阻断白介素⁃17对博来霉素诱导小鼠肺纤维化及肺组织Bax/Bcl⁃2表达的影响
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张家口市科技局自然科学基金(12110046D?1);河北省高等学校科学研究重点项目(ZD2017206)


Influence of blocking of interleukin⁃17 on pulmonary fibrosis and expression of Bax/Bcl⁃2 in mice induced by bleomycin
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    摘要:

    目的:研究阻断白介素(interleukin,IL)-17对博来霉素诱导小鼠肺纤维化和肺组织Bax/Bcl-2表达的影响。方法:80只C57BL/6小鼠随机分为模型组、抗IL-17处理组、同型IgG处理组和PBS处理组。小鼠气管内一次性注入博来霉素诱导肺纤维化形成,PBS处理组给予等量生理盐水。4组小鼠分别从造模前1 d每隔3 d通过尾静脉给予抗鼠IL-17单克隆中和抗体或同型对照抗体或PBS。在造模后28 d,取各组小鼠肺组织,利用Masson染色及羟脯氨酸含量测定检测各组小鼠肺纤维化程度,通过流式细胞术测定肺组织细胞凋亡情况,采用免疫组织化学法检测肺组织Bax和Bcl-2的表达。结果:阻断IL-17后,小鼠肺纤维化程度明显降低(P < 0.01),羟脯氨酸含量显著下降(P < 0.01),细胞凋亡率明显降低(P < 0.01),Bax表达明显减弱(P < 0.01),Bcl-2表达虽无明显变化,但Bax/Bcl-2比值显著下降(P < 0.01)。结论:阻断内源性IL-17后,能显著降低博来霉素诱导的肺纤维化,显著降低肺组织细胞凋亡率和Bax/Bcl-2比值,这些数据提示阻断内源性IL-17活性改善博来霉素诱导的小鼠纤维化程度,可能与Bax/Bcl-2介导的线粒体细胞凋亡通路有关。

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    Objective:To study the influence on pulmonary fibrosis and expression of Bax/Bcl-2 in the lung after blocking interleukin(IL)-17 activity in mice induced by bleomycin(BLM). Methods:A total of 80 C57BL/6 mice were randomly divided into the following 4 groups,including the model group treated with neutralizing IL-17 antibody,the model group treated with isotype-matched control antibody,the model group and the PBS group. The three model groups were received a single intratracheal instillation of 5 mg/kg body weight of BLM in 0.05 mL sterile saline,while the PBS group were administrated the equivalent sterile saline. All mice were injected from the caudal vein with neutralizing rat antimouse IL-17 mAb,or control rat IgG,or PBS alone every 3 d starting on d1 before making models. At 28 d after model establishment,lung tissues from all mice were removed and used to measure the extent of pulmonary fibrosis by Masson staining and hydroxyproline contents measurement. The apoptosis rate of pulmonary cells was detected by flow cytometry(FCM). Meanwhile,the expressions of Bax/Bcl-2 in mice were also evaluated by immunohistochemistry(IHC). Results:Compared with other groups,the pulmonary fibrosis degree,hydroxyproline contents and apoptosis rate were significantly decreased in mice blocked with anti-IL-17 mAb(P < 0.01,respectively). Bax protein expression was decreased obviously(P < 0.01),although the Bcl-2 protein expression had no obvious changes,but the ratio of Bax/Bcl-2 decreased significantly(P < 0.01). Conclusion:Pulmonary fibrosis had been improved significantly after the endogenous IL-17 activity of mice blocked,the apoptosis rate and the ratio of Bax/Bcl-2 were all decreased in lung tissues. These data suggested that IL-17 improving the pulmonary fibrosis induced by BLM may be associated with Bax/Bcl-2 mediated mitochondrial apoptotic pathways.

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宋桂芹,徐志伟,王文栋,郝 敏,黄 勇,李向上,张效云.阻断白介素⁃17对博来霉素诱导小鼠肺纤维化及肺组织Bax/Bcl⁃2表达的影响[J].南京医科大学学报(自然科学版),2018,(3):317-321

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  • 收稿日期:2016-10-12
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  • 在线发布日期: 2018-03-19
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