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Li, Lee, Park, Chang, Lee, Kim, Lee, and Choe: The Effects of Air-borne Particulate Matters on the Alveolar Macrophages for the TNF-α and IL-1β Secretion
Original Article
Tuberculosis and Respiratory Diseases

Tuberculosis and Respiratory Diseases 2006; 60(5): 554-563.

Published online: 31 May 2006

DOI: https://doi.org/10.4046/trd.2006.60.5.554

The Effects of Air-borne Particulate Matters on the Alveolar Macrophages for the TNF-α and IL-1β Secretion

Tian Zhu Li1, Soo-Jin Lee1, Se-Jong Park1, Byung-Joon Chang1, Jong-Hwan Lee1, Kil-Soo Kim2, Myoung-Heon Lee3, Nong-Hoon Choe1

1Konkuk University College of Veterinary Medicine, Seoul, Korea.

2Kyungpook National University College of Veterinary Medicine, Korea.

3National Veterinary Research and Quarantine Service, Korea.

Address for correspondence: Nong-Hoon Choe, PhD. Konkuk University College of Veterinary Medicine 1 Hwa-yang dong, Gwang-jin gu, Seoul, Korea. Phone: 02-450-3709, Fax: 02-450-3037, nojamaji@hanmail.net

Received 13 December 2005       Accepted 2 May 2006

Copyright © 2006 The Korean Academy of Tuberculosis and Respiratory Diseases

Abstract

Background

PM is known to induce various pulmonary diseases, including asthma, cancer, fibrosis and chronic bronchitis. Despite the epidemiological evidence the pathogenesis of PM-related pulmonary diseases is unclear.

Methods

This study examined the effects of PM exposure on the secretion of TNF-α and IL-1β in the cultured alveolar macrophages. The cultured primary alveolar macrophages were treated with the medium, PM (5~20µg/cm2), LPS (5ng/ml), and PM with LPS for 24h and 48h respectively. ELISA was used to assay the secreted TNF-α and IL-β in the culture medium. Western blotting was used to identify and determine the level of proteins isolated from the culture cells. The cells cultured in the Lab-Tek® chamber slides were stained with immunocytochemical stains.

Results

PM induced TNF-α and IL-1β secretion in the culturing alveolar macrophages, collected from the SPF and inflammatory rats. However, the effects were only dose-dependent in the inflammatory macrophages. When the cells were co-treated with PM and LPS, there was a significant synergistic effect compared with the LPS in the both cell types.

Conclusion

PM might be play an important role in the induction and/or potentiation of various lung diseases by oversecretion of TNF-α and IL-1β.

Keywords: Particulate matter (PM), Alveolar macrophages, Tumor necrosis factor-α (TNF-α), Interleukin 1β (IL-1β)

Figures and Tables

Figure 1
Treatment-response relation for TNF-α secretion in the cultured inflammatory BAL cells. Cells were cultured with medium only, LPS (10ng/ml) only and various concentrations of PM (5~20µg/cm2) for 24 hours and 48 hours.
**P<0.01 versus control. Mean ± SEM of four independent experiments per category.
trd-60-554-g001
Figure 2
Treatment-response relation for IL-1β secretion in the cultured inflammatory BAL cells. Cells were cultured with medium only, LPS (10ng/ml) only and various concentrations of PM (5~20µg/cm2) for 24 hours and 48 hours.
**P<0.01 versus control. Mean ± SEM of four independent experiments per category.
trd-60-554-g002
Figure 3
Treatment-response relation for TNF-α secretion in the cultured SPF rat alveolar macrophages at 24 hours and 48 hours. Cells were cultured with medium only (sham control), LPS (5ng/ml), various concentrations of PM (5, 10, 20µg/cm2) and various concentrations of PM with LPS.
*P<0.05, **P<0.01 versus LPS; †P<0.05 versus control. Mean ± SEM of four independent experiments per category (each experiment's n=3)
trd-60-554-g003
Figure 4
Treatment-response relation for IL-1β secretion in the cultured SPF rat's alveolar macrophages at 24 hours and 48 hours. Cells were cultured with medium only (sham control), LPS (5ng/ml), various concentrations of PM (5, 10, 20µg/cm2) and various concentrations of PM with LPS.
*P<0.05, **P<0.01 versus LPS; †P<0.05 versus control. Mean ± SEM of four independent experiments per category (each experiment's n=3)
trd-60-554-g004
Figure 5
Immunocytochemical stains for TNF-α in the cultured rat alveolar macrophages (× 400). Cells were cultured with (a) medium only, (b) LPS (5ng/ml) only, (c) PM (20µg/cm2) only, and (d) LPS with PM at 6h. (e) LPS only (× 1,000) and (f) LPS with PM (× 1,000) at same incubation time.
trd-60-554-g005
Figure 6
Immunocytochemical stains for IL-1β in the cultured rat alveolar macrophages (× 400). Cells were cultured with (a) medium only, (b) LPS (5ng/ml) only, (c) PM (20µg/cm2) only, and (d) LPS with PM at 6h. (e) LPS only (× 1,000) and (f) LPS with PM(× 1,000) at same incubation time.
trd-60-554-g006
Figure 7
Western blot for TNF-α and IL-1β in the cultured rat alveolar macrophages. Cells were cultured with medium only, LPS only, PM only and PM with LPS for 6 hours. The level of b-actin was similar in all of the tested samples.
trd-60-554-g007
Table 1
Conditions of microwave digestion system
trd-60-554-i001
Table 2
Analysis conditions of PM
trd-60-554-i002
Table 3
PM analysis
trd-60-554-i003
Table 4
Proportion(%) of positive cells to TNF-α and IL-1β in cultured alveolar macrophages after treatment with medium only, LPS (5ng/ml) only, PM (20µg/cm2) only and PM with LPS at 6 hours.
trd-60-554-i004

**P<0.01 versus control.

†P<0.01 versus LPS. (each experiment's n=3, Mean ± SEM).

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