Resveratrol Protects Against Nicotine-Induced Apoptosis by Enhancing Autophagy in BEAS-2B Lung Epithelial Cells

Abstract

Background: Nicotine (Nic), the major component of tobacco products, can induce apoptosis in lung epithelial cells, and the resulting damage contributes to chronic obstructive pulmonary disease. Apoptosis is closely related to autophagy. Resveratrol (Res) can induce autophagy and inhibit apoptosis. Therefore, the present study investigated whether Nic induces apoptosis of lung epithelial cells by regulating autophagy and the effect of Res on apoptosis of Nic-exposed lung epithelial cells. Methods: The BEAS-2B lung epithelial cell line was used to study the harmful effects of Nic and the potential benefits of Res as well as the underlying mechanisms. Viability and apoptosis were examined using the Cell Counting Kit-8 and annexin V-propidium iodide staining, respectively. The expression of levels of apoptosis-related proteins, autophagy-related proteins, and members of the PI3K/Akt/mTOR pathway was measured by western blotting. Autophagic flux was detected via mRFP-GFP-LC3 double-labeled adenovirus transfection and transmission electron microscopy. Results: Nic significantly reduce the viability and increased the apoptosis of BEAS-2B cells in a concentration-dependent manner. Nic treatment also increased the numbers of autophagosomes in BEAS-2B cells and upregulated LC3II and p62 expression. Moreover, Res at concentration of 2, 10, and 50 μM protected BEAS-2B cells from Nic apoptosis, and the expression of LC3II increased further and p62 decreased in Res pretreatment group. Apart from this, Res reduced Akt and mTOR phosphorylation. Subsequently, upon inhibiting PI3K phosphorylation by PI3K inhibitors, BEAS-2B cell autophagy induced by Res was obviously abolished. Conclusions: Nic-induced BEAS-2B cell apoptosis by inhibiting the late-stage autophagic flux, but Res could protect BEAS-2B cells from the detrimental effects of nicotine by enhancing autophagy via the PI3K/Akt/mTOR pathway. These results will provide an experimental basis for the prevention and treatment of COPD.