中和白细胞介素 17 对博来霉素诱导小鼠肺纤维化的干预作用

Protective effect of neutralizing interleukin-17 on Bleomycin- induced pulmonary fibrosis in mice

摘要:

目的??初步探讨中和白细胞介素 17（IL-17）对博来霉素（BLM）诱导小鼠肺纤维化形成过程的 干预作用及其机制。方法??96 只小鼠随机分为 4 组，即抗体干预组、同型抗体对照组（IgG2A 组） 、博来霉素 组（BLM 组） 、正常对照组（NC 组） 。气管内注射 BLM 使小鼠急性肺损伤诱导肺纤维化形成， NC 组则给予 等量生理盐水。复制模型后第 4、9、14、19 和 22 天抗体干预组小鼠尾静脉注射 IL-17A 中和抗体， IgG2A 组 注射同型抗体 IgG2A； BLM 组和 NC 组则注射等量生理盐水。在培养的第 7、 14 和 28 天将 4 组小鼠分批处死， 取气管肺泡灌洗液（BALF）涂片进行细胞计数， ELISA 法检测 IL-1β、 IL-10、 IL-17A、 IFN-γ 含量的变化； 检测小鼠肺组织羟脯氨酸（HYP） 、丙二醛（MDA）及活性氧（ROS）等生化指标，并评价各组小鼠肺组织 的纤维化程度。结果??抗体干预组与 BLM 组比较，BALF 中细胞总数、淋巴细胞、单核细胞、中性粒细胞的 个数， IL-1β、IL-10、IFN-γ 的含量， 以及 HYP、MDA、ROS 的含量在各个时间点均降低（ P <0.05） ； 而 IgG2A 组上述指标在各个时间点与 BLM 组比较，差异无统计学意义（ P >0.05） 。结论??中和 IL-17A 对 BLM 所致的实验性小鼠肺纤维化模型具有一定的防治作用， 有利于延缓肺纤维化的进程。

Abstract:

Objective To explore protective effect of neutralizing Interleukin-17 (IL-17) on Bleomycin (BLM)- induced pulmonary fibrosis in mice. Methods Totally 96 mice were randomly divided into antibody intervention group (antibody group), IgG group, BLM group and control group (n = 24). Endotracheal injection of BLM was performed resulting in acute lung injury and pulmonary fibrosis in mice. Mice in control group received normal saline. Mice were treated with IL-17 neutralizing antibody, IgG2A or BLM at day 4, 9, 14, 19, 22 in antibody group, IgG2A group and BLM group, respectively. Mice were sacrificed 7, 14, 21, 28 days, after cell culture, bronchoalveolar lavage fluid (BALF) was used to detect cell count. IL-1 β, IL-10, IL-17A and IFN-γ were measured by Elisa kits. Hydroxyprine (HYP), malonaldehyde (MDA) and reactive oxygen species (ROS) in lung tissue were determined. Histological characterization of pulmonary fibrosis was evaluated. Results Number of lymph, monocytes, neutrophils, concentration of IL-1β, IL-10, IFN-γ, IL-17A, and expression of HYP, MDA and ROS were decreased significantly in antibody group when compared with those in BLM group (P < 0.05 ). No obvious differences in the mentioned indexes were witnessed between IgG group and BLM group (P > 0.05). Conclusions Blockage of cytokine IL-17A effectively reduces BLM induced pulmonary fibrosis in mice through mediating inflammatory response.