环状RNA 0000157缺失通过微小RNA-149-5p/含溴结构域蛋白4通路保护人支气管上皮样细胞免受香烟烟雾提取物诱导的损伤。
Circular RNA 0000157 depletion protects human bronchial epithelioid cells from cigarette smoke extract-induced human bronchial epithelioid cell injury through the microRNA-149-5p/bromodomain containing 4 pathway.
作者信息
Song B, Wu S, Ye L, Jing Z, Cao J
机构信息
Department of Respiratory and Critical Care Medicine, The Second Hospital of Hebei Medical University, Shijiazhuang, China.
Department of Pharmacy, The Fourth Hospital of Shijiazhuang, Shijiazhuang, China.
出版信息
Hum Exp Toxicol. 2023 Jan-Dec;42:9603271231167581. doi: 10.1177/09603271231167581.
BACKGROUND
Circular RNA (circRNA) has been reported to regulate respiratory diseases. In the study, we aimed to elucidate the role of circ_0000157 in smoke-related chronic obstructive pulmonary disease (COPD) and the inner mechanism.
METHODS
COPD-like cell injury was induced by treating human bronchial epithelioid cells (16HBE) with cigarette smoke extract (CSE). The expression of circ_0000157, miR-149-5p, bromodomain containing 4 (BRD4), BCL2-associated x protein (Bax) and B-cell lymphoma-2 (Bcl-2) was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) or Western blotting. Enzyme-linked immunosorbent assay was performed to detect interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels. Malondialdehyde (MDA) production was detected by a lipid peroxidation MDA assay kit. Superoxide dismutase (SOD) activity was analyzed by a SOD activity assay kit.
RESULTS
Circ_0000157 and BRD4 expression were upregulated, while miR-149-5p expression was downregulated in the blood of smokers with COPD and CSE-induced 16HBE cells compared with control groups. CSE treatment inhibited 16HBE cell proliferation and induced cell apoptosis, inflammation, and oxidative stress; however, these effects were remitted when circ_0000157 expression was decreased. In addition, circ_0000157 acted as a miR-149-5p sponge and regulated CSE-caused 16HBE cell damage by targeting miR-149-5p. The overexpression of BRD4, a target gene of miR-149-5p, attenuated the inhibitory effects of miR-149-5p introduction on CSE-induced cell damage. Further, circ_0000157 modulated BRD4 expression by associating with miR-149-5p in CSE-treated 16HBE cells.
CONCLUSION
Circ_0000157 knockdown ameliorated CSE-caused 16HBE cell damage by targeting the miR-149-5p/BRD4 pathway, providing a potential therapeutic strategy for clinic intervention in COPD.
背景
据报道，环状RNA（circRNA）可调节呼吸系统疾病。在本研究中，我们旨在阐明circ_0000157在烟雾相关慢性阻塞性肺疾病（COPD）中的作用及其内在机制。
方法
用香烟烟雾提取物（CSE）处理人支气管上皮样细胞（16HBE），诱导出类似COPD的细胞损伤。通过定量实时聚合酶链反应（qRT-PCR）或蛋白质免疫印迹法分析circ_0000157、miR-149-5p、含溴结构域蛋白4（BRD4）、BCL2相关X蛋白（Bax）和B细胞淋巴瘤-2（Bcl-2）的表达。采用酶联免疫吸附测定法检测白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）水平。用脂质过氧化丙二醛检测试剂盒检测丙二醛（MDA）的生成量。用超氧化物歧化酶活性检测试剂盒分析超氧化物歧化酶（SOD）活性。
结果
与对照组相比，COPD吸烟者血液和CSE诱导的16HBE细胞中，circ_0000157和BRD4表达上调，而miR-149-5p表达下调。CSE处理抑制16HBE细胞增殖，诱导细胞凋亡、炎症和氧化应激；然而，当circ_0000157表达降低时，这些效应得到缓解。此外，circ_0000157作为miR-149-5p的海绵，通过靶向miR-149-5p调节CSE导致的16HBE细胞损伤。miR-149-5p的靶基因BRD4的过表达减弱了miR-149-5p导入对CSE诱导的细胞损伤的抑制作用。此外，在CSE处理的16HBE细胞中，circ_0000157通过与miR-149-5p结合来调节BRD4表达。
结论
敲低circ_0000157通过靶向miR-149-5p/BRD4通路改善CSE导致的16HBE细胞损伤，为COPD的临床干预提供了一种潜在的治疗策略。
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