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重组人血管生成素-1与肺间质纤维化血管正常化时相的关系

  

  1. 1.河北省胸科医院 河北省肺癌防治中心 a.结核科;b.病理科;c.生物治疗科,河北 石家庄 050041;
    2.河北师范大学  生命科学学院,河北 石家庄 050024
  • 出版日期:2016-09-05 发布日期:2016-08-31
  • 通讯作者: 通信作者:杨永辉,Email:yonghuiyang@vip.163.com
  • 基金资助:
    国家自然科学基金资助项目(31101638);河北省省级重大医学研究课题(ZD2013063);河北省省级重大医学研究
    课题(ZD2013060);河北省级科技计划项目(14277789D)

Relationship between human angiopoietin1 and pulmonary fibrosis

  1. 1a.Department of Tuberculosis;1b.Department of Pathology;1c.Department of Biological Therapy,
    Hebei Chest  Hospital, Shijiazhuang 050041, China;2.College of  Life Science,
    Hebei Normal  University, Shijiazhuang, 050024, China
  • Online:2016-09-05 Published:2016-08-31
  • Contact: Corresponding author: Yang Yonghui, Email:yonghuiyang@vip.163.com

摘要: 目的探讨重组人血管生成素1(Human Angiopoietin1,rHuANG1 )对小鼠模型肺间质纤维化微血管正常化的最佳时相及其分子机制。方法将40只C57/BL6小鼠按掷硬币法随机分成对照组和实验组,每组各20只。对照组腹腔注射生理盐水(NS)(0.2 ml/d);实验组腹腔注射rHuANG1(5 mg·kg-1·d-1)。每组分别于治疗后的第2、4、6、9天处死5只小鼠留取标本。测量小鼠肺组织羟脯氨酸含量变化。酶联免疫吸附测定法(ELISA)检测肺组织中G蛋白调节信号5 (regulator of Gprotein signaling 5, RGS5)蛋白浓度,苏木精伊红(HE)染色检测肺组织病理变化,免疫组织化学形态学分析血管内皮生长因子(vascular endothelial growth factor, VEGF)的变化。结果①成功建立C57/BL6小鼠肺间质纤维化模型。②实验组接受rHuANG1注射后,第6天,第9天肺羟脯氨酸含量降低(P<0.01)。③ ELISA结果显示,实验组肺组织中RGS5的表达在治疗第4天和第6天与对照组比较明显增高(P<0.05)。④HE染色结果显示,实验组和对照组不同时间点均可见不同程度的肺纤维化。对照组坏死程度高于实验组。其中,实验组第4天和第6天组织修复最明显,而对照组不同时点肺间质纤维化比较明显。⑤免疫组织化学结果表明,与对照组相比,实验组第4、6天VEGF在肺间质纤维化中的表达率升高明显减少(P<0.05)。结论rHuANG1作用于小鼠肺间质纤维化后第4~6天可作为血管正常化时间窗,可能与VEGF相关。

关键词: 肺纤维化, 血管生成素1, G蛋白调节信号5, 血管内皮生长因子

Abstract: ObjectiveTo analyze the optimal time course and relevant molecular mechanism of recombinant human angiopoietin1 (rHuANG1) in the treatment of pulmonary fibrosis in mice. MethodsThe model of pulmonary fibrosis was established on C57/BL6 mice. And 40 mice were subdivided into NS group and rHuANG1 group (20 mice per group). NS group experienced treatment of intraperitoneal injection NS  0.2 ml/d, while rHuANG1 group was treated rHuANG1 for 5 mg·kg-1·d-1. The samples of 5 mice were obtained at d2,d4,d6 and d9 after treatment in NS group and rHuANG1 group, respectively. Regulator of Gprotein signaling 5(RGS5) was estimated by ELISA in lung. Results①The pulmonary fibrosis model on C57/BL6 mice was established successfully. ②Hydroxyprolinc in the lung of NS group and rHuANG1 group at d6  and d9 reduced significantly(P<0.01). ③ ELISA results indicated the expression of RGS5 in the lung of rHuANG1 group at d4 and d6 increased as compared with that of NS group(P<0.05). ④HE staining results showed that different degrees of pulmonary fibrosis were observed at different periods. The degree of necrosis in NS group was higher than that of rHuANG1 group. Among them, tissue repair in rHuANG1 group was more obvious at d4 and d6, while NS group did not present the same result at the corresponding time. ⑤The results of immunohistochemistry assay showed that the expression of VEGF in rHuANG1 group had significalnt decrease at d4 and d6 as compared with that of in NS group(P<0.05).ConclusionrHuANG1 could have positive effect on pulmonary fibrosis model of C57/BL6 mice. The time d4d6 was considered as a brief normalized time window due to the normalization of VEGF.

Key words: pulmonary fibrosis, human angiopoietin1, regulator of Gprotein signaling 5, vascular endothelial growth factors