Tuberc Respir Dis.  2009 Dec;67(6):528-535.

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

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

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 micrometer (PM(10)) is associated with the exacerbation of respiratory diseases and increased mortality of heart and lung disease patients. The fibrotic effects of PM(10) of Asian dust to pulmonary fibroblast cells are unknown. This study examined the production of reactive oxygen species (ROS), TGF-beta, NF-kappaB, PDGF-alpha 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 microgram/mL of PM(10) for 24 hours. ROS was detected by measuring the level of oxidized DCF using FACS. TGF-beta, NF-kappaB, PDGF-alpha and fibronectin were detected by western blotting. RESULTS: There was no increase in the ROS, TGF-beta and PDGF-alpha levels in the MRC-5 cells exposed to PM(10). The NF-kappaB level was higher in the MRC-5 cells exposed to 50 and 100 microgram/mL of PM(10) for 24 hours. The fibronectin level in the MRC-5 cells after 24 hours incubation with 50 microgram/mL PM(10) was significantly higher than the control group (PM(10) 50 microgram/mL 113.27+/-8.65 of control, p=0.005). CONCLUSION: PM(10) from Asian dust increases the activation of NF-kappaB and fibronectin expression in MRC-5 fibroblast cells.

Keyword

Particulate Matter; Dust; Reactive Oxygen Species; Pulmonary Fibrosis; Fibroblasts

MeSH Terms

Asian Continental Ancestry Group
Blotting, Western
China
Dust
Fibroblasts
Fibronectins
Heart
Humans
Lung Diseases
Mongolia
NF-kappa B
Particulate Matter
Pulmonary Fibrosis
Reactive Oxygen Species
Transforming Growth Factor beta
Wind
Dust
Fibronectins
NF-kappa B
Particulate Matter
Reactive Oxygen Species
Transforming Growth Factor beta

Figure

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  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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