The proliferation, migration, cytokine release, and contraction of airway smooth muscle cells are key events in the airway remodeling process that occur in lung disease such as asthma, chronic obstruction pulmonary disease, and cancer. These events can be modulated by a number of factors, including ...
The proliferation, migration, cytokine release, and contraction of airway smooth muscle cells are key events in the airway remodeling process that occur in lung disease such as asthma, chronic obstruction pulmonary disease, and cancer. These events can be modulated by a number of factors, including cigarette smoke extract (CSE). CSE-induced alterations in the viability, migration, and contractile abilities of normal human airway cells remain unclear. This study investigated the effect of CSE on cell viability, migration, tumor necrosis factor (TNF)-${\alpha}$ secretion, and contraction in normal human bronchial smooth muscle cells (HBSMCs). Treatment of HBSMCs with 10% CSE induced cell death, and the death was accompanied by the generation of reactive oxygen species (ROS). CSE-induced cell death was reduced by N-acetyl-l-cysteine (NAC), an ROS scavenger. In addition, CSE reduced the migration ability of HBSMCs by 75%. The combination of NAC with CSE blocked the CSE-induced reduction of cell migration. However, CSE had no effect on TNF-${\alpha}$ secretion and NF-${\kappa}B$ activation. CSE induced an increase in intracellular $Ca^{2+}$ concentration in 64% of HBSMCs. CSE reduced the contractile ability of HBSMCs, and the ability was enhanced by NAC treatment. These results demonstrate that CSE treatment induces cell death and reduces migration and contraction by increasing ROS generation in normal HBSMCs. These results suggest that CSE may induce airway change through cell death and reduction in migration and contraction of normal HBSMCs.
The proliferation, migration, cytokine release, and contraction of airway smooth muscle cells are key events in the airway remodeling process that occur in lung disease such as asthma, chronic obstruction pulmonary disease, and cancer. These events can be modulated by a number of factors, including cigarette smoke extract (CSE). CSE-induced alterations in the viability, migration, and contractile abilities of normal human airway cells remain unclear. This study investigated the effect of CSE on cell viability, migration, tumor necrosis factor (TNF)-${\alpha}$ secretion, and contraction in normal human bronchial smooth muscle cells (HBSMCs). Treatment of HBSMCs with 10% CSE induced cell death, and the death was accompanied by the generation of reactive oxygen species (ROS). CSE-induced cell death was reduced by N-acetyl-l-cysteine (NAC), an ROS scavenger. In addition, CSE reduced the migration ability of HBSMCs by 75%. The combination of NAC with CSE blocked the CSE-induced reduction of cell migration. However, CSE had no effect on TNF-${\alpha}$ secretion and NF-${\kappa}B$ activation. CSE induced an increase in intracellular $Ca^{2+}$ concentration in 64% of HBSMCs. CSE reduced the contractile ability of HBSMCs, and the ability was enhanced by NAC treatment. These results demonstrate that CSE treatment induces cell death and reduces migration and contraction by increasing ROS generation in normal HBSMCs. These results suggest that CSE may induce airway change through cell death and reduction in migration and contraction of normal HBSMCs.
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문제 정의
This study was performed to identify whether CSE changes airway remodeling-related processes, such as cell viability, migration, contraction, TNF-α release, or [Ca2＋]i in normal HBSMCs.
가설 설정
Arrowheads indicate the addition of CSE and/or NAC. (B) Effect of NAC on the CSE-induced Ca2＋ increase. The net change in Ca2＋ levels was normalized to F0 (Fmax-F0/F0), and the data obtained from all cells were averaged.
제안 방법
BDM was used as a contraction inhibitor (negative control). Images of the gels were captured with a camera (Canon, Tokyo, Japan), and the gel size change was measured at the indicated times with a ruler and quantified with image analysis software (Fluoview software program, Olympus). For each condition, collagen contraction was determined in at least quadruplicate.
(A) Migration ability of HBSMCs by CSE treatment. Phase-contrast images of wound areas at 0, 6, 12, and 24 h after 10% CSE treatment. (B) Representative microscopic images of migrating cells by 3-D migration assays.
changes, or cytokine release. To test whether CSE modulates the contractility of HBSMCs, a gel contraction assay was performed. The addition of 10% CSE to the HBSMC gels reduced contractility by 32% compared with the control (Fig.
데이터처리
A one-way ANOVA (SPSS18, Chicago, IL, USA) was used with p＜0.05 as the threshold for significance. Data are represented as mean±SD.
이론/모형
The cells’ migration ability was determined using a 3-D migration assay.
The cytotoxicity of CSE was examined in HBSMCs using an MTT assay. The HBSMCs were cultured in the presence of various concentrations of CSE (0.
성능/효과
In this study, CSE showed an inhibitory effect on cell proliferation, migration, and contraction of normal HBSMCs. However, we could not find a direct effect of CSE on TNF-α secretion or on the NF-κB activation of HBSMCs.
후속연구
Further studies will be needed to analyze the components of CSE extracted from different cigarettes to identify the molecular mechanism by which CSE affects cell proliferation, migration, TNF-α secretion, and contraction.
참고문헌 (45)
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