中药814对肺泡巨噬细胞分泌肿瘤坏死因子的影响

Effect of Herbs 814 on Tumor Necrosis Factor a Production from Alveolar Macrophages in vitro

目的 研究中药８１４对肺泡巨噬细胞（ＡＭｓ）分泌肿瘤坏死因子（ＴＮＦ－α）的影响进而探讨８１４对肺气肿的作用机理。方法 支气管肺泡灌洗（ＢＡＬ）收集金黄地鼠ＡＭｓ，调整细胞浓度为５×１０＾５／ｍｌ，接种于２４孔板培养，在有多糖（ＬＰＳ）刺激前后加入中药８１４分别收集距养细胞上清液，通过ＥＬＩＳＡ测定上清液ＴＮＦ－α含量，以及运用ＴＮＦ－α敏感的小鼠Ｌ９２９细胞株。

Objective To study the influence of herbs 814 on the secretion of tumor necrosis factor α (TNF-α) 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 5×10~5/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 cultureed supernatants were collected. TNF-α production in the supernatants was tested by ELISA, TNF-α cytotoxicity was assayed using L929 ceils which were susceptible to TNF-α and a monoclonal antibody (MAb) that neutralizes rhTNF-α was utilized to identify the cytotoxicity of cultureed TNF-α. Results ELISA showed that TNF-α 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-α 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-α and prevent the lung damage mediated by TNF-α.