Xu Jianguo, Li Xin, Zeng Xiaoli, Bao Hairong, Liu Xiaoju
Department of Gerontal Respiratory Medicine, The First Hospital of Lanzhou University, Lanzhou, 730000, China.
Curr Med Chem. 2024 Oct 3. doi: 10.2174/0109298673346794241001110826.
Tobacco smoke is an important inducer of airway epithelial cell aging. Punicalagin(PCG) is a natural anti-aging compound. The effect of PCG on tobacco smoke-induced airway epithelial cell senescence is unknown.
Our study investigated whether PCG can treat the human bronchial epithelial cell line (BEAS-2B) aging by inhibiting the protease-activated receptor 2 (PAR2)/m- TOR pathway.
Bioinformatics techniques were used to analyze the potential biological functions of PAR2. Molecular dynamics evaluated the binding ability of PCG and PAR2. The CCK8 assay was used to detect the cytotoxicity of CSE and PCG. The activity of the PAR2/mTOR pathway and the expression of the characteristic aging markers p16, p21, and SIRT1 are detected by qRT-PCR and Western blotting. Cell senescence was observed by Senescence-associated β-galactosidase (SA-β-gal) staining. The senescence-associated secretory phenotype (SASP): concentrations of interleukin IL-6, IL-8, and TNF- α were detected by ELISA.
The GSE57148 bioinformatics analysis dataset showed that PAR2 regulates lung senescence through the mTOR signaling pathway. Molecular dynamics results found that PCG and PAR2 had a strong and stable binding force. CSE induces BEAS-2B cell senescence and activates the PAR2/mTOR pathway. Inhibition of PAR2 mitigated the senescence changes. In addition, PCG's pretreatment can significantly alleviate CSE-induced BEAS-2B cell senescence while inhibiting the PAR2/mTOR pathway.
PCG has a therapeutic effect on the senescence of airway epithelial cells.
烟草烟雾是气道上皮细胞衰老的重要诱导因素。没食子酸丙酯（PCG）是一种天然的抗衰老化合物。PCG对烟草烟雾诱导的气道上皮细胞衰老的影响尚不清楚。
本研究探讨PCG是否能通过抑制蛋白酶激活受体2（PAR2）/m-TOR通路来治疗人支气管上皮细胞系（BEAS-2B）衰老。
采用生物信息学技术分析PAR2的潜在生物学功能。分子动力学评估PCG与PAR2的结合能力。采用CCK8法检测香烟烟雾提取物（CSE）和PCG的细胞毒性。通过qRT-PCR和蛋白质免疫印迹法检测PAR2/mTOR通路的活性以及衰老特征标志物p16、p21和SIRT1的表达。通过衰老相关β-半乳糖苷酶（SA-β-gal）染色观察细胞衰老情况。采用酶联免疫吸附测定（ELISA）法检测衰老相关分泌表型（SASP）：白细胞介素IL-6、IL-8和肿瘤坏死因子-α的浓度。
GSE57148生物信息学分析数据集显示，PAR2通过mTOR信号通路调节肺衰老。分子动力学结果发现，PCG与PAR2具有强大而稳定的结合力。CSE诱导BEAS-2B细胞衰老并激活PAR2/mTOR通路。抑制PAR2可减轻衰老变化。此外，PCG预处理可显著减轻CSE诱导的BEAS-2B细胞衰老，同时抑制PAR2/mTOR通路。
PCG对气道上皮细胞衰老具有治疗作用。
