Lu Feng, Yang Han, Lin Si-Ding, Zhao Li, Jiang Chang, Chen Zhi-Bin, Liu Ying-Ying, Kan Yong-Jun, Hu Juan, Pang Wen-Sheng
School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou, China.
Respiratory Department, The Second Affiliated Hospital of Fujian Traditional Chinese Medical University, Fuzhou, China.
Front Pharmacol. 2020 Jun 9;11:850. doi: 10.3389/fphar.2020.00850. eCollection 2020.
We have explored the method of extraction and purification of cyclic-peptide extract (CPE) from (Miq.) Pax. (Taizishen, TZS), characterized the structure about cyclic-peptide compounds and investigated the biological activity of CPE attenuating chronic obstructive pulmonary disease (COPD) in rats. The CPE from TZS was obtained by ethyl acetate, petroleum ether, hot water extraction, and alcohol-precipitation. Cyclic-peptide structures were distinguished using ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). Rats were induced by solid combustibles smoke (SCS) for the COPD model, and the anti-COPD activity of CPE was detected using lung airway resistance and dynamic lung compliance, as well as pulmonary tissue hematoxylin and eosin (HE) staining. The relevant inflammatory cytokines were assayed by enzyme-linked immunosorbent assay (ELISA). CPE obtained from TZS contained 12 cyclic-peptide constituents; the purity was up to 92.94%. CPE (200, 400, or 500 mg/kg/day) was given to SCS-induced COPD model rats orally for 15 days. The results showed that in rats given CPE (400 mg/kg/day) there was a sharp fall in lung airway resistance but a rise in dynamic lung compliance. The image analysis of lung tissue sections suggested that CPE could decrease the degree of alveolar destruction (0.05), alleviate lung inflammation, increase alveolar space, and improve the infiltration of inflammatory cells. CPE was found to reduce the levels of TNF-α, but increase IL-10, adjusting multiple cytokines in rat serum; the TLR4 mRNA, MyD88 mRNA and AP-1 mRNA levels, the expressing levels of p-JNK, p-p38 and p-TAK1 protein were significantly down regulated in rat alveolar macrophages. CPE intervention could improve the pulmonary ventilation function on COPD rats, which may be related to its effect in inhibiting the abnormal activation of the TLR4-MyD88-JNK/p38 pathway. This is the first report that the CPE of TZS lessens the severity of COPD episodes. The new preparation process of CPEs implements the anticipated goal, which is to refine CPE and actualize quality control.
我们探索了从太子参（Taizishen，TZS）中提取和纯化环肽提取物（CPE）的方法，对环肽化合物的结构进行了表征，并研究了CPE减轻大鼠慢性阻塞性肺疾病（COPD）的生物活性。通过乙酸乙酯、石油醚、热水提取和醇沉法从太子参中获得CPE。使用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q-TOF-MS/MS）鉴别环肽结构。用固体可燃物烟雾（SCS）诱导大鼠建立COPD模型，通过肺气道阻力、动态肺顺应性以及肺组织苏木精和伊红（HE）染色检测CPE的抗COPD活性。采用酶联免疫吸附测定（ELISA）法检测相关炎性细胞因子。从太子参中获得的CPE含有12种环肽成分，纯度高达92.94%。将CPE（200、400或500mg/kg/天）口服给予SCS诱导的COPD模型大鼠，持续15天。结果显示，给予CPE（400mg/kg/天）的大鼠肺气道阻力急剧下降，动态肺顺应性升高。肺组织切片的图像分析表明，CPE可降低肺泡破坏程度（P<0.05），减轻肺部炎症；增加肺泡间隙，改善炎性细胞浸润。发现CPE可降低TNF-α水平，但增加IL-10水平，调节大鼠血清中的多种细胞因子；大鼠肺泡巨噬细胞中TLR4 mRNA、MyD88 mRNA和AP-1 mRNA水平以及p-JNK、p-p38和p-TAK1蛋白的表达水平均显著下调。CPE干预可改善COPD大鼠的肺通气功能，这可能与其抑制TLR4-MyD88-JNK/p38通路异常激活的作用有关。这是关于太子参CPE减轻COPD发作严重程度的首次报道。CPE的新制备工艺实现了预期目标，即精制CPE并实现质量控制。
