羧胺三唑对博莱霉素致小鼠肺纤维化的治疗作用及其机制

Therapeutic effect and mechanism of carboxyamidotriazole on the pulmonary fibrosis of mice induced by bleomycin

目的探讨羧胺三唑（CAI）对博莱霉素致小鼠肺纤维化的治疗作用，并探讨其可能的机制。方法将45只小鼠按随机数字表法随机分为3组：（1）空白组：一次性尾静脉内注射生理盐水0.2 ml后，按0.1 ml/10 g给予聚乙二醇400溶液（PEG－400）灌胃，1次/d，连续14 d；（2）博莱霉素组：一次性尾静脉内注射博莱霉素150 mg/kg，后按0.1 ml/10 g给予PEG－400溶液灌胃，1次/d，连续14 d；（3）羧胺三唑组：一次性尾静脉内注射博莱霉素150 mg/kg，后给予羧胺三唑溶液40 mg/kg灌胃，1次/d，连续14 d。观察28 d，内容包括：肺系数、生存分析、肺组织病理切片和胶原染色、肺组织羟脯氨酸含量以及肺组织匀浆中转化生长因子β1（TGF－β1）、γ干扰素（IFN－γ）、基质金属蛋白酶9（MMP－9）和基质金属蛋白酶抑制剂1（TIMP－1）含量。结果第28天时博莱霉素组和羧胺三唑组肺系数显著高于空白组，以博莱霉素组最高（均P〈0.05）。空白组、羧胺三唑组、博莱霉素组肺组织纤维化程度依次加重；空白组、博莱霉素组、羧胺三唑组肺组织羟脯氨酸含量依次为（0.406±0.020）、（0.722±0.118）、（0.537±0.071）μg/mg（均P〈0.05）；3组肺组织匀浆中TGF－β1含量依次为（15±5）、（60±10）、（41±10）ng/ml（均P〈0.05），IFN－γ含量依次为（47±5）、（126±24）、（194±34）pg/ml（均P〈0.05），TIMP－1含量依次为（73±6）、（369±58）、（246±51）ng/ml（均P〈0.05）；而MMP－9含量差异无统计学意义（P〉0.05）。结论羧胺三唑可减轻博莱霉素致小鼠肺纤维化，其机制可能与TGF－β1、IFN－γ、MMP－9和TIMP－1等细胞因子有关。

ObjectiveTo investigate the treatment effect of carboxyamidotriazole （CAI） on bleomycin induced lung fibrosis in mice, and the potential mechanism involved.MethodsA total of 45 mice were divided into three groups randomly. Blank control group （blank group）： after a one-time tail vein injection of saline solution 0.2 ml, mice were given polyethylene glycol 400 （PEG-400） 0.1 ml/10 g by gavage once daily for 14 days; the bleomycin group （BLM group）： after a one-time tail vein injection of bleomycin 150 mg/kg, mice were given PEG-400 solution 0.1 ml/10 g by gavage once daily for 14 days; CAI group： after a one-time tail vein injection of bleomycin 150 mg/kg, mice were given CAI solution 40 mg/kg by gavage once daily for 14 days. All mice were sacrificed on day 28. Observation index： lung coefficient, survival analysis, pathological section and collagen staining of lung tissue, lung hydroxyproline, Transformation growth factor-β1（TGF-β1）, γ-interferon（IFN-γ）, matrix metalloproteinase 9（MMP-9） and tissue inhibitor of matrix metalloproteinasese 1（TIMP-1） content determination in lung homogenate.ResultsOn day 28 the lung coefficient of mice in BLM group and CAI group was significantly higher than the blank group, and the BLM group was with the highest （all P〈0.05）. Degree of pulmonary fibrosis in lung tissue pathological specimens （HE staining） was, from heavy to light, BLM group, CAI group, blank group. The content of hydroxyproline in mice lung homogenate was （0.406±0.020） μg/mg in blank group, （0.722±0.118） μg/mg in BLM group, （0.537±0.071） μg/mg CAI group, respectively （all P〈0.05）. The content of TGF-β1 in three groups was （15±5）, （60±10）, （41±10） ng/ml respectively （all P〈0.05）. The content of IFN-γ in three groups was （47±5）, （126±24）, （194±34） pg/ml respectively （all P〈0.05）. The content of TIMP-1 in three groups was （73±6）, （369±58）, （246±51） ng/ml respectively （all P〈0.05）. Comparisons of the content of MMP-9 between each group had no significant difference （P〉0.05）.ConclusionsCAI can reduce lung injury induced by bleomycin in mice. The mechanism of action is related to the effects of CAI on cytokines such as TGF-β1, IFN-γ, MMP-9 and TIMP-1.