药理学抑制巨噬细胞 Toll 样受体 4/核因子-κB 减轻横纹肌溶解症诱导的急性肾损伤。
Pharmacological Inhibition of Macrophage Toll-like Receptor 4/Nuclear Factor-kappa B Alleviates Rhabdomyolysis-induced Acute Kidney Injury.
机构信息
Kidney Research Institute, Department of Internal Medicine, Division of Nephrology, West China Hospital of Sichuan University, Chengdu, Sichuan 610041, China.
Division of Ultrasound, West China Hospital of Sichuan University, Chengdu, Sichuan 610041, China.
出版信息
Chin Med J (Engl). 2017 Sep 20;130(18):2163-2169. doi: 10.4103/0366-6999.213406.
BACKGROUND
: Acute kidney injury (AKI) is the most common and life-threatening systemic complication of rhabdomyolysis. Inflammation plays an important role in the development of rhabdomyolysis-induced AKI. This study aimed to investigate the kidney model of AKI caused by rhabdomyolysis to verify the role of macrophage Toll-like receptor 4/nuclear factor-kappa B (TLR4/NF-κB) signaling pathway.
METHODS
: C57BL/6 mice were injected with a 50% glycerin solution at bilateral back limbs to induce rhabdomyolysis, and CLI-095 or pyrrolidine dithiocarbamate (PDTC) was intraperitoneally injected at 0.5 h before molding. Serum creatinine levels, creatine kinase, the expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6, and hematoxylin and eosin stainings of kidney tissues were tested. The infiltration of macrophage, mRNA levels, and protein expression of TLR4 and NF-κB were investigated by immunofluorescence double-staining techniques, reverse transcriptase-quantitative polymerase chain reaction, and Western blotting, respectively. In vitro, macrophage RAW264.7 was stimulated by ferrous myoglobin; the cytokines, TLR4 and NF-κB expressions were also detected.
RESULTS
: In an in vivo study, using CLI-095 or PDTC to block TLR4/NF-κB, functional and histologic results showed that the inhibition of TLR4 or NF-κB alleviated glycerol-induced renal damages (P < 0.01). CLI-095 or PDTC administration suppressed proinflammatory cytokine (TNF-α, IL-6, and IL-1β) production and macrophage infiltration into the kidney (P < 0.01). Moreover, in an in vitro study, CLI-095 or PDTC suppressed myoglobin-induced expression of TLR4, NF-κB, and proinflammatory cytokine levels in macrophage RAW264.7 cells (P < 0.01).
CONCLUSION
: The pharmacological inhibition of TLR4/NF-κB exhibited protective effects on rhabdomyolysis-induced AKI by the regulation of proinflammatory cytokine production and macrophage infiltration.
背景
急性肾损伤（AKI）是横纹肌溶解最常见和最具威胁生命的全身并发症。炎症在横纹肌溶解诱导的 AKI 发展中起重要作用。本研究旨在探讨横纹肌溶解引起的 AKI 的肾脏模型，以验证巨噬细胞 Toll 样受体 4/核因子-κB（TLR4/NF-κB）信号通路的作用。
方法
向 C57BL/6 小鼠双侧后腿注射 50%甘油溶液诱导横纹肌溶解，在成型前 0.5 小时经腹腔注射 CLI-095 或吡咯烷二硫代氨基甲酸盐（PDTC）。检测血清肌酐水平、肌酸激酶、肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β和 IL-6 的表达以及肾组织的苏木精和伊红染色。通过免疫荧光双重染色技术、逆转录定量聚合酶链反应和 Western blot 分别检测巨噬细胞浸润、TLR4 和 NF-κB 的 mRNA 水平和蛋白表达。体外，用亚铁肌红蛋白刺激巨噬细胞 RAW264.7，检测细胞因子、TLR4 和 NF-κB 的表达。
结果
在体内研究中，使用 CLI-095 或 PDTC 阻断 TLR4/NF-κB，功能和组织学结果表明，TLR4 或 NF-κB 的抑制减轻了甘油诱导的肾损伤（P < 0.01）。CLI-095 或 PDTC 给药抑制促炎细胞因子（TNF-α、IL-6 和 IL-1β）的产生和巨噬细胞浸润到肾脏（P < 0.01）。此外，在体外研究中，CLI-095 或 PDTC 抑制肌红蛋白诱导的巨噬细胞 RAW264.7 细胞中 TLR4、NF-κB 和促炎细胞因子水平的表达（P < 0.01）。
结论
TLR4/NF-κB 的药理学抑制通过调节促炎细胞因子的产生和巨噬细胞浸润对横纹肌溶解诱导的 AKI 发挥保护作用。
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