摘要
目的了解矿物粉尘诱导的肺泡巨噬细胞（ＡＭ）因子在肺纤维化中的作用。方法用肺泡灌洗法获取兔ＡＭ，经体外培养，取其上清液，分别用同位素掺入法和四甲基偶氮唑盐比色法测定上清液中肿瘤坏死因子（ＴＮＦ）和白细胞介素６（ＩＬ６）的活性，并将此上清液与人胚肺成纤维细胞（ＷＩ３８）培养，用氚胸腺嘧啶（３ＨＴｄＲ）、１４碳脯氨酸（１４ＣＰｒｏ）测定成纤维细胞增殖和胶原合成，用氯氨Ｔ法测定总羟脯氨酸（ＨＯＰ）含量。结果石英、石棉和铀矿尘诱导的ＡＭ上清液均能促进肺成纤维细胞增殖和胶原合成，用３ＨＴｄＲ掺入的每分钟计数（ｃｐｍ）均值分别为６５８４、３８４８和６８９３（１００μｇ浓度组），１４ＣＰｒｏ掺入法ｃｐｍ均值分别为２７９５２、１３４１６和１８５３８（２００μｇ浓度组），明显高于对照组。ＡＭ上清液作用的肺成纤维细胞培养液总ＨＯＰ含量分别为２２．４１、２４．００和２１．３９μｇ／ｍｌ（２００μｇ浓度组），明显高于对照组（１２９１μｇ／ｍｌ）。石英、石棉和铀矿尘等矿物粉尘能刺激ＡＭ释放ＴＮＦ和ＩＬ６，其活性显著高于对照组，ＴＮＦ活性分别为１３９６、１１９８和８５２Ｕ／ｍｌ；ＩＬ６活性分别为１３３６、１５１?
Objective To understand the role of cytokine released from alveolar macrophage (AM) in lung fibrosis caused by mineral dust. Methods Rabbit′s AM obtained by lavage was cultured with mineral dust in vitro. Activities of tumor necrosis factor (TNF) and interleukin 6 (IL 6) in its supernatant were determined with isotope labelling method and MTT colorimetry, respectively. Human fetal lung fibroblast WI 138 was cultured with this supernatant. Proliferation of fibroblast and synthesis of collagen were examined by 3H thymidine ( 3H TdR) and 14 C proline ( 14 C Pro) incorporation and its total hydroxyproline (HOP) level was analyzed by chloramine T method. Results Proliferation of lung fibroblast and synthesis of collagen could be enhanced by the supernatant containing AM induced by quartz, asbestos and uranium mineral dust, with 3H TdR incorporated counts per minute (cpm) of 6 584, 3 848 and 6 893 in the group of 100 μg 3H TdR and 14 C Pro incorporated 27 952, 13 416 and 18 538 in the group of 200 μg respectively, which were significantly higher than those in the control group. Total HOP levels in the culture media for lung fibroblast enhanced by AM supernatant were 22.41, 24.00 and 21.39 μg/ml, respectively, and was significantly higher than that in the control group (12.91 μg/ml). Release of TNF and IL 6 could be stimulated by mineral dust, such as quartz, asbestos and uranium mineral dust, and their activities were significantly higher than those in the control group, with those of 1 396, 1 198 and 852 U/ml in TNF group and 1 336, 1511 and 1 335 U/ml in IL 6 group, respectively. Proliferation of lung fibroblast and synthesis of collagen could be inhibited by antibody against TNF and interferon r (gamma), and the effect of the latter was weaker than that of the former on inhibition of fibroblast proliferation and the effect on collagen synthesis was just in the opposite direction. Conclusion Lung fibrosis caused by mineral dust correlated with abnormal expression of TNF and IL 6. Antibody against TNF and gamma interferon could antagonize the effect of NTF and IL 6.
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
1998年第6期336-339,共4页
Chinese Journal of Preventive Medicine
