Tuberc Respir Dis.  2006 May;60(5):554-563.

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

• Li TZ
• Lee SJ
• Park SJ
• Chang BJ
• Lee JH
• Kim KS
• Lee MH
• Choe NH

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-alpha and IL-1beta in the cultured alveolar macrophages. The cultured primary alveolar macrophages were treated with the medium, PM (5~20 microgram/cm2), LPS (5ng/ml), and PM with LPS for 24h and 48h respectively. ELISA was used to assay the secreted TNF-alpha and IL-beta 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(R) chamber slides were stained with immunocytochemical stains.
RESULTS
PM induced TNF-alpha and IL-1beta 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-alpha and IL-1beta.

Keyword

Particulate matter (PM); Alveolar macrophages; Tumor necrosis factor-alpha (TNF-alpha); Interleukin 1beta (IL-1beta)

MeSH Terms

Animals
Asthma
Blotting, Western
Bronchitis, Chronic
Coloring Agents
Enzyme-Linked Immunosorbent Assay
Fibrosis
Lung Diseases
Macrophages
Macrophages, Alveolar*
Rats
Tumor Necrosis Factor-alpha*
Coloring Agents
Tumor Necrosis Factor-alpha

Figure

• 

  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.

• 

  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.

• 

  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)

• 

  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)

• 

  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.

• 

  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.

• 

  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.

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