[Effect of herbs 814 on tumor necrosis factor a production from alveolar macrophages in vitro].

OBJECTIVE

To study the influence of herbs 814 on the secretion of tumor necrosis factor alpha (TNF-alpha) from hamster alveolar macrophages (AMs) in vitro and elucidate mechanism of herbs 814's prevention of emphysema.

METHODS

AMs of hamster were collected by bronchial alveolar lavage (BAL). The lung lavage cells were adjusted to 5 x 10(5)/ml and seeded in each well of a 24-well tissue culture plates. The herbs 814 was added into these wells before or after lipopolysaccharide (LPS) stimulation and then cultured supernatants were collected. TNF-alpha production in the supernatants was tested by ELISA, TNF-alpha cytotoxicity was assayed using L929 cells which were susceptible to TNF-alpha and a monoclonal antibody (MAb) that neutralizes rhTNF-alpha was utilized to identify the cytotoxicity of cultured TNF-alpha.

RESULTS

ELISA showed that TNF-alpha production in the supernatants with 814 added was lower than those either in LPS-stimulated or non LPS-stimulated supernatants and there was a significant decrease in the supernatants of higher concentration (1:5) of herbs 814. Cytotoxicity test showed that TNF-alpha cytotoxic activity in the supernatants into which herbs 814 was added was lower than one in the LPS-stimulated supernatants and there was a significant decrease in the supernatants of higher concentration (1:5) of herbs 814. Meanwhile, MAb significantly reduced cytotoxicity of LPS-stimulated culture supernatants.

CONCLUSIONS

Herbs 814 could inhibit the secretion of TNF-alpha and prevent the lung damage mediated by TNF-alpha.