Supplementary Material for: Anti-Inflammatory Effects of Japanese Herbal Medicine Hochuekkito in a Mouse Model of Acute Exacerbation of Chronic Obstructive Pulmonary Disease

<b><i>Introduction:</i></b> The traditional Japanese herbal medicine hochuekkito (TJ-41) has been reported to ameliorate systemic inflammation and malnutrition in patients with chronic obstructive pulmonary disease (COPD). TJ-41 has also been known to have preventive effects against influenza virus infection. However, its role in the acute exacerbation of COPD (AECOPD) remains to be elucidated. Our previous study established a murine model of viral infection-associated AECOPD that was induced by intratracheal administration of porcine pancreatic elastase (PPE) and polyinosinic-polycytidylic acid [poly(I:C)]. Here, we used this model and investigated the effects of TJ-41 in AECOPD. <b><i>Methods:</i></b> Specific pathogen-free C57BL/6J mice were used. A COPD model was induced by treating mice intratracheally with PPE on day 0. To generate the murine model of AECOPD, poly(I:C) was administered intratracheally following PPE treatment on days 22–24. Mice were sacrificed and analyzed on day 25. Mice were fed a diet containing 2% TJ-41 or a control diet. <b><i>Results:</i></b> Daily oral intake of TJ-41 significantly decreased the numbers of neutrophils and lymphocytes in the bronchoalveolar lavage fluid (BALF), which was accompanied by decreased transcripts of CXC chemokines involved in neutrophil migration, viz., <i>Cxcl1</i> and <i>Cxcl2,</i> in whole lung homogenates and reduced Cxcl2 concentration in BALF. <b><i>Conclusion:</i></b> This study demonstrates the anti-inflammatory effects of TJ-41 in a mouse model of AECOPD, suggesting the effectiveness of TJ-41 for the management of COPD. Clinical investigations evaluating the therapeutic efficacy of TJ-41 in AECOPD would be meaningful.

**引言：** 传统日本汉方药防风通圣散（hochuekkito，TJ-41）据报道可改善慢性阻塞性肺疾病（chronic obstructive pulmonary disease, COPD）患者的全身炎症与营养不良状态。此外，TJ-41对流感病毒感染具有预防作用，但其在慢性阻塞性肺疾病急性加重期（acute exacerbation of COPD, AECOPD）中的作用仍有待阐明。本团队前期已构建病毒感染相关AECOPD小鼠模型，该模型通过气管内给予猪胰弹性蛋白酶（porcine pancreatic elastase, PPE）与聚肌胞苷酸[poly(I:C)]诱导得到。本研究利用该模型，探究TJ-41在AECOPD中的作用效果。
**方法：** 实验采用无特定病原体（specific pathogen-free, SPF）级C57BL/6J小鼠。于第0天经气管内给予PPE构建COPD小鼠模型。为制备AECOPD小鼠模型，于PPE给药后的第22~24天，经气管内给予poly(I:C)。于第25天处死小鼠并开展相关分析。小鼠喂食含2% TJ-41的饲料或普通对照饲料。
**结果：** 每日经口摄入TJ-41可显著降低支气管肺泡灌洗液（bronchoalveolar lavage fluid, BALF）中的中性粒细胞与淋巴细胞数量，同时可下调全肺组织匀浆中参与中性粒细胞迁移的CXC趋化因子（即Cxcl1与Cxcl2）的转录水平，并降低BALF中Cxcl2的浓度。
**结论：** 本研究证实了TJ-41在AECOPD小鼠模型中的抗炎作用，提示其可用于慢性阻塞性肺疾病的临床管理。开展评估TJ-41对AECOPD治疗效果的临床研究具有重要意义。