周海艳,李静云,殷松楼.结缔组织病相关肺间质病变肺组织中TIMP-1表达及其TGF-β1/Smad3途径调控作用分析.[J].中南医学科学杂志.,2019,(6):628-632.
结缔组织病相关肺间质病变肺组织中TIMP-1表达及其TGF-β1/Smad3途径调控作用分析
Expression of TIMP-1 in lung tissues of connective tissue disease-related pulmonary interstitial lesions and the role of related regulatory factors
投稿时间:2019-04-18  修订日期:2019-09-16
DOI:10.15972/j.cnki.43-1509/r.2019.06.017
中文关键词:  结缔组织病  肺间质病变  人肺成纤维细胞  TIMP-1  TGF-β1
英文关键词:connective tissue disease  pulmonary interstitial disease  human lung fibroblasts  TIMP-1  TGF-β1
基金项目:
作者单位
周海艳 徐州医科大学研究生学院风湿科,江苏 徐州 223000 
李静云 徐州医科大学附属沭阳医院风湿科, 江苏 徐州223600 
殷松楼 徐州医科大学附属医院风湿免疫科,江苏 徐州223000 
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中文摘要:
      探讨结缔组织病相关肺间质病变肺组织中基质金属蛋白酶组织抑制因子(TIMP)-1表达水平。收集结缔组织病相关肺间质病变患者行CT引导下经皮肺穿刺活检时的肺组织(观察组),以及无感染和吸烟史的肺癌患者远离癌症病灶的正常肺组织(对照组),行TIMP-1免疫荧光染色分析。原代培养人肺成纤维细胞,选用第10~15代的细胞,以不同浓度TGF-β1诱导24 h,以及终浓度为10 μg/L的TGF-β1诱导不同时间。实验结果表明,观察组肺组织中TIMP-1荧光强度明显高于对照组(P<0.05)。TGF-β1浓度3、5、10、15 μg/L作用于人肺成纤维细胞24 h后,TIMP-1 mRNA和蛋白相对表达量明显高于0 μg/L(P<0.05)。使用10 μg/L TGF-β1作用24、48 h时,TIMP-1 mRNA和蛋白相对表达量明显高于作用0 h时(P<0.05)。Smad3真核过表达载体组TIMP-1、P300 mRNA和蛋白相对表达量明显高于空质粒转染组、空白对照组(P<0.05)。实验证明,结缔组织病相关肺间质病变肺组织中TIMP-1表达明显提升,TGF-β1可通过TGF-β1/Smad3信号通路上调TIMP-1表达,参与结缔组织病相关肺间质病变的发生与发展。
英文摘要:
      To investigate the expression of tissue inhibitor of matrix metalloproteinase (TIMP) -1 and the role of related regulatory factors in lung tissue of connective tissue disease-related pulmonary interstitial lesions. The lung tissues of patients with connective tissue disease-related pulmonary interstitial lesions undergoing CT-guided percutaneous lung biopsy (observation group) and normal lung tissues (control group) of lung cancer patients with no history of infection and smoking were collected and analyzed by TIMP-1 immunofluorescence staining. Human lung fibroblasts were cultured in primary culture. The cells of the 10th to 15th generations were induced by different concentrations of TGF-β1 for 24 hours, and the final concentration of 10 μg/L was induced by TGF-β1 for different time.The results showed that the fluorescence intensity of TIMP-1 in the lung tissue of the observation group was significantly higher than that of the control group (P<0.05). The relative expression of TIMP-1 protein and TIMP-1 mRNA was significantly higher than that of 0 μg/L after the treatment of human lung fibroblasts with 3,5, 10 and 15 μg/L of TGF-β1 for 24 hours (P<0.05). When treated with 10 μg/L TGF-β1 for 24 h and 48 h, the relative expression of TIMP-1 protein and TIMP-1 mRNA was significantly higher than that at 0 h (P<0.05). The TIMP-1,P300 and protein relative expressions of Smad3 eukaryotic overexpression vector group were significantly higher than those in blank plasmid transfection group and blank control group (P<0.05). So The expression of TIMP-1 in lung tissues of connective tissue disease-related pulmonary interstitial lesions is significantly increased, and the expression of TIMP-1 can be up-regulated by TGF-β1,Smad3 participate in the occurrence and development of connective tissue disease-related pulmonary interstitial lesions through the TGF-β1/Smad3 signaling pathway.
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