转化生长因子β_1单克隆抗体干预成纤维细胞增殖的体外研究

The effect of anti transforming growth factor β 1 antibody on fibroblast proliferation in vitro

目的观察抗转化生长因子β１（ＴＧＦβ１）单克隆抗体对成纤维细胞（ＦＢ）增殖及胶原合成的影响。方法将肺泡巨噬细胞（ＡＭ）培养上清液与Ｌ９２９成纤维细胞株共同孵育，并加入不同剂量ＴＧＦβ１单克隆抗体，用３Ｈ胸腺嘧啶掺入法观察ＦＢ增殖及胶原合成的情况。结果抗ＴＧＦβ１单克隆抗体能够抑制ＡＭ条件上清液引起的ＦＢ增殖，在一定剂量范围内（１０μｇ／ｍｌ）呈剂量效应关系。抑制作用在１０μｇ／ｍｌ时达到最大并使ＦＢ培养上清Ⅳ型胶原的合成减少３２％。结论提示抗ＴＧＦβ１单克隆抗体的应用可能为肺纤维化的治疗提供潜在的新途径。

Objective Using anti transforming growth factor β 1 antibody binding to transforming growth factor β 1 (TGF β 1) in conditioned supernatant obtained from alveolar macrophages(AM) in broncho alveolar lavage fluid(BALF) in bleomycin induced rats,we investigated the effect of binding of TGF β 1 to anti TGF β 1 antibody on proliferation and synthesis of collagen by fibroblast in pulmonary fibrosis.Methods Bleomycin (0.75 mg/100 BW) was instilled intratracheally into wistar rats, then broncho alveolar lavage(BAL) was performed 7 days later. AMs (5×10 5/ml) were cultured in 0.2%FCS RPMI 1640 medium in vitro for 24 hours and conditioned medium was obtained. L929 fibroblast (7×10 3/well) was cultured in 96 well microtiter plate with conditioned medium and in different doses of anti TGF β 1 antibody(1、5、10、20 μg/ml) and IgG for 24 hours respectively. 3H Tdr was added 1 μCi per well for 6 hours before culture finished. The proliferation of fibroblast was studied by 3H Tdr incorporation rate and synthesis of collagen type Ⅳwas tested by ELISA. Results (1) Fibroblast proliferation which was induced by AM conditioned medium could be significantly depressed by anti TGF β 1 antibody in vitro( P < 0.01), and it appeared in a dose dependent manner. (2)Synthesis of collagen type Ⅳ was depressed about 32% by 10μg/ml anti TGF β 1 antibody.Conclusions Fibroblast proliferation and collagen synthesis could be depressed by anti TGF β 1 antibody in vitro. It seems possible to provide a new way for the treatment of pulmonary fibrosis.