TLR4／NF-κB信号通路在SiO2所致矽肺模型肺泡巨噬细胞脂质代谢调控中的作用#br#

doi:10.13418/j.issn.1001-165x.2022.3.12

摘要/Abstract

摘要： 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。

关键词: Toll样受体4/核转录因子κB信号通路； , , 二氧化硅所致矽肺模型； , , 肺泡巨噬细胞； , , 脂质代谢

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.

Key words: Toll like receptor4/ nuclear factor-κB signaling pathway; , , Silicosis model induced by silicon dioxide (SiO2); , , Alveolar macrophages; , , Lipid metabolism

中图分类号:

R563.9

赵婧, 焦卓亚, 王娟, 刘璐. TLR4／NF-κB信号通路在SiO2所致矽肺模型肺泡巨噬细胞脂质代谢调控中的作用#br#[J]. 中国临床解剖学杂志, 2022, 40(3): 309-314.

Zhao Jing, Jiao Zhuoya, Wang Juan, Liu Lu. The role of TLR4/NF-κB signaling pathway in the regulation of lipid metabolism of alveolar macrophages in a model of silicosis induced by SiO2[J]. Chinese Journal of Clinical Anatomy, 2022, 40(3): 309-314.

参考文献 17

[1]	Bukovitz B, Meiman J, Anderson H, et al. Silicosis: diagnosis and medicolegal implications[J]. J Forensic Sci, 2019, 64(5): 1389-1398. DOI: 10.1111/1556-4029.14048.
[2]	Ophir N, Shai AB, Korenstein R, et al. Functional, inflammatory and interstitial impairment due to artificial stone dust ultrafine particles exposure[J]. Occup Environ Med, 2019, 76(12): 875-879. DOI: 10.1136/oemed-2019-105711.
[3]	Wang D, Hao CF, Zhang L, et al. Exosomal miR-125a-5p derived from silica-exposed macrophages induces fibroblast transdifferentiation[J]. Ecotoxicol Environ Saf, 2020, 192(1): 110253-110263. DOI: 10.1016/j.ecoenv.2020.110253.
[4]	Zhang ZQ, Zhang CZ, Shao B, et al. Effects of abnormal expression of fusion and fission genes on the morphology and function of lung macrophage mitochondria in SiO2-induced silicosis fibrosis in rats in vivo[J]. Toxicol Lett, 2019, 312(9): 181-187. DOI: 10.1016/j.toxlet.2019.04.029.
[5]	Hou XM, Summer R, Chen ZY, et al. Lipid uptake by alveolar macrophages drives fibrotic responses to silica dust[J]. Sci Rep, 2019, 9(1): 399-412. DOI: 10.1038/s41598-018-36875-2.
[6]	Hegde B, Bodduluri SR, Satpathy SR, et al. Inflammasome-independent leukotriene B4 production drives crystalline silica-induced sterile inflammation[J]. J Immunol, 2018, 200(10): 3556-3567. DOI: 10.4049/jimmunol.1701504.
[7]	Pan W, Xu XH, Wang Y, et al. Interleukin-35 reduces inflammation in acute lung injury through inhibiting TLR4/NF-κB signaling pathways[J]. Exp Ther Med, 2020, 19(3): 1695-1700. DOI: 10.3892/etm. 2020. 8407.
[8]	陆青兰, 黄世稳, 韦雪平, 等. 黄根多糖对大鼠矽肺纤维化的作用[J]. 中草药, 2020, 51(4): 1031-1036. DOI: 10.7501/j.issn.0253-2670. 2020. 04.029.
[9]	颜君, 于春锐, 孙立新, 等. 盐酸戊乙奎醚对大鼠呼吸机相关性肺损伤时气道MUC5AC表达的影响及其与TLR4/MyD88信号通路的关系[J]. 中华麻醉学杂志, 2019, 39(10): 1248-1252. DOI: 10.3760∕cma.j.issn.0254-1416.2019.10.025.
[10]	Giannakis N, Sansbury BE, Patsalos A, et al. Dynamic changes to lipid mediators support transitions among macrophage subtypes during muscle regeneration[J]. Nat Immunol, 2019, 20(5): 626-636. DOI: 10.1038/s41590-019-0356-7.
[11]	黄鸿波, 庄锡彬, 郑锦阳, 等. 矽肺合并机化性肺炎119例临床诊断与治疗[J]. 环境与职业医学, 2019, 36(8): 761-766. DOI: 10.13213/j.cnki.jeom.2019.19125.
[12]	赵娜, 吴洁, 吴奇峰, 等. 矽肺患者血清甘露聚糖结合凝集素与Th17/Treg关系[J]. 中国职业医学, 2019, 46(3): 263-268. DOI: 10.11763/j.issn.2095-2619.2019.03.001.
[13]	Du ST, Li C, Lu YP, et al. Dioscin alleviates crystalline silica-induced pulmonary inflammation and fibrosis through promoting alveolar macrophage autophagy[J]. Theranostics, 2019, 9(7): 1878-1892. DOI: 10.7150/thno.29682.
[14]	Fang SC, Guo HF, Cheng YS, et al. circHECTD1 promotes the silica-induced pulmonary endothelial-mesenchymal transition via HECTD1[J]. Cell Death Dis, 2018, 9(3): 396-412. DOI: 10.1038/s41419-018-0432-1.
[15]	Jiang KF, Yang J, Guo S, et al. Peripheral circulating exosome-mediated delivery of miR-155 as a novel mechanism for acute lung inflammation[J]. Mol Ther, 2019, 27(10): 1758-1771. DOI: 10.1016/j.ymthe. 2019. 07.003.
[16]	Liu J, Chen T, Lei P, et al. Exosomes released by bone marrow mesenchymal stem cells attenuate lung injury induced by intestinal ischemia reperfusion via the TLR4/NF-κB pathway[J]. Int J Med Sci, 2019, 16(9): 1238-1244. DOI: 10.7150/ijms.35369.
[17]	Wang YF, Zhang XR, Tian JM, et al. Sevoflurane alleviates LPS‑induced acute lung injury via the microRNA‑27a‑3p/TLR4/MyD88/NF‑κB signaling pathway[J]. Int J Mol Med, 2019, 44(2): 479-490. DOI: 10.3892/ijmm.2019.4217.