Wang Yuanyuan, Zhang Nianzhi, Liu Feng, Zhou Jing, Teng Gang, Huang He
Department of Respiratory Medicine, The First Affiliated Hospital of Anhui University of Chinese Medicine, 230031 Hefei, Anhui, China.
Second Internal Medicine, Mingguang City Hospital of Traditional Chinese Medicine, 239000 Chuzhou, Anhui, China.
Discov Med. 2024 Dec;36(191):2386-2398. doi: 10.24976/Discov.Med.202436191.220.
Chronic obstructive pulmonary disease (COPD) is a prevalent yet manageable respiratory condition. However, treatments presently used normally have side effects and cannot cure COPD, making it urgent to explore effective medications. The ginsenoside Rg3 (Rg3) has been shown to have anti-inflammatory and anti-tumor properties and can improve COPD. The primary objectives of this investigation were to explore the impact of Rg3 on COPD and delve into the associated mechanisms.
models exposed human bronchial epithelial cells (BEAS-2B) to cigarette smoke extract (CSE), and models induced COPD in mice through chronic inhalation of cigarette smoke (CS). Sirtuin 1 (SIRT1) expression was regulated via cell transfection or mice infection with recombinant lentiviruses. mRNA levels were quantified using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), and SIRT protein levels were assessed by western blot or enzyme-linked immunosorbent assays (ELISA). Mitophagy was evaluated by light chain 3 (LC3) II/I and phosphatase and tensin homolog (PTEN)-induced kinase 1 (PINK1) levels, and apoptosis was determined using terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Lung function was measured with the Buxco system, and inflammation was assessed via interleukin 6 (IL-6) and keratinocyte-derived cytokine (KC) levels in bronchial alveolar lavage fluid. Lung morphological impairments were determined through Hematoxylin and Eosin (H&E) staining and mean linear intercept (MLI) measurement.
In BEAS-2B cells, CSE treatment caused a decrease in SIRT1 expression ( < 0.01) and an increase in LC3 II/I ( < 0.01) and PINK1 ( < 0.01), which were all reversed by Rg3 ( < 0.01), with 20 μM Rg3 performing the best and being used subsequently. CSE increased apoptosis of BEAS-2B cells ( < 0.01), which was reversed by Rg3 ( < 0.01). Upregulated SIRT1 further decreased levels of LC3 II/I ( < 0.001), PINK1 ( < 0.001), and cell apoptosis ( < 0.001) for CSE- and Rg3-treated cells, whereas downregulated SIRT1 reversely increased levels of LC3 II/I ( < 0.001), PINK1 ( < 0.001), and cell apoptosis ( < 0.001). The establishment of COPD caused a decrease in SIRT1 mRNA ( < 0.001), SIRT1 protein ( < 0.001), and lung functions ( < 0.001) whereas IL-6 ( < 0.001), KC ( < 0.001), lung impairment, and MLI ( < 0.001) were increased; all of these effects were reversed by Rg3 ( < 0.001). Moreover, the Rg3-induced reversion was furthered by SIRT1 upregulation ( < 0.001) and was disrupted by SIRT1 downregulation ( < 0.001).
Rg3, through activation of SIRT1, suppresses mitophagy and apoptosis, ameliorates COPD, and improves lung functions.
慢性阻塞性肺疾病（COPD）是一种常见但可控制的呼吸道疾病。然而，目前使用的治疗方法通常有副作用，且无法治愈COPD，因此迫切需要探索有效的药物。人参皂苷Rg3（Rg3）已被证明具有抗炎和抗肿瘤特性，并可改善COPD。本研究的主要目的是探讨Rg3对COPD的影响并深入研究其相关机制。
将人支气管上皮细胞（BEAS-2B）暴露于香烟烟雾提取物（CSE）中建立模型，并通过慢性吸入香烟烟雾（CS）在小鼠中诱导COPD。通过细胞转染或用重组慢病毒感染小鼠来调节沉默调节蛋白1（SIRT1）的表达。使用定量实时逆转录聚合酶链反应（qRT-PCR）定量mRNA水平，并通过蛋白质印迹或酶联免疫吸附测定（ELISA）评估SIRT蛋白水平。通过轻链3（LC3）II/I和磷酸酶和张力蛋白同源物（PTEN）诱导激酶1（PINK1）水平评估线粒体自噬，并使用末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）测定细胞凋亡。用Buxco系统测量肺功能，并通过支气管肺泡灌洗液中的白细胞介素6（IL-6）和角质形成细胞衍生细胞因子（KC）水平评估炎症。通过苏木精和伊红（H&E）染色和平均线性截距（MLI）测量确定肺形态学损伤。
在BEAS-2B细胞中，CSE处理导致SIRT1表达降低（<0.01），LC3 II/I（<0.01）和PINK1（<0.01）增加，而Rg3可逆转这些变化（<0.01），其中20μM Rg3效果最佳，随后被使用。CSE增加了BEAS-2B细胞的凋亡（<0.01），而Rg3可逆转这一现象（<0.01）。上调SIRT1进一步降低了CSE和Rg3处理细胞的LC3 II/I（<0.001）、PINK1（<0.001）水平以及细胞凋亡（<0.001），而下调SIRT1则相反地增加了LC3 II/I（<0.001）、PINK1（<0.001）水平以及细胞凋亡（<0.001）。COPD的建立导致SIRT1 mRNA（<0.001）、SIRT1蛋白（<0.001）和肺功能（<0.001）降低，而IL-6（<0.001）、KC（<0.001）、肺损伤和MLI（<0.001）增加；所有这些影响均被Rg3逆转（<0.001）。此外，Rg3诱导的逆转作用通过上调SIRT1进一步增强（<0.001），而被下调SIRT1破坏（<0.001）。
Rg3通过激活SIRT1，抑制线粒体自噬和细胞凋亡，改善COPD并提高肺功能。
