Small Interfering RNA Targeting Toll-Like Receptor 9 Protects Mice against Polymicrobial Septic Acute Kidney Injury
Small Interfering RNA Targeting Toll-Like Receptor 9 Protects Mice against Polymicrobial Septic Acute Kidney Injury
复制标题
DOI：
10.1159/000346953
复制
发表时间：
2012-01-01
影响因子：
--
通讯作者：
Liu, Yueping
中科院分区：
文献类型：
--
作者：
Liu, Lixia;Li, Yong;Liu, Yueping
背景/目的：尽管最近的报告表明 Toll 样受体 (TLR) 9 与多种微生物脓毒症急性肾损伤 (AKI) 的发病机制相关，但目前尚不清楚肾脏 TLR9 是否以及如何参与多种微生物脓毒症 AKI 的发生。本研究旨在确定小鼠肾细胞中 TLR9 的表达是否与多种微生物脓毒症 AKI 的发生有关。方法：在培养的鼠巨噬细胞系（RAW264.7 细胞）中测试靶向 TLR9 的小干扰 RNA (siRNA) 的功效。在通过盲肠结扎和穿刺 (CLP) 诱导多种微生物脓毒症 AKI 之前，使用流体动力学方法将最有效的 siRNA 转染至小鼠体内。通过实时定量逆转录聚合酶链式反应 (RT-PCR) 和蛋白质印迹法在 RAW264.7 细胞和肾组织中确定 TLR9 敲低。在 CLP 后 6、12 和 24 小时时间点测定血清肌酐和血尿素氮 (BUN) 水平以及肾脏组织病理学评估，并在 24 小时研究肾细胞凋亡。还观察了小鼠4天和7天的存活率。结果：我们发现，尽管进行了液体和抗生素复苏，小鼠在我们的多种微生物败血症模型中仍出现 AKI，这与人类败血症类似。 TLR9 的 siRNA 成功沉默了 CLP 后肾 TLR9 基因和蛋白表达的诱导。有效沉默肾脏 TLR9 表达可减少肾细胞凋亡，减轻 AKI 的严重程度，并增加小鼠的存活率。结论：我们的数据表明 CLP 后小鼠肾组织中 TLR9 表达的诱导。我们的多种微生物脓毒症模型中的肾细胞凋亡和 AKI 依赖于 TLR9。因此，TLR9 可能在多种微生物脓毒症 AKI 的病理生理学中发挥关键作用。版权所有 (C) 2013 S. Karger AG，巴塞尔
Background/Aims: Although recent reports suggest that Toll-like receptor (TLR) 9 is associated with the pathogenesis of polymicrobial septic acute kidney injury (AKI), it is still unclear whether and how renal TLR9 is involved in the development of polymicrobial septic AKI. This study aimed to determine whether the expression of TLR9 in mouse renal cells is related to the development of polymicrobial septic AKI. Methods: The efficacy of small interfering RNA (siRNA) targeting TLR9 was tested in a cultured murine macrophage cell line (RAW264.7 cells). The most potent siRNA was transfected into mice using the hydrodynamic method prior to the induction of polymicrobial septic AKI being induced by cecal ligation and puncture (CLP). TLR9 knockdown was determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting in RAW264.7 cells and kidney tissues. The levels of serum creatinine and blood urea nitrogen (BUN) and the renal histopathology assessment were determined at 6-, 12-, and 24-hour time points after CLP, and renal cell apoptosis was studied at 24 h. The 4- and 7-day survival rates of mice were also observed. Results: We found that mice developed AKI in our model of polymicrobial sepsis, despite fluid and antibiotic resuscitation, which resembles human sepsis. siRNA to TLR9 successfully silenced the induction of renal TLR9 gene and protein expression following CLP. Effective silencing of renal TLR9 expression decreased renal cell apoptosis, mitigated the severity of AKI, and increased the survival of mice. Conclusions: Our data demonstrates the induction of TLR9 expression in mouse kidney tissue following CLP. Renal cell apoptosis and AKI in our model of polymicrobial sepsis are dependent on TLR9. Thus, TLR9 may play a critical role in the pathophysiology of polymicrobial septic AKI. Copyright (C) 2013 S. Karger AG, Basel