目的 探讨Toll样受体4（Toll like receptor 4，TLR4）/核转录因子κB（nuclear factor-κB，NF-κB）通路在二氧化硅（SiO2）所致大鼠矽肺模型肺泡巨噬细胞的脂质代谢中的作用。 方法 按随机数字表法将36只SD大鼠分为正常组、模型组、抑制剂组。模型组、抑制剂组采用一次性气管内缓慢滴注1 mL SiO2悬浮液进行造模。造模成功后抑制剂组大鼠每天定时耳缘静脉注射TAK-242（TLR4/NF-κB信号特异性抑制剂），剂量为0.5 mg/kg，正常组和模型组注射等剂量生理盐水。4周后处死大鼠收集支气管灌洗液（bronchial lavage fluid，BALF），ELISA测定各组大鼠BALF中白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的含量；提取BALF中的肺泡巨噬细胞进行培养，油红O染色观察各组大鼠肺泡巨噬细胞的脂滴形成；Western blot检测各组肺泡巨噬细胞中TLR4、MyD88、NF-κB的表达水平。 结果 与正常组相比，模型组、抑制剂组大鼠BALF中IL-6和TNF-α的含量，巨噬细胞中油红O阳性比例，巨噬细胞中TLR4、MyD88、NF-κB的表达升高，差异具有统计学意义（均P<0.05）；与模型组相比，抑制剂组大鼠上述指标降低，差异具有统计学意义（均P<0.05）。 结论 在SiO2所致大鼠矽肺模型中，TLR4/NF-κB信号参与肺泡巨噬细胞的脂质代谢的调控，抑制该信号的表达，能明显抑制矽肺的病理损伤。
Abstract
Objective To investigate the role of Toll-like receptor 4 (TLR4)/nuclear transcription factor κB (NF-κB) pathway in lipid metabolism of alveolar macrophages in the silicosis model of rats induced by silicon dioxide (SiO2). Methods According to the random number table method, 36 SD rats were randomly divided into a normal group, a model group and an inhibitor group. The model group and the inhibitor group were modeled by slowly instilling 1 mL of silica suspension into the disposable trachea. After successful modeling, the rats in the inhibitor group were injected with TAK-242 (TLR4/NF-κB signal specific inhibitor) daily with the dose of 0.5 mg/kg. The rats in the normal group and model group were injected an equal dose of normal saline. After 4 weeks, the rats were sacrificed to collect bronchial lavage fluid (BALF). Enzyme-linked immunosorbent assay (ELISA) was used to determine the content of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in BALF of each group of rats. Alveolar macrophages in BALF were extracted and cultured. Oil red O staining was used to observe the lipid droplet formation of macrophages in each group. Western blot was used to detect the expression levels of TLR4, MyD88, and NF-κB in macrophages. Results Compared with the normal group, the content of IL-6 and TNF-α in the BALF in the model group and the inhibitor group, the proportion of oil red O positive macrophages, the expression of TLR4, MyD88, and NF-κB in phagocytes all significantly increased, and the differences were statistically significant (all P<0.05). Compared with the model group, the content of IL-6 and TNF-α in BALF in the inhibitor group, the proportion of oil red O positive in macrophages, the expression of TLR4, MyD88, NF-κB in macrophages significantly reduced, the differences were statistically significant (all P<0.05). Conclusions In the silicosis model of rats caused by SiO2, TLR4/NF-κB signal is involved in the regulation of lipid metabolism of alveolar macrophages, and inhibit the expression of TLR4/NF-κB signal, which can significantly inhibit the pathological damage of silicosis.
关键词
Toll样受体4/核转录因子κB信号通路； /
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二氧化硅所致矽肺模型； /
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肺泡巨噬细胞； /
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脂质代谢
Key words
Toll like receptor4/ nuclear factor-κB signaling pathway; /
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Silicosis model induced by silicon dioxide (SiO2); /
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Alveolar macrophages; /
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Lipid metabolism
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基金
山西医科大学汾阳学院人才引进项目（2017D02）
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