Ding Shumin, Hou Xuefeng, Wang Gang, Qiu Huihui, Liu Ying, Zhou Yuanli, Du Mei, Tan Xiaobin, Song Jie, Wei Yingjie, Shu Luan, Li Zhiyong, Feng Liang, Jia Xiaobin
School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, China.
Key Laboratory of New Drug Delivery System of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Nanjing, China.
Front Pharmacol. 2018 Feb 20;9:107. doi: 10.3389/fphar.2018.00107. eCollection 2018.
Excessive autophagy plays a crucial role in cigarette smoking extract (CSE)-induced inflammation response and oxidative damage of respiratory epithelial cells. The components from L. (CCE) have been shown to be beneficial for CSE-induced epithelial cells injury. However, whether its protection on CSE-stress injury is related to its regulation on autophagy remains still unclear. In this study, CCE, containing mainly wedelolactone of 45.88% and demethylwedelolactone of 23.74%, could improve significantly 10%CSE-induced cell viability of normal human bronchial epithelial (NHBE) cells using CCK-8 kit. We revealed that CCE could remarkably increase autophagic factors Beclin-1, Atg5, ATF4 proteins expression levels and the transformation of LC3-I to LC3-II. Additionally, CCE up-regulated significantly p-p16 and p-p21 phosphorylation levels whereas down-regulated p-p53 in NHBE cells. The changes of typical autolysosom and representative autophagosome in the presence of CCE or/and autophagy inhibitor chloroquine (CQ) were also observed by transmission electron microscopy. These data demonstrated that CCE reduced CSE-induced autophagy flux activation in NHBE cells. The blockade of CCE on autophagy flux contributes to its protection against CSE-induced NHBE cells damage, and CCE is promising to be combination therapeutic molecules to excessive autophagic damage in respiratory diseases.
过度自噬在香烟烟雾提取物（CSE）诱导的呼吸道上皮细胞炎症反应和氧化损伤中起关键作用。已证明来自[植物名称未给出]（CCE）的成分对CSE诱导的上皮细胞损伤有益。然而，其对CSE应激损伤的保护作用是否与其对自噬的调节有关仍不清楚。在本研究中，主要含有45.88%的蟛蜞菊内酯和23.74%的去甲基蟛蜞菊内酯的CCE，使用CCK-8试剂盒可显著提高10% CSE诱导的正常人支气管上皮（NHBE）细胞的活力。我们发现CCE可显著增加自噬因子Beclin-1、Atg5、ATF4蛋白的表达水平以及LC3-I向LC3-II的转化。此外，CCE显著上调NHBE细胞中p-p16和p-p21的磷酸化水平，而下调p-p53。还通过透射电子显微镜观察了在存在CCE或/和自噬抑制剂氯喹（CQ）的情况下典型自溶酶体和代表性自噬体的变化。这些数据表明CCE降低了CSE诱导的NHBE细胞自噬流激活。CCE对自噬流的阻断有助于其对CSE诱导的NHBE细胞损伤的保护作用，并且CCE有望成为治疗呼吸系统疾病中过度自噬损伤的联合治疗分子。
