Yang Cheng, Zhao Tian, Zhao Zitong, Jia Yichen, Li Long, Zhang Yufang, Song Mangen, Rong Ruiming, Xu Ming, Nicholson Michael L, Zhu Tongyu, Yang Bin
Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
Department of Nephrology, Affiliated Hospital of Nantong University, Medical Research Centre, Medical School, University of Nantong, Nantong, China.
Mol Ther. 2014 Oct;22(10):1817-28. doi: 10.1038/mt.2014.111. Epub 2014 Jun 16.
The naked small interfering RNA (siRNA) of caspase-3, a key player in ischemia reperfusion injury, was effective in cold preserved and hemoreperfused kidneys, but not autotransplanted kidneys in our porcine models. Here, chemically modified serum stabilized caspase-3 siRNAs were further evaluated. The left kidney was retrieved and infused by University of Wisconsin solution with/without 0.3 mg caspase-3 or negative siRNA into the renal artery for 24-hour cold storage (CS). After an intravenous injection of 0.9 mg siRNA and right-uninephrectomy, the left kidney was autotransplanted for 2 weeks. The effectiveness of caspase-3 siRNA was confirmed by caspase-3 knockdown in the post-CS and/or post-transplant kidneys with reduced apoptosis and inflammation, while the functional caspase-3 siRNA in vivo was proved by detected caspase-3 mRNA degradation intermediates. HMGB1 protein was also decreased in the post-transplanted kidneys; correlated positively with renal IL-1β mRNA, but negatively with serum IL-10 or IL-4. The minimal off-target effects of caspase-3 siRNA were seen with favorable systemic responses. More importantly, renal function, associated with active caspase-3, HMGB1, apoptosis, inflammation, and tubulointerstitial damage, was improved by caspase-3 siRNA. Taken together, the 2-week autotransplanted kidneys were protected when caspase-3 siRNA administrated locally and systemically, which provides important evidence for future clinical trials.
半胱天冬酶-3的裸小干扰RNA（siRNA）是缺血再灌注损伤中的关键因子，在我们的猪模型中，它对冷保存和血液灌注的肾脏有效，但对自体移植肾脏无效。在此，对化学修饰的血清稳定化半胱天冬酶-3 siRNAs进行了进一步评估。取出左肾，用含或不含0.3 mg半胱天冬酶-3或阴性siRNA的威斯康星大学溶液经肾动脉灌注，进行24小时冷保存（CS）。静脉注射0.9 mg siRNA并切除右侧单肾后，将左肾自体移植2周。通过在冷保存后和/或移植后的肾脏中敲低半胱天冬酶-3，减少细胞凋亡和炎症，证实了半胱天冬酶-3 siRNA的有效性，同时通过检测半胱天冬酶-3 mRNA降解中间体证明了其在体内的功能性。移植后肾脏中的高迁移率族蛋白B1（HMGB1）蛋白也减少；与肾脏白细胞介素-1β mRNA呈正相关，但与血清白细胞介素-10或白细胞介素-4呈负相关。半胱天冬酶-3 siRNA的脱靶效应最小，全身反应良好。更重要的是，半胱天冬酶-3 siRNA改善了与活性半胱天冬酶-3、HMGB1、细胞凋亡、炎症和肾小管间质损伤相关的肾功能。综上所述，当局部和全身给予半胱天冬酶-3 siRNA时，2周的自体移植肾脏得到了保护，这为未来的临床试验提供了重要证据。
