阻断白介素-17对博来霉素诱导小鼠肺纤维化及肺组织Fas/FasL表达的影响
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张家口市科技局自然科学基金资助(12110046D-1) ;河北省高等学校自然科学重点项目(ZD2017206)


Blocking effects of interleukin-17 on pulmonary fibrosis and expression of Fas/FasL in mice induced by bleomycin
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    摘要:

    目的:探讨阻断白介素-17(interleukin-17,IL-17)对博来霉素(bleomycin,BLM)诱导小鼠肺纤维化、细胞凋亡和肺组织Fas/FasL表达的影响。方法:80只C57BL/6小鼠随机分为4组,分别为假手术组(SG)、BLM组(BG)、中和抗体组(NG)和抗体对照组(IG),经小鼠气管内一次性注入BLM诱导肺纤维化的形成,假手术组则给予等量生理盐水。NG和IG小鼠分别从造模前1天每隔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,SG和BG则给予等量PBS。于造模后28 d,处死各组小鼠,取肺组织,通过Masson染色及羟脯氨酸含量测定评价各组小鼠肺纤维化程度,用流式细胞仪和免疫组织化学法分别检测肺组织细胞凋亡和Fas/FasL的表达情况。结果:与BG和IG相比,NG小鼠肺纤维化程度明显减弱(P<0.01),羟脯氨酸含量显著下降(P<0.01),细胞凋亡率和Fas/FasL表达亦显著降低(P<0.01)。结论:内源性IL-17被阻断后,能显著改善BLM诱导的肺纤维化,降低细胞凋亡率和Fas/FasL表达,表明阻断IL-17对BLM诱导的小鼠纤维化改善与Fas/FasL凋亡通路有关。

    Abstract:

    Objective:To explore the effect on pulmonary fibrosis,cell apoptosis and expression of Fas/FasL in lung by blocking IL-17 activity in mice induced by bleomycin(BLM). Methods:Eighty C57BL/6 mice were randomly divided into the following 4 groups:sham group(SG),BLM group(BG),neutralizing antibody group(NG) and isotype-matched control antibody group(IG),respectively. Three groups were received a single intratracheal instillation of 5 mg/(kg body weight) of BLM to induce pulmonary fibrosis,while SGs was administrated the equvalent sterile saline. NG and IG were injected through caudal vein with neutralizing rat antimouse IL-17 mAb,or control rat IgG every 3 days starting on day 1 before making model,SG and BG were received equvalent PBS alone. All mice were sacrificed after 28 days. Lung tissues were removed and used to evaluate the extent of pulmonary fibrosis by Masson staining and hydroxyproline contents measurement. Cell apoptotic rate and the expression of Fas/FasL in mice were detected through Flow cytometry and immunohistochemical method. Results:Compared with BG and IG,the pulmonary fibrosis degree of NG was decreased remarkably(P<0.01). Hydroxyproline contents was reduced obviously(P<0.01). Apoptotic rate and expressions of Fas/FasL were decreased significantly(P<0.01). Conclusion:Pulmonary fibrosis was improved significantly after the endogenous IL-17 activity blocked,meanwhile,apoptosis and the expressions of Fas/FasL were both decreased. These data showed that antimouse IL-17 mAb ameliorate the pulmonary fibrosis induced by BLM and it is related to Fas/FasL apoptotic pathways.

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张效云,宋桂芹,辇晓峰,黄 勇,郝 敏,孙 冲.阻断白介素-17对博来霉素诱导小鼠肺纤维化及肺组织Fas/FasL表达的影响[J].南京医科大学学报(自然科学版),2017,(5):584-587,624

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  • 收稿日期:2016-10-27
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  • 在线发布日期: 2017-06-01
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