基于TMT的定量蛋白质组学揭示淫羊藿苷II通过抑制OVA诱导的慢性哮喘小鼠中LAMP2、CTSD和CTSS的表达从而对气
2023/09/21
基于TMT的定量蛋白质组学揭示淫羊藿苷II通过抑制OVA诱导的慢性哮喘小鼠中LAMP2、CTSD和CTSS的表达从而对气道炎症和重塑起到保护作用
摘要背景：哮喘是一种气道慢性炎症性疾病，具有气流受限、气道炎症和重塑的典型病理特征。淫羊藿苷II（IS）来源于草药淫羊藿，具有抗炎作用。然而，IS在哮喘中特异性靶向分子表达的潜在机制尚未完全了解，IS是否可以抑制重塑和上皮-间质转化（EMT）仍不清楚。
目的：本研究旨在阐明IS对减轻哮喘气道炎症和重塑的治疗效果，并通过基于TMT的定量蛋白质组学说明IS调节的特定途径和靶蛋白。
研究设计和方法：用卵清蛋白（OVA）致敏构建慢性哮喘小鼠模型，然后激发8周。检测肺功能、支气管肺泡灌洗液（BALF）中的白细胞计数、肺组织病理学、炎症和纤维化细胞因子以及EMT的标志物。选择肺组织进行基于TMT的定量蛋白质组学分析，以探索IS调节的蛋白质。
结果：IS有助于缓解气道高反应性（AHR），表现为RL下降、Cdyn增加。本研究观察到经IS治疗后BALF中白细胞计数、炎症细胞因子水平和支气管周围炎症浸润明显下调。杯状细胞增生、粘液分泌和支气管周围胶原沉积减弱，BALF中TGF-β和MMP-9水平下降。此外，IS诱导肺组织中Occludin和E-cadherin的升高以及N-cadherin和α-SMA的下降。上述结果证明了IS对气道炎症、气道重塑和EMT的保护作用。为了进一步研究IS在哮喘治疗中的潜在机制，本研究进行了基于TMT的定量蛋白质组学，并鉴定了由IS调节的102个重叠的DEP。KEGG富集分析表明这些DEP富含溶酶体、吞噬体和自噬，其中LAMP2、CTSD和CTSS是常见的DEP。WB、q-PCR和IHC结果证明了这些蛋白质的表达改变。此外，IS可以随着p62表达的增加而抑制Beclin-1和LC3B的表达，从而抑制自噬。
结论：本研究表明IS可以改善OVA诱导的慢性哮喘小鼠的AHR、气道炎症、气道重塑和EMT。本研究首次揭示了抑制自噬中LAMP2、CTSD和CTSS的表达有助于IS对哮喘的治疗效果。
(中日友好医院呼吸与危重症医学科 张婧媛 摘译 林江涛 审校)
(Phytomedicine. 2023 Sep;118:154941. doi: 10.1016/j.phymed.2023.154941.)
(Phytomedicine. 2023 Sep;118:154941. doi: 10.1016/j.phymed.2023.154941.)
TMT-based quantitative proteomics revealed protective efficacy of Icariside II against airway inflammation and remodeling via inhibiting LAMP2, CTSD and CTSS expression in OVA-induced chronic asthma mice
Yaolong Zhou, Xi Huang, Hang Yu, Hanlin Shi, Mengmeng Chen, Jingrong Song, Weifeng Tang, Fangzhou Teng, Congcong Li, La Yi, Xueyi Zhu, Na Wang, Ying Wei, Tulake Wuniqiemu, Jingcheng Dong
Abstract
Background:Asthma is a chronic inflammatory disorder in airways with typical pathologic features of airflow limitation, airway inflammation and remodeling. Icariside II (IS), derived from herbal medicine Herba Epimedii, exerts an anti-inflammatory property. However, underlying mechanisms with specifically targeted molecular expression by IS in asthma have not been fully understood, and whether IS could inhibit remodeling and EMT still remains unclear.
Purpose:The study aimed to clarify therapeutic efficacy of IS for attenuating airway inflammation and remodeling in asthma, and illustrate IS-regulated specific pathway and target proteins through TMT-based quantitative proteomics.
Study design and methods:Murine model of chronic asthma was constructed with ovalbumin (OVA) sensitization and then challenge for 8 weeks. Pulmonary function, leukocyte count in bronchoalveolar lavage fluid (BALF), lung histopathology, inflammatory and fibrotic cytokines, and markers of epithelial-mesenchymal transition (EMT) were evaluated. TMT-based quantitative proteomics were performed on lung tissues to explore IS-regulated proteins.
RESULTS:IS contributed to alleviative airway hyperresponsiveness (AHR) evidenced by declined RL and increased Cdyn. After IS treatment, we observed a remarked down-regulation of leukocyte count, inflammatory cytokines in BALF, and peribronchial inflammation infiltration. Goblet cell hyperplasia, mucus secretion and peribronchial collagen deposition were attenuated, with the level of TGF-β and MMP-9 in BALF declined. Furthermore, IS induced a rise of Occludin and E-cadherin and a decline of N-cadherin and α-SMA in lung tissues. These results proved the protective property of IS against airway inflammation, remodeling and EMT. To further investigate underlying mechanisms of IS in asthma treatment, TMT-based quantitative proteomics were performed and 102 overlapped DEPs regulated by IS were identified. KEGG enrichment exhibited these DEPs were enriched in lysosome, phagosome and autophagy, in which LAMP2, CTSD and CTSS were common DEPs. WB, q-PCR and IHC results proofed expressional alteration of these proteins. Besides, IS could decrease Beclin-1 and LC3B expression with increasing p62 expression thus inhibiting autophagy.
CONCLUSION:The study demonstrated IS could ameliorate AHR, airway inflammation, remodeling and EMT in OVA-induced chronic asthma mice. Our research was the first to reveal that inhibition of LAMP2, CTSD and CTSS expression in autophagy contributed to the therapeutic efficacy of IS to asthma.
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人类巨噬细胞迁移抑制因子增强了在过敏性哮喘临床相关模型中间充质干细胞的疗效
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