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Kim, Chon, Yoon, Kim, Kyung, Lee, Park, and Jeong: The Effect of Particulate Matter 10 from Asian Dust on the Production of Reactive Oxygen Species, TGF-β, NF-κB, PDGF-α and Fibronectin in MRC-5 Fibroblast Cells
Original Article
Tuberculosis and Respiratory Diseases

Tuberculosis and Respiratory Diseases 2009; 67(6): 528-535.

Published online: 31 December 2009

DOI: https://doi.org/10.4046/trd.2009.67.6.528

The Effect of Particulate Matter 10 from Asian Dust on the Production of Reactive Oxygen Species, TGF-β, NF-κB, PDGF-α and Fibronectin in MRC-5 Fibroblast Cells

Ah Hyun Kim, M.D.1, Suyeon Chon, M.D.2, Jin Young Yoon2, Yu Jin Kim, M.D.2, Sun Young Kyung, M.D.2, Sang Pyo Lee, M.D.2, Jeong Woong Park, M.D.2, Sung Hwan Jeong, M.D., Ph.D.2

1Gachon University of Medicine and Science, Incheon, Korea.

2Division of Pulmonology, Department of Internal Medicine, Gachon University Gil Hospital, Incheon, Korea.

Address for correspondence: Sung Hwan Jeong, M.D. Division of Pulmonology, Department of Internal Medicine, Gachon University Gil Medical Center, 1198, Kuwol-dong, Namdong-gu, Incheon 405-760, Korea. Phone: 82-32-460-3201, Fax: 82-32-469-4320 jsw@gilhospital.com

Received 14 September 2009       Accepted 4 November 2009

Copyright©2009. The Korean Academy of Tuberculosis and Respiratory Diseases. All rights reserved.

Abstract

Background

Dust clouds blown by the wind from the arid deserts of Mongolia and Northeast China are known as Asian dust storms. Ambient particulate matter with a diameter <10 µm (PM10) is associated with the exacerbation of respiratory diseases and increased mortality of heart and lung disease patients. The fibrotic effects of PM10 of Asian dust to pulmonary fibroblast cells are unknown. This study examined the production of reactive oxygen species (ROS), TGF-β, NF-κB, PDGF-α and Fibronectin in fibroblasts exposed to Asian dust particles.

Methods

Air samples were collected using a high volume air sampler (Sibata model HV500F) with an air flow of 500 L/min for at least 6 hours. The MRC-5 cells were exposed to 0, 50 and 100 µg/mL of PM10 for 24 hours. ROS was detected by measuring the level of oxidized DCF using FACS. TGF-β, NF-κB, PDGF-α and fibronectin were detected by western blotting.

Results

There was no increase in the ROS, TGF-β and PDGF-α levels in the MRC-5 cells exposed to PM10. The NF-κB level was higher in the MRC-5 cells exposed to 50 and 100 µg/mL of PM10 for 24 hours. The fibronectin level in the MRC-5 cells after 24 hours incubation with 50 µg/mL PM10 was significantly higher than the control group (PM10 50 µg/mL 113.27±8.65 of control, p=0.005).

Conclusion

PM10 from Asian dust increases the activation of NF-κB and fibronectin expression in MRC-5 fibroblast cells.

Keywords: Particulate Matter, Dust, Reactive Oxygen Species, Pulmonary Fibrosis, Fibroblasts

Figures and Tables

Figure 1
Measured Reactive oxygen species (ROS) by FACS in MRC-5 cells after 24 hours incubation with PM10. MRC-5 cells were exposed to 50 and 100 µg/mL of ambient particulate matter with a diameter of less than 10 µm for 24 hours. Results shown in the box graph are measured ROS values by FACS. The measured ROS in MRC-5 cells after 24 hours incubation with PM10 were not increased (PM10 0 µg/mL 113.01±83.4, PM10 50 µg/mL 92.48±70.46, p=0.674, PM10 100 µg/mL 106.80±88.73, p=0.916).
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Figure 2
Confocal microscopic findings in MRC-5 cells after 24 hours incubation with PM10. The fluorogenic probe 2'7'-dichlordihydrofluorescin diacetate (DCF-DA) in the cell was oxidized and detected as green color. The oxidized DCF-DA was not increased in MRC-5 cells after 24 hours incubation with PM10.
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Figure 3
TGF-β in MRC-5 cells after 24 hours incubation with PM10. TGF-β was detected by western blotting. The density of band was measured by densitometry. Results shown in the bar graph are the percentage from control values. TGF-β in MRC-5 cells after 24 hours incubation with PM10 was not increased compared with control group (PM10 50 µg/mL 101.43±6.99 of control, p=1.000, PM10 100 µg/mL 97.91±26.40 of control, p=1.000).
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Figure 4
NF-κB in MRC-5 cells after 24 hours incubation with PM10. MRC-5 cells were exposed to 50 and 100 µg/mL of ambient particulate matter with a diameter of less than 10 µm for 24 hours. NF-κB was detected by western blotting.
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Figure 5
PDGF-α in MRC-5 cells after 24 hours incubation with PM10. PDGF-α was detected by western blotting. The density of band was measured by densitometry. Results shown in the bar graph are the percentage from control values. Results shown in the bar graph are the percentage from control values (*p<0.05). PDGF-α in MRC-5 cells after 24 hours incubation with 50 µg/mL PM10 was not significantly increased compared with control group (PM10 50 µg/mL 101.94±8.13 of control, p=0.487). PDGF-α in MRC-5 cells with 100 µg/mL PM10 was signifi cantly decreased compared with control group (PM10 100 µg/mL 97.50±1.81 of control, p=0.037).
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Figure 6
Fibronectin and β-actin in MRC-5 cells after 24 hours incubation with PM10. MRC-5 cells were exposed to 50 and 100 µg/mL of ambient particulate matter with a diameter of less than 10 µm for 24 hours. Fibronectin was detected by western blotting. The density of band was measured by densitometry. Results shown in the bar graph are the percentage from control values (*p<0.05). Fibronectin in MRC-5 cells after 24 hours incubation with 50 µg/mL PM10 was significantly increased compared with control group (PM10 50 µg/mL 113.27±8.65 of control, p=0.005). Fibronectin in MRC-5 cells with 100 µg/mL PM10 was not increased compared with control group (PM10 100 µg/mL 107.82±11.91 of control, p=0.577).
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Table 1
Analysis of Asian sand dust
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Korea Institute of Ceramic Engineering and Technology (KICET).

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