RNA interference targeting hypoxia inducible factor 1alpha reduces post-operative adhesions in rats.
RNA interference targeting hypoxia inducible factor 1alpha reduces post-operative adhesions in rats.
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DOI：
10.1016/j.jss.2006.07.045
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发表时间：
2007-08
期刊：
影响因子：
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通讯作者：
T. Segura;H. Schmokel;J. Hubbell
中科院分区：
文献类型：
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作者：
T. Segura;H. Schmokel;J. Hubbell
目的探讨RNA干扰（siRNAs）下调缺氧诱导因子1α（HIF-1α）和纤溶酶原激活物抑制剂1（派-1）基因表达以预防腹腔粘连的发生。对于体内实验，对57只白色雌性大鼠进行手术以使子宫角产生缺血性和血清性损伤，并用盐水、siRNAamin A/C（阴性对照）、siRNAHIF-1α、siRNAAPAI-1或siRNAHIF-1α加siRNAAPAI-1处理。使用阳离子聚合物聚（乙烯亚胺）（PEI）作为体内递送所有siRNA的递送载体。术后8天由不知情的外科医生分析粘连。在用siRNA体外转染后，对于所有测试的siRNA，获得至少69%的基因下调。在体外，siRNA下调HIF-1α和派-1表达或同时下调HIF-1α和PAI-1表达均能显著降低派-1蛋白活性（P < 0.05）。在子宫角损伤后给予4 nmol siRNAHIF-1α/PEI复合物可使术后粘连形成减少52%（P < 0.05）。递送4 nmol siRNAPAI-1/PEI复合物和同时递送2 nmol siRNAHIF-1α加2 nmol siRNAPAI-1分别导致腹部粘连减少36%和42%，当与盐水对照直接比较时，结论腹腔注射siRNA/PEI复合物可预防术后盆腹腔粘连。
BACKGROUNDTo investigate the use of RNA interference mediated gene down-regulation targeting hypoxia inducible factor 1α (HIF-1α) and plasminogen activator inhibitor 1 (PAI-1) in an effort to prevent abdominal adhesion formation.MATERIALS AND METHODSReal time PCR and a PAI-1 protein activity assay were used in vitro to determine the efficacy of small interfering RNAs (siRNAs). For in vivo experiments, 57 white female rats were operated to generate ischemic and serosal injury to the uterine horns, and treated with saline, siRNALamin A/C(negative control), siRNAHIF-1α, siRNAPAI-1, or siRNAHIF-1αplus siRNAPAI-1. The cationic polyer poly(ethylenimine) (PEI) was used as the delivery vehicle for all siRNAs delivered in vivo. Adhesions were analyzed by a blinded surgeon 8 days post-surgery.RESULTSAfter in vitro transfection with siRNA, at least 69% gene down-regulation was obtained for all siRNAs tested. In vitro siRNA-mediated down-regulation of HIF-1α, PAI-1 or their simultaneous delivery resulted in a significant decrease of PAI-1 protein activity (at least P < 0.05). Administration of 4 nmol siRNAHIF-1α/PEI complexes after injury to the uterine horns achieved a statistical reduction of post-operative adhesion formation with a reduction by 52% (P < 0.05). Delivery of 4 nmol siRNAPAI-1/PEI complexes and the simultaneous delivery of 2 nmol siRNAHIF-1αplus 2 nmol siRNAPAI-1, resulted in a reduction of abdominal adhesion by 36% and 42%, respectively, with the reduction being statistically significant when compared directly to the saline control (P < 0.01).CONCLUSIONThese data show that administration of siRNA/PEI complexes within the peritoneal cavity can be used to prevent post-operative abdominopelvic adhesions.