Cigarette smoke extracts induce apoptosis in Raw264.7 cells via endoplasmic reticulum stress and the intracellular Ca2+/P38/STAT1 pathway

Cigarette smoke (CS) exposure is a risk factor for chronic obstructive pulmonary disease (COPD). CS exposure impairs the ability of killing pathogens in macrophages, which might be due to the abnormal apoptosis induced by CS. This study explored the effects and mechanisms of cigarette smoke extract (CSE) on the apoptosis of macrophages in vitro. Raw264.7 cells were treated with CSE at different concentrations, and viability and apoptosis of cells was accessed. The protein expression was detected by western blot. The intracellular Ca2+ level was evaluated by Fluo-4 AM probe assay. CSE induced the apoptosis and increased the expression of cleaved caspase 3, which were attenuated by a caspase inhibitor. CSE increased the expression of CHOP, BiP and P-eif2 alpha, and the inhibitor of endoplasmic reticulum stress (ERS) decreased the apoptosis induced by CSE. Phosphorylation levels of P38, JNK and ERK1/2 were increased following incubation with CSE. Only P38 inhibitor significantly reduced apoptosis induced by CSE, while ERK1/2 inhibitor promoted apoptosis. Phosphorylation of STAT1 at Ser727 was activated by CSE and attenuated by the P38 inhibitor. Finally, CSE increased the level of intracellular Ca2+, and calcium chelator partly attenuated the apoptosis and phosphorylation of P38 and STAT1 induced by CSE. CSE induced a caspase 3-dependent apoptosis in Raw264.7 cells via ERS and intracellular Ca2+/P38/STAT1 pathway.

Automated summary

Cigarette smoke extract induces apoptosis in Raw264.7 cells through endoplasmic reticulum stress and the intracellular Ca2+/P38/STAT1 pathway. The increased expression of CHOP, BiP, and P-eif2 alpha were observed, with P38 inhibitor significantly reducing apoptosis and ERK1/2 inhibitor promoting apoptosis. This study reveals the mechanisms by which cigarette smoke leads to macrophage apoptosis.