IL-17A抗体对成纤维细胞增殖及分泌羟脯氨酸的影响
The effect of IL-17A antibody on proliferation and hydroxyproline secretion of fibroblasts
张明 1黄成亮 1王文军 1马艳梅 1范贤明1
作者信息
- 1. 646000,泸州医学院附属医院呼吸内科
- 折叠
摘要
目的 观察IL-17A抗体对成纤维细胞增殖及分泌羟脯氨酸的影响.方法 将清洁级雌性Wistar大鼠分为生理盐水(normal saline,NS)组和博来霉素(bleomycin,BLM)组.NS组、BLM组大鼠分别于气管内灌注NS和BLM,并于气管内灌注后第7天心脏采血处死,右肺行灌洗,并分离、纯化肺泡巨噬细胞(alveolar macrophage,AM),将其培养24 h后分别取2组AM培养上清;左肺行病理切片及HE染色.将大鼠成纤维细胞(208F细胞)分为对照组、模型组、抗体组(A组、B组、C组)培养,各组培养基分别为:NS组AM培养上清、BLM组AM培养上清、BLM组AM培养上清+不同质量浓度IL-17A抗体(0.5、1、2μg/L),分别于第24、48和72小时用CCK-8检测208F细胞增殖.208F细胞按上述方法分为5组培养24 h后,应用酶联免疫分析(ELISA)检测208F培养上清中的羟脯氨酸含量.结果 BLM组大鼠肺部炎症明显高于NS组.208F细胞增殖结果显示:模型组和抗体组各组在各个时间点均高于对照组(P＜0.05);各个抗体组在各个时间点均低于模型组(P＜0.05);抗体A组在各个时间点均高于其他两抗体组(P＜0.05),抗体B组和抗体C组在各个时间点均无明显差异(P＞0.05).208F细胞培养上清中羟脯氨酸结果显示:模型组和各个抗体组均高于对照组(P＜0.05);各个抗体组均低于模型组(P＜ 0.05);抗体A组高于其他两抗体组(P＜0.05),抗体B组和抗体C组无明显差异(P＞0.05).结论 在体外IL-17A抗体可减轻肺纤维化大鼠AM诱导的成纤维细胞过度增殖和胶原蛋白过度表达.
Abstract
To investigate the effect of IL-17A antibody on the proliferation and hydroxyproline secretion of fibroblasts,healthy female Wistar rats were randomly divided into normal saline (NS) group and bleomycin (BLM) group.Intratracheal instillation of BLM was given to BLM group and intratracheal instillation of physiological saline was given to NS group.The rats of two groups were sacrificed respectively at day 7 after the intratracheal instillation by exsanguinations of right ventricle of heart under general anesthesia,and then bronchoalveolar lavage (BLF) of the right lung was performed to obtain alveolar macrophages (AMs),while the left lung was used for pathological HE staining.Cultured medium was collected when the AMs cultured for 24 hours.Then rats' fibroblasts (208F) were divided into 5 groups:controlled group (cultured medium of NS group's AMs) and model group (cultured medium of BLM group AMs),the antibody group (groups A,B and C:cultured medium of BLM group AMs + IL-17A antibody of different concentrations of 0.5,1 and 2 g/L,respectively).Hydroxyproline in the cultured medium of 208F was test by ELISA after 24 hours culturation; the proliferation of 208F cells was respectively detected at 24 hours,48 hours and 72 hours after cultivation by CCK-8.We found that the lung inflammation of rats of BLM group was higher than that in group NS.As to 208F proliferation:the model group and the antibody group were higher than the control group at each time point (P＜0.05); each antibody group was lower than model group at different time points (P＜ 0.05); group A were higher than that of the other two antibody groups at each time point (P＜ 0.05),group B and group C were not significantly different at each time point (P＞ 0.05).As to the hydroxyproline level in the 208F cell culture supernatant:model group and each antibody group were higher than control group (P ＜ 0.05); each antibody group was lower than the model group (P＜ 0.05);group A were higher than the other two antibody groups (P＜ 0.05),groups B and C had no significant difference (P＞ 0.05).All the results show that IL-17A antibody can reduce the excessive proliferation and collagen overexpression of fibroblasts induced by alveolar macrophages of pulmonary fibrosis of rat in vitro.
关键词
IL-17A抗体/肺泡巨噬细胞/成纤维细胞/细胞增殖/羟脯氨酸Key words
IL-17A antibody/Alveolar macrophages/Fibroblasts/Proliferation/Hydroxyproline引用本文复制引用
基金项目
泸州市科技局资助项目(12102)
泸州医学院附属医院青年基金(14053)
出版年
2014
NETL
NSTL
维普
万方数据