Jiao Pengfei, Wang Yingrui, Sang Tianqing, Jiao Jing, Li Yameng
Department of Respiration and Intensive Care Unit, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Department of Hematology Oncology, The First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, China.
Ann Transl Med. 2022 Jun;10(12):707. doi: 10.21037/atm-22-2629.
The aim of this study was to discover the molecular mechanism of betulin palliative therapy for chronic obstructive pulmonary disease (COPD) based on the receptor target of gated ion channel.
A COPD mouse model was developed. Changes in pulmonary ventilation function, pulmonary airway and vascular remodeling indicators, inflammatory cells, and inflammatory factors were determined after betulin intervention, and the pathological alterations of lung tissues were detected. An in vitro experimental model was constructed to observe the influence of betulin at varying concentrations on the proliferation of human bronchial epidermal cell line (16-HBE) cells and changes in inflammatory factors in cell supernatant. The expression levels of key proteins in 16-HBE cells transfected with overexpressed or silenced genes were determined through quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot.
After betulin intervention, pulmonary ventilation function in the 20 mg/kg betulin and 40 mg/kg betulin groups was improved. Levels of white blood cells (WBCs), neutrophils (Ns), tumor necrosis factor (TNF), TNF-ɑ, interleukin (IL)-1β, and IL-6 in the 2 groups also decreased significantly (all P<0.05). The pathological changes in COPD mice were detected. After betulin intervention, the pathological injury of the lung was reduced, the pathological score decreased significantly, and the remodeling indicators of pulmonary airway and pulmonary vessels diminished remarkably (all P<0.05). Betulin had no effect on the proliferation of 16-HBE cells in vitro. After cigarette smoke extract (CSE) stimulation, the rate of survival for 16-HBE cells decreased significantly. After betulin treatment, the survival rate of 16-HBE cells augmented remarkably, and the levels of TNF-ɑ, IL-6, and IL-1β in cell supernatant reduced substantially (all P<0.05). 16-HBE with overexpression and knockdown of was constructed. After being treated with betulin, the relative expression levels of messenger RNA (mRNA) of ERK, JNK, rho-associated protein kinase (ROCK), nuclear factor-κB (NF-κB), and p38 in 16-HBE cells with overexpression or knockdown were decreased significantly (all P<0.05), but the above indicators were largely unchanged (all P>0.05).
Betulin relieved lung pathological injury, ameliorated lung ventilation function, and diminished the level of inflammatory factors in COPD mice, playing a therapeutic role via the signaling pathway.
本研究旨在基于门控离子通道的受体靶点，探寻桦木醇对慢性阻塞性肺疾病（COPD）进行姑息治疗的分子机制。
建立COPD小鼠模型。在桦木醇干预后，测定肺通气功能、肺气道和血管重塑指标、炎症细胞及炎症因子的变化，并检测肺组织的病理改变。构建体外实验模型，观察不同浓度桦木醇对人支气管表皮细胞系（16-HBE）细胞增殖及细胞上清液中炎症因子变化的影响。通过定量逆转录聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测过表达或沉默基因的16-HBE细胞中关键蛋白的表达水平。
桦木醇干预后，20mg/kg桦木醇组和40mg/kg桦木醇组的肺通气功能得到改善。两组的白细胞（WBC）、中性粒细胞（N）、肿瘤坏死因子（TNF）、TNF-α、白细胞介素（IL）-1β和IL-6水平也显著降低（均P<0.05）。检测了COPD小鼠的病理变化。桦木醇干预后，肺组织病理损伤减轻，病理评分显著降低，肺气道和肺血管的重塑指标明显减少（均P<0.05）。桦木醇对体外16-HBE细胞的增殖无影响。香烟烟雾提取物（CSE）刺激后，16-HBE细胞的存活率显著降低。桦木醇处理后，16-HBE细胞的存活率显著提高，细胞上清液中TNF-α、IL-6和IL-1β水平大幅降低（均P<0.05）。构建了过表达和敲低的16-HBE细胞。用桦木醇处理后，过表达或敲低的16-HBE细胞中细胞外调节蛋白激酶（ERK）、c-Jun氨基末端激酶（JNK）、rho相关蛋白激酶（ROCK）、核因子-κB（NF-κB）和p38的信使核糖核酸（mRNA）相对表达水平显著降低（均P<0.05），但上述指标基本无变化（均P>0.05）。
桦木醇减轻了COPD小鼠的肺病理损伤，改善了肺通气功能，降低了炎症因子水平，通过信号通路发挥治疗作用。
