Abstract
Ophiocordyceps sinensis (O. sinensis) seems to be able to alleviate airway epithelial cell senescence in chronic obstructive pulmonary disease (COPD). The objective of the study is to evaluate the effect of O. sinensis on airway epithelial senescence in the COPD model both in vitro and in vivo. We observed the expression of P16 and P21 in the airway epithelia of 30 patients with COPD. The optimal concentration of O. sinensis and exposure time of the cigarette smoke extract (CSE) were determined in vitro, and senescence-associated β-galactosidase (SA-β-gal) and 5-bromodeoxyuridine (BrdU) were used to evaluate the senescence and proliferation of human bronchial epithelial (16HBE) cells pretreated with O. sinensis by staining kits. COPD model rats were treated with O. sinensis at various concentrations to determine the changes in P16 and P21 expression in airway epithelial tissues. It was found that the expression levels of P16 and P21 were higher in the airway epithelia of COPD patients than those in the control group based on immunohistochemical staining, real-time quantitative PCR, and western blotting. The CSE could induce 16HBE cell senescence, and O. sinensis could alleviate CSE-induced senescence and promote the proliferation of 16HBE cells. The expression levels of P16 and P21 were also higher in the airway epithelia of COPD model rats; however, the levels of P16 and P21 in the groups treated with all concentrations of O. sinensis were obviously lower than those in the COPD model group based on real-time quantitative PCR and western blotting. In conclusion, the CSE can induce airway epithelium senescence, and O. sinensis can inhibit CSE-induced cellular senescence, both in vitro and in vivo.
Highlights
Chronic obstructive pulmonary disease (COPD) is characterized by an expiratory airflow limitation that is not fully reversible and is associated with an anomalous inflammatory reaction, mainly in response to tobacco smoke
All biopsy tissues were obtained from the Qilu Hospital of Shandong University (Jinan, Shandong, China) between September 2013 and December 2015. irty COPD specimens were obtained from lung cancer resections, and nontumor lung tissues were collected when patients were diagnosed with COPD. irty normal lung tissues were obtained from body donations because of trauma or death; these individuals had no obvious pulmonary disease, and the samples were used as a control group
For groups treated with different O. sinensis concentrations, the protein and mRNA expression levels of P16 and P21 were remarkably lower in rats treated with a moderate dose and high dose of O. sinensis than those in rats treated with a low dose of O. sinensis (Figures 3(a)–3(e)). ese results indicate that O. sinensis could inhibit the expression of P16 and P21 in the airway epithelial cells of COPD rats, indicating that O. sinensis may play a protective role in airway epithelial cellular senescence
Summary
Chronic obstructive pulmonary disease (COPD) is characterized by an expiratory airflow limitation that is not fully reversible and is associated with an anomalous inflammatory reaction, mainly in response to tobacco smoke. Previous reports have demonstrated the critical role of cellular senescence in respiratory diseases, such as asthma, pulmonary fibrosis, and lung cancer, especially in COPD [3,4,5]. Cellular senescence affects regeneration ability by reducing the amount of hematopoietic stem cells, and aging cells release cytokines to further enhance the inflammatory response, which leads to airway inflammation and airway remodeling in COPD patients [8, 9]. The effects of O. sinensis on cell senescence in COPD have not been reported its functional roles have been widely documented. Based on these previous studies, we hypothesize that the cigarette smoke extract (CSE) can induce airway epithelial senescence in COPD, and O. sinensis can alleviate CSE-induced cellular senescence. Based on these previous studies, we hypothesize that the cigarette smoke extract (CSE) can induce airway epithelial senescence in COPD, and O. sinensis can alleviate CSE-induced cellular senescence. erefore, this study investigates how O. sinensis regulates cellular senescence in COPD airway epithelium in vivo and in vitro
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