Biochanin A protects against PM2.5-induced acute pulmonary cell injury by interacting with the target protein MEK5.
Biochanin A protects against PM2.5-induced acute pulmonary cell injury by interacting with the target protein MEK5.
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DOI：
10.1039/c9fo01382b
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发表时间：
2019-11
期刊：
影响因子：
6.1
通讯作者：
Zhaohui Xue;Junyu Wang;Wancong Yu;Dan Li;Yixia Zhang;Fang Wan;X. Kou
中科院分区：
文献类型：
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作者：
Zhaohui Xue;Junyu Wang;Wancong Yu;Dan Li;Yixia Zhang;Fang Wan;X. Kou
流行病学研究表明，暴露于环境细颗粒物（PM2.5）与心肺疾病风险增加有关。MEK5/ERK5和NF-κB信号通路与急性肺细胞损伤（APCI）的调控密切相关，可能在其潜在的病理生理机制中发挥重要作用。相关研究表明，生物茶素A （Biochanin A， BCA）可有效干扰APCI，但其发生的潜在机制尚不完全清楚。此前，基于蛋白质组学和生物信息学研究，我们发现MEK5在介导BCA对pm2.5肺毒性的缓解作用中发挥了不可或缺的作用。因此，我们利用A549腺癌人肺泡基底上皮细胞（A549细胞），结合western blot和qRT-PCR对BCA诱导的保护性信号通路进行了研究，发现MEK5/ERK5和NF-κB均参与了APCI对PM2.5的响应，且MEK5/ERK5正激活了NF-κB及其下游细胞调节因子。BCA显著抑制pm2.5诱导的MEK5/ERK5表达上调以及NF-κB的磷酸化和活化。此外，由于MEK5/ERK5蛋白结构的特异性，利用分子对接相关技术分析了BCA与MEK5的结合位点和结合模式，结果表明BCA与MEK5的PB1结构域及其激酶结构域之间存在稳定的氢键。BCA与MEK5形成稳定的复合物，对MEKK2/3-MEK5-ERK5三元相互作用、p62/α pkc介导的NF-κB调控、抑制MEK5靶蛋白磷酸化具有潜在作用。因此，我们的研究表明，MEK5是APCI对PM2.5暴露的细胞内信号传导的重要调节因子。BCA可能通过靶向MEK5抑制MEK5/ERK5/NF-κB信号通路的激活而发挥抗apci活性。
Epidemiological studies have shown that exposure to ambient fine particulate matter (PM2.5) is associated with an increased risk for cardiopulmonary diseases. The MEK5/ERK5 and NF-κB signaling pathways are closely related to the regulation of acute pulmonary cell injury (APCI) and may play an important role in the underlying pathophysiological mechanisms. Related studies have shown that Biochanin A (BCA) effectively interferes with APCI, but the underlying mechanism through which this occurs is not fully understood. Previously, based on proteomic and bioinformatic research, we found the indispensable role of MEK5 in mediating remission effects of BCA against PM2.5-induced lung toxicity. Therefore, using A549 adenocarcinoma human alveolar basal epithelial cells (A549 cells), we combined western blot and qRT-PCR to study the protective signaling pathways induced by BCA, indicating that MEK5/ERK5 and NF-κB are both involved in mediating APCI in response to PM2.5, and MEK5/ERK5 positively activated NF-κB and its downstream cellular regulatory factors. BCA significantly suppressed PM2.5-induced upregulation of MEK5/ERK5 expression and phosphorylation and activation of NF-κB. Furthermore, due to the specificity of the MEK5/ERK5 protein structure, the binding sites and binding patterns of BCA and MEK5 were analyzed using molecular docking correlation techniques, which showed that there are stable hydrogen bonds between BCA and the PB1 domain of MEK5 as well as its kinase domain. BCA forms a stable complex with MEK5, which has potential effects on MEKK2/3-MEK5-ERK5 ternary interactions, p62/αPKC-mediated NF-κB regulation, and inhibition of MEK5 target protein phosphorylation. Therefore, our study suggests that MEK5 is an important regulator of intracellular signaling of APCI in response to PM2.5 exposure. BCA may exert anti-APCI activity by targeting MEK5 to inhibit activation of the MEK5/ERK5/NF-κB signaling pathway.