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

ABSTRACT

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 (15) 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 (15) 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.