Department of Occupational and Environmental Health, School of Public Health, Jilin University, Changchun, China.
Preventive and health care, Xianlin Health Service Center of Yuhang District in Hangzhou City, Hangzhou, China.
Environ Toxicol. 2023 Sep;38(9):2256-2270. doi: 10.1002/tox.23865. Epub 2023 Jun 19.
PM can cause airway inflammation and promote the excessive secretion of mucin 5ac (Muc5ac), which can further induce many respiratory diseases. Antisense non-coding RNA in the INK4 locus (ANRIL) might regulate the inflammatory responses mediated by nuclear factor kappa-B (NF-κB) signaling pathway. Beas-2B cells were used to clarify the role of ANRIL in the secretion of Muc5ac induced by PM . The siRNA was used to silence ANRIL expression. Normal and gene silenced Beas-2B cells were respectively exposed to different doses of PM for 6, 12, and 24 h. The survival rate of Beas-2B cells was detected by methyl thiazolyl tetrazolium (MTT) assay. Tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and Muc5ac levels were determined by enzyme linked immunosorbent assay (ELISA). The expression levels of NF-κB family genes and ANRIL were detected by real time polymerase chain reaction (PCR). The levels of NF-κB family proteins and NF-κB family phosphorylated proteins were determined using Western blot. Immunofluorescence experiments were performed to observe the nuclear transposition of RelA. PM exposure increased the levels of Muc5ac, IL-1β and TNF-α, and ANRIL gene expression (p < .05). With the dose and time of PM exposure increasing the protein levels of inhibitory subunit of nuclear factor kappa-B alpha (IκB-α), RelA, and NF-κB1 decreased, the protein levels of phosphorylated RelA (p-RelA) and phosphorylated NF-κB1 (p-NF-κB1) increased, and RelA nuclear translocation increased, which indicated that the NF-κB signaling pathway was activated (p < .05). Silencing ANRIL could decrease the levels of Muc5ac, IL-1β, TNF-α, decrease NF-κB family genes expression, inhibit the degradation of IκB-α and the activation of NF-κB pathway (p < .05). ANRIL played a regulatory role in the secretion of Muc5ac and the inflammation induced by atmospheric PM via NF-κB pathway in Beas-2B cells. ANRIL could be a target for prevention and treatment of the respiratory diseases caused by PM .
PM 可引起气道炎症，并促进黏蛋白 5ac（Muc5ac）的过度分泌，这可能进一步导致多种呼吸道疾病。INK4 基因座中的反义非编码 RNA（ANRIL）可能通过核因子-κB（NF-κB）信号通路调节炎症反应。Beas-2B 细胞被用来阐明 ANRIL 在 PM 诱导的 Muc5ac 分泌中的作用。用 siRNA 沉默 ANRIL 表达。分别用不同剂量的 PM 处理正常和基因沉默的 Beas-2B 细胞 6、12 和 24 小时。噻唑蓝（MTT）比色法检测 Beas-2B 细胞的存活率。酶联免疫吸附试验（ELISA）测定肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和 Muc5ac 水平。实时聚合酶链反应（PCR）检测 NF-κB 家族基因和 ANRIL 的表达水平。采用 Western blot 检测 NF-κB 家族蛋白和 NF-κB 家族磷酸化蛋白的水平。免疫荧光实验观察 RelA 的核易位。PM 暴露增加了 Muc5ac、IL-1β 和 TNF-α 的水平和 ANRIL 基因表达（p <.05）。随着 PM 暴露剂量和时间的增加，核因子 kappa-B 抑制亚单位 alpha（IκB-α）、RelA 和 NF-κB1 的蛋白水平降低，磷酸化 RelA（p-RelA）和磷酸化 NF-κB1（p-NF-κB1）的蛋白水平升高，RelA 核易位增加，表明 NF-κB 信号通路被激活（p <.05）。沉默 ANRIL 可降低 Muc5ac、IL-1β、TNF-α 水平，降低 NF-κB 家族基因表达，抑制 IκB-α 降解和 NF-κB 通路激活（p <.05）。ANRIL 通过 NF-κB 通路在 Beas-2B 细胞中对大气 PM 诱导的 Muc5ac 分泌和炎症起调节作用。ANRIL 可能是预防和治疗 PM 引起的呼吸道疾病的靶点。
