Tuberc Respir Dis.  2012 Aug;73(2):84-92.

Asian Dust Particles Induce TGF-beta1 via Reactive Oxygen Species in Bronchial Epithelial Cells

• Kyung SY
• Yoon JY
• Kim YJ
• Lee SP
• Park JW
• Jeong SH

Abstract

BACKGROUND
Asian dust storms can be transported across eastern Asia. In vitro, Asian dust particle-induced inflammation and enhancement of the allergic reaction have been observed. However, the fibrotic effects of Asian dust particles are not clear. Production of transforming growth factor beta1 (TGF-beta1) and fibronectin were investigated in the bronchial epithelial cells after exposure to Asian dust particulate matter (AD-PM10).
METHODS
During Asian dust storm periods, air samples were collected. The bronchial epithelial cells were exposed to AD-PM10 with and without the antioxidant, N-acetyl-L-cysteine (NAC). Then TGF-beta1 and fibronectin were detected by Western blotting. The reactive oxygen species (ROS) was detected by the measurement of dicholorodihydrofluorescin (DCF), using a FACScan, and visualized by a confocal microscopy.
RESULTS
The expression of TGF-beta1, fibronectin and ROS was high after being exposed to AD-PM10, compared to the control. NAC attenuated both TGF-beta1 and fibronectin expression in the AD-PM10-exposed the bronchial epithelial cells.
CONCLUSION
AD-PM10 may have fibrotic potential in the bronchial epithelial cells and the possible mechanism is AD-PM10-induced intracellular ROS.

Keyword

Air Pollutants; Reactive Oxygen Species; Transforming Growth Factor beta

MeSH Terms

Acetylcysteine
Air Pollutants
Asian Continental Ancestry Group
Blotting, Western
Dust
Epithelial Cells
Far East
Fibronectins
Humans
Hypersensitivity
Inflammation
Particulate Matter
Reactive Oxygen Species
Transforming Growth Factor beta
Transforming Growth Factor beta1
Acetylcysteine
Air Pollutants
Dust
Fibronectins
Particulate Matter
Reactive Oxygen Species
Transforming Growth Factor beta
Transforming Growth Factor beta1

Figure

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  Figure 1 The proportion of chemical components in AD-PM10 used in this study. This was analyzed at the Korea Institute of Ceramic Engineering and Technology.

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  Figure 2 Time-dependent and dose-dependent effects of AD-PM10 on transforming growth factor β1 (TGF-β1) and fibronectin production. WI-26VA4 cells were incubated for the indicated period of time with 100 µg/mL of AD-PM10 (A). Alternatively, these cells were incubated for 48 hours with the indicated concentrations of AD-PM10 (B). The levels of TGF-β1 production with 50 and 100 µg/mL of AD-PM10 were significantly higher compared with control values (black squares). The levels of fibronectin production with AD-PM10 were significantly higher compared with the control values (white circle). In the dose-response experiments of AD-PM10, the means±SD of five separate experiments are shown. *Significant increase over the non-stimulated control cells.

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  Figure 3 Dicholorodihydrofluorescin (DCF)-sensitive reactive oxygen species (ROS) production in bronchial epithelial cells incubated with 0~500 µg/mL of a AD-PM10 suspension. The confocal microscopic examination shows a high expression of intracellular ROS in AD-PM10-exposed bronchial epithelial cells (A). On measuring the levels of DCF-sensitive ROS using FACScan, ROS in AD-PM10-exposed bronchial epithelial cells showed significantly higher expression compared with controls (black squares) and N-acetyl-L-cysteine (NAC) attenuated the increase of ROS generation (white squares) (B). The experiments were repeated over five times. *Significant increase over unstimulated control cells. †Significant differences between NAC-treated cells and cells not treated with NAC.

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  Figure 4 Effect of NAC on AD-PM10-induced transforming growth factor β1 (TGF-β1). On Western blotting, TGF-β1 with exposure to AD-PM10 increased more than the unexposed control, and N-acetyl-L-cysteine (NAC) attenuated TGF-β1 induction with AD-PM10 (A). The levels of TGF-β1 production with 50 and 100 µg/mL of AD-PM10 were significantly higher compared with control values (white bar). NAC effectively blocked the expression of TGF-β1 (black bar) (B). The experiments were repeated over five times. *A significant increase over unstimulated control cells. †,‡Significant difference between cells treated with NAC and cells not treated with NAC (†p<0.05, ‡0.05

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  Figure 5 Effect of N-acetyl-L-cysteine (NAC) on AD-PM10 induced fibronectin. On Western blotting, fibronectin with exposure to AD-PM10 increased greater than the unexposed control, and NAC attenuated fibronctin production with AD-PM10 (A). The levels of fibronectin production with 50 and 100 µg/mL of AD-PM10 were significantly higher compared with control values (white bar). And NAC effectively blocked the expression of fibronectin (black bar) (B). The experiments were repeated over five times.

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