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Effect of CSE on M1/M2 polarization in alveolar and peritoneal macrophages at different concentrations and exposure in vitro

Author:
Feng, Haoshen, Yin, Yan, Ren, Yuan, Li, Menglu, Zhang, Dan, Xu, Mingtao, Cai, Xu, Kang, Jian
Source:
In vitro cellular & developmental biology 2020 v.56 no.2 pp. 154-164
ISSN:
1071-2690
Subject:
cell viability, cigarettes, disease progression, exposure duration, flow cytometry, gene expression, inducible nitric oxide synthase, interleukin-1beta, macrophages, messenger RNA, proteins, quantitative polymerase chain reaction, rats, respiratory tract diseases, smoke, smoking (habit), transforming growth factor beta 1, tumor necrosis factor-alpha
Abstract:
Cigarette smoke exposure is one of the main etiologies for chronic obstructive pulmonary disease. Moreover, cigarette smoke participates in disease progression by inducing abnormal macrophage polarization; however, the effects of cigarette smoke on M1/M2 macrophage polarization have not been established. The aim of the current study was to determine the effects of cigarette smoke extract (CSE) on M1/M2 macrophage polarization in alveolar and peritoneal macrophages (AM and PM, respectively) at different concentrations and exposure times. Rat AM and PM were cultured with CSE at different concentrations. CCK-8 was used as an indicator of cell viability, and mRNA expression of M1 (iNOS, TNF-α, and IL-1β) and M2 markers (arg-1, CD206, and TGF-β1) were measured at 3, 6, 9, 12, and 24 h using qPCR. Expressions of CD86 and CD206 proteins at 12 h were determined using flow cytometry, and the iNOS/arg-1 ratio was used to determine the polarization dominance of M1 and M2. M2 subtypes were detected at 12 h using qPCR and flow cytometry. CSE increased the expression of iNOS, TNF-α, and IL-1β mRNA, and the proportion of CD86-positive cells in AM and PM promoted M1 polarization, and M1 polarization was continuously enhanced as exposure time and concentration increased. CSE reduced the expression of arg-1, CD206, and TGF-β1 mRNA and the proportion of CD206-positive cells in AM and PM and inhibited M2 polarization. At 9–24 h of CSE exposure, the expression of arg-1 in AM and PM gradually increased, showing tendency towards activation of M2 polarization. Besides, CSE might induce M2b and M2d polarization at 12 h. After 12 h of CSE exposure, transformation from M1 to M2 polarization dominance was shown in AM; however, M1 polarization was continuously enhanced in PM within 24 h of CSE exposure. CSE promoted M1 polarization in macrophages, exhibiting dynamic regulatory effects on M2 polarization, first as a suppressor and then as a promoter. The polarization change induced by CSE on AM was more sensitive than PM.
Agid:
6856803