Chronic kidney disease (CKD) is associated with endothelial dysfunctions thus prompting links between microcirculation (MC), inflammation and major cardiovascular risk factors. We have previously reported that siRNA-silencing of CD40 (siCD40) reduced atherosclerosis (ATH) progression. Here, we have deepened on the effects of the siCD40 treatment by evaluating retrospectively, in stored kidneys from the siCD40 treated ApoE−/− mice, the renal microcirculation (measured as the density of peritubular capillaries), macrophage infiltration and NF-κB activation. Kidneys were isolated after 16 weeks of treatment with the anti-CD40 siRNA (siCD40), with a scrambled control siRNA (siSC) or with PBS (Veh. group). Renal endothelium, infiltrating macrophages and activated NF-κB in endothelium were identified by immunohistochemistry, while the density of stained peritubular capillaries was quantified by image analysis. ATH was associated with a reduction in renal MC, an effect reversed by the anti-CD40 siRNA treatment (3.8 ± 2.7% in siCD40; vs. 1.8 ± 0.1% in siSC; or 1.9 ± 1.6% in Veh.; p < 0.0001). Furthermore, siCD40 treatment reduced the number of infiltrating macrophages compared to the SC group (14.1 ± 5.9 cells/field in siCD40; vs. 37.1 ± 17.8 cells/field in siSC; and 1.3 ± 1.7 cells/field in Veh.; p = 0.001). NF-κB activation also peaked in the siSC group, showing lower levels in the siCD40 and Veh. groups (63 ± 60 positive cells/section in siCD40; vs. 152 ± 44 positive cells/section in siSC; or 26 ± 29 positive cells/section in veh.; p = 0.014). Lastly, serum creatinine was also increased in the siCD40 (3.4 ± 3.3 mg/dL) and siSC (4.6 ± 3.0 mg/dL) groups when compared with Veh. (1.1 ± 0.9 mg/dL, p = 0.1). Anti-CD40 siRNA therapy significantly increased the density of peritubular capillaries and decreased renal inflammation in the ATH model. These data provide a physiological basis for the development of renal diseases in patients with ATH. Furthermore, our results also highligth renal off-target effects of the siRNA treatment which are discussed.

中文翻译：

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抗 CD40 siRNA 疗法的双刃剑：它增加了肾脏微毛细血管的密度，但有利于在 ApoE-/- 小鼠的肾脏中产生炎症环境

慢性肾病 (CKD) 与内皮功能障碍有关，因此促进了微循环 (MC)、炎症和主要心血管危险因素之间的联系。我们之前曾报道过 CD40 (siCD40) 的 siRNA 沉默可减少动脉粥样硬化 (ATH) 的进展。在这里，我们通过回顾性评估，在 siCD40 处理的 ApoE-/- 小鼠的储存肾脏中，加深了 siCD40 治疗的效果，肾微循环（测量为管周毛细血管的密度）、巨噬细胞浸润和 NF-κB 活化。在用抗 CD40 siRNA (siCD40)、乱序对照 siRNA (siSC) 或 PBS (Veh. group) 治疗 16 周后分离肾脏。通过免疫组织化学鉴定肾内皮、浸润性巨噬细胞和内皮中活化的 NF-κB，而染色的管周毛细血管的密度通过图像分析进行量化。ATH 与肾 MC 的降低有关，抗 CD40 siRNA 治疗逆转了这一效应（siCD40 为 3.8 ± 2.7%；siSC 为 1.8 ± 0.1%；Veh. 为 1.9 ± 1.6%；p < 0.0001） . 此外，与 SC 组相比，siCD40 治疗减少了浸润巨噬细胞的数量（siCD40 中 14.1 ± 5.9 个细胞/视野；与 siSC 中 37.1 ± 17.8 个细胞/视野相比；Veh 中 1.3 ± 1.7 个细胞/视野；p = 0.001 ）。NF-κB 激活也在 siSC 组中达到峰值，显示 siCD40 和 Veh 的水平较低。组（siCD40 中 63 ± 60 个阳性细胞/切片；与 siSC 中 152 ± 44 个阳性细胞/切片相比；或车辆中 26 ± 29 个阳性细胞/切片；p = 0.014）。最后，血清肌酐在 siCD40 (3.4 ± 3.3 mg/dL) 和 siSC (4.6 ± 3. 0 mg/dL) 组与 Veh 相比。（1.1 ± 0.9 毫克/分升，p = 0.1）。在 ATH 模型中，抗 CD40 siRNA 治疗显着增加了管周毛细血管的密度并减少了肾脏炎症。这些数据为 ATH 患者肾脏疾病的发展提供了生理学基础。此外，我们的结果还强调了讨论的 siRNA 治疗的肾脱靶效应。