Zhang Lili, Qu Sihao, Wang Lu, Wang Chunguo, Yu Qinghe, Zhang Zhimin, Diao Yirui, Zhang Binbin, Li Yadong, Shi Yuanyuan, Wang Peng
School of Life Sciences, Beijing University of Chinese Medicine, Beijing, China.
School of Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China.
Front Pharmacol. 2021 Apr 22;12:661129. doi: 10.3389/fphar.2021.661129. eCollection 2021.
Pulmonary Fibrosis (PF) is an interstitial lung disease characterized by excessive accumulation of extracellular matrix in the lungs, which disrupts the structure and gas exchange of the alveoli. There are only two approved therapies for PF, nintedanib (Nib) and pirfenidone. Therefore, the use of Chinese medicine for PF is attracting attention. Tianlongkechuanling (TL) is an effective Chinese formula that has been applied clinically to alleviate PF, which can enhance lung function and quality of life. The potential effects and specific mechanisms of TL have not been fully explored, yet. In the present study, proteomics was performed to explore the therapeutic protein targets of TL on Bleomycin (BLM)-induced Pulmonary Fibrosis. BLM-induced PF mice models were established. Hematoxylineosin staining and Masson staining were used to analyze histopathological changes and collagen deposition. To screen the differential proteins expression between the Control, BLM, BLM + TL and BLM + Nib (BLM + nintedanib) groups, quantitative proteomics was performed using tandem mass tag (TMT) labeling with nanoLC-MS/MS [nano liquid chromatographymass spectrometry]). Changes in the profiles of the expressed proteins were analyzed using the bioinformatics tools Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). The protein-protein interactions (PPI) were established by STRING. Expressions of α-smooth muscle actin (α-SMA), (), () and enzymes in arginase-ornithine pathway were detected by Western blot or RT-PCR. TL treatments significantly ameliorated BLM-induced collagen deposition in lung tissues. Moreover, TL can inhibit the protein expressions of α-SMA and the mRNA expressions of and . Using TMT technology, we observed 253 differentially expressed proteins related to PPI networks and involved different KEGG pathways. Arginase-ornithine pathway is highly significant. The expression of Arg was significantly decreased after TL treatments. Administration of TL in BLM-induced mice resulted in decreasing pulmonary fibrosis. Our findings propose that the down regulation of arginase-ornithine pathway expression with the reduction of arginase biosynthesis is a central mechanism and potential treatment for pulmonary fibrosis with the prevention of TL.
肺纤维化（PF）是一种间质性肺疾病，其特征是肺中细胞外基质过度积累，这会破坏肺泡的结构和气体交换。目前仅有两种获批用于治疗PF的药物，即尼达尼布（Nib）和吡非尼酮。因此，中医药治疗PF正受到关注。天龙咳喘灵（TL）是一种有效的中药方剂，已在临床上应用于缓解PF，可增强肺功能和提高生活质量。然而，TL的潜在作用和具体机制尚未得到充分探索。在本研究中，采用蛋白质组学方法探索TL对博莱霉素（BLM）诱导的肺纤维化的治疗性蛋白质靶点。建立了BLM诱导的PF小鼠模型。采用苏木精-伊红染色和Masson染色分析组织病理学变化和胶原沉积。为筛选对照组、BLM组、BLM+TL组和BLM+Nib（BLM+尼达尼布）组之间差异蛋白质表达，使用串联质谱标签（TMT）标记结合纳升液相色谱-质谱联用（nanoLC-MS/MS）进行定量蛋白质组学分析。使用基因本体论（GO）和京都基因与基因组百科全书（KEGG）生物信息学工具分析表达蛋白质谱的变化。通过STRING建立蛋白质-蛋白质相互作用（PPI）。通过蛋白质免疫印迹法或逆转录-聚合酶链反应检测α-平滑肌肌动蛋白（α-SMA）、（）、（）和精氨酸酶-鸟氨酸途径中酶的表达。TL治疗显著改善了BLM诱导的肺组织胶原沉积。此外，TL可抑制α-SMA的蛋白质表达以及（）和（）的mRNA表达。使用TMT技术，我们观察到253种与PPI网络相关且涉及不同KEGG途径的差异表达蛋白质。精氨酸酶-鸟氨酸途径高度显著。TL治疗后精氨酸酶（Arg）的表达显著降低。在BLM诱导的小鼠中给予TL导致肺纤维化减轻。我们的研究结果表明，随着精氨酸酶生物合成减少，精氨酸酶-鸟氨酸途径表达下调是肺纤维化的核心机制以及TL预防肺纤维化的潜在治疗方法。
