNA group were much less than those of blank and siRNA control groups 1, 3, 6, 12, and 24 h after blood reperfusion, indicating diminishing ischemia-reperfusion injury. Donor livers in Fas siRNA group had substantially less cell apoptosis. The expression of Fas mRNA and protein was reduced dramatically in the Fas siRNA group compared with the other two groups.
Fas-mediated apoptosis play an important role in I/R injury of rat liver transplantation. Silencing Fas by hydrodynamic injection of siRNA holds therapeutic promise to limit I/R injury.
细胞凋亡在器官移植的缺血再灌注（I/R）损伤中起重要作用。靶向凋亡受体Fas的合成小干扰RNA（siRNA）已被证明可有效保护小鼠免受肝炎和肾I/R损伤。本研究的目的是探讨Fas siRNA的沉默作用对减轻大鼠肝移植中I/R损伤的影响。
用三对合成的Fas siRNA转染大鼠肝细胞（BRL细胞）；未处理的细胞和用绿色荧光蛋白（GFP）siRNA处理的细胞分别作为空白对照和siRNA对照。选择最有效的Fas siRNA进行体内实验。在Fas siRNA组、siRNA对照组和空白对照组的Sprague-Dawley大鼠中进行同基因原位肝移植。每组有25对大鼠。在获取肝脏前48小时，采用水动力注射法对供体大鼠进行siRNA转染。肝移植后1、3、6、12和24小时，采集血液和肝脏样本，评估血清谷丙转氨酶（ALT）水平、Fas蛋白和mRNA表达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）染色检测细胞凋亡情况。
选择Fas siRNA2进行体内实验，其抑制Fas基因表达的能力比其他siRNA更强。血液再灌注后1、3、6、12和24小时，Fas siRNA组的血清ALT水平远低于空白对照组和siRNA对照组，表明缺血再灌注损伤减轻。Fas siRNA组供体肝脏的细胞凋亡明显减少。与其他两组相比，Fas siRNA组中Fas mRNA和蛋白的表达显著降低。
Fas介导的细胞凋亡在大鼠肝移植的I/R损伤中起重要作用。通过水动力注射siRNA沉默Fas对限制I/R损伤具有治疗前景。
Zotero 插件