Wu Xin-Cheng, Liu Yu, Bai Zheng-Ping
Hunan University of Chinese Medicine Changsha 410208, China Department of Pulmonary and Critical Care Medicine,Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine Nanning 530011, China.
the Affilited Hospital of Hunan Academy of Traditional Chinese Medicine Changsha 410006, China.
Zhongguo Zhong Yao Za Zhi. 2023 Jun;48(11):3055-3065. doi: 10.19540/j.cnki.cjcmm.20221014.702.
This paper aimed to investigate the effects of high mobility group box 1(HMGB1)-mediated pulmonary artery smooth muscle cell pyroptosis and immune imbalance on chronic obstructive pulmonary disease-associated pulmonary hypertension(COPD-PH) in rats and the intervening mechanism of Compound Tinglizi Decoction. Ninety rats were randomly divided into a normal group, a model group, low-dose, medium-dose, and high-dose Compound Tinglizi Decoction groups, and a simvastatin group. The rat model of COPD-PH was established by fumigation combined with lipopolysaccharide(LPS) intravascular infusion, which lasted 60 days. Rats in the low, medium, and high-dose Compound Tinglizi Decoction groups were given 4.93, 9.87, and 19.74 g·kg(-1) Compound Tinglizi Decoction by gavage, respectively. Rats in the simvastatin group were given 1.50 mg·kg(-1) simvastatin by gavage. After 14 days, the lung function, mean pulmonary artery pressure, and arterial blood gas of rats were analyzed. Lung tissues of rats were collected for hematoxylin-eosin(HE) staining to observe the pathological changes. Real-time fluorescent quantitative polymerase chain reaction(qRT-PCR) was used to determine the expression of related mRNA in lung tissues, Western blot(WB) was used to determine the expression of related proteins in lung tissues, and enzyme linked immunosorbent assay(ELISA) was used to determine the levels of inflammatory factors in the lung tissues of rats. The ultrastructure of lung cells was observed by transmission electron microscope. The forced vital capacity(FVC), forced expiratory volume in 0.3 second(FEV_(0.3)), FEV_(0.3)/FVC, peek expiratory flow(PEF), respiratory dynamic compliance(Cdyn), arterial partial pressure of oxygen(PaO_2), and arterial oxygen saturation(SaO_2) were increased, and resistance of expiration(Re), mean pulmonary arterial pressure(mPAP), right ventricular hypertrophy index(RVHI), and arterial partial pressure of carbon dioxide(PaCO_2) were decreased by Compound Tinglizi Decoction in rats with COPD-PH. Compound Tinglizi Decoction inhibited the protein expression of HMGB1, receptor for advanced glycation end products(RAGE), pro caspase-8, cleaved caspase-8, and gasdermin D(GSDMD) in lung tissues of rats with COPD-PH, as well as the mRNA expression of HMGB1, RAGE, and caspase-8. Pulmonary artery smooth muscle cell pyroptosis was inhibited by Compound Tinglizi Decoction. Interferon-γ(IFN-γ) and interleukin-17(IL-17) were reduced, and interleukin-4(IL-4) and interleukin-10(IL-10) were incresead by Compound Tinglizi Decoction in lung tissues of rats with COPD-PH. In addition, the lesion degree of trachea, alveoli, and pulmonary artery in lung tissues of rats with COPD-PH was improved by Compound Tinglizi Decoction. Compound Tinglizi Decoction had dose-dependent effects. The lung function, pulmonary artery pressure, arterial blood gas, inflammation, trachea, alveoli, and pulmonary artery disease have been improved by Compound Tinglizi Decoction, and its mechanism is related to HMGB1-mediated pulmonary artery smooth muscle cell pyroptosis and helper T cell 1(Th1)/helper T cell 2(Th2), helper T cell 17(Th17)/regulatory T cell(Treg) imbalance.
本文旨在探讨高迁移率族蛋白B1（HMGB1）介导的肺动脉平滑肌细胞焦亡及免疫失衡对大鼠慢性阻塞性肺疾病相关性肺动脉高压（COPD-PH）的影响以及复方葶苈子汤的干预机制。将90只大鼠随机分为正常组、模型组、复方葶苈子汤低剂量组、中剂量组、高剂量组和辛伐他汀组。采用烟熏联合脂多糖（LPS）血管内注射法建立COPD-PH大鼠模型，持续60天。复方葶苈子汤低、中、高剂量组大鼠分别按4.93、9.87、19.74 g·kg⁻¹灌胃给予复方葶苈子汤。辛伐他汀组大鼠按1.50 mg·kg⁻¹灌胃给予辛伐他汀。14天后，分析大鼠肺功能、平均肺动脉压及动脉血气。采集大鼠肺组织进行苏木精-伊红（HE）染色观察病理变化。采用实时荧光定量聚合酶链反应（qRT-PCR）检测肺组织中相关mRNA表达，蛋白质免疫印迹法（WB）检测肺组织中相关蛋白表达，酶联免疫吸附测定（ELISA）检测大鼠肺组织中炎症因子水平。通过透射电子显微镜观察肺细胞超微结构。复方葶苈子汤可使COPD-PH大鼠用力肺活量（FVC）、0.3秒用力呼气量（FEV₀.₃）、FEV₀.₃/FVC、呼气峰流速（PEF）、呼吸动态顺应性（Cdyn）、动脉血氧分压（PaO₂）及动脉血氧饱和度（SaO₂）升高，呼气阻力（Re）、平均肺动脉压（mPAP）、右心室肥厚指数（RVHI）及动脉血二氧化碳分压（PaCO₂）降低。复方葶苈子汤可抑制COPD-PH大鼠肺组织中HMGB1、晚期糖基化终末产物受体（RAGE）、前半胱天冬酶-8、裂解的半胱天冬酶-8及gasdermin D（GSDMD）蛋白表达，以及HMGB1、RAGE及半胱天冬酶-8的mRNA表达。复方葶苈子汤可抑制肺动脉平滑肌细胞焦亡。复方葶苈子汤可使COPD-PH大鼠肺组织中干扰素-γ（IFN-γ）及白细胞介素-17（IL-17）减少，白细胞介素-4（IL-4）及白细胞介素-10（IL-10）增加。此外，复方葶苈子汤可改善COPD-PH大鼠肺组织气管、肺泡及肺动脉病变程度。复方葶苈子汤具有剂量依赖性作用。复方葶苈子汤可改善肺功能、肺动脉压、动脉血气、炎症、气管、肺泡及肺动脉病变状况，其机制与HMGB1介导的肺动脉平滑肌细胞焦亡及辅助性T细胞1（Th1）/辅助性T细胞2（Th2）、辅助性T细胞17（Th17）/调节性T细胞（Treg）失衡有关。
