中国组织工程研究

中国组织工程研究 ›› 2013, Vol. 17 ›› Issue (11): 2020-2025.doi: 10.3969/j.issn.2095-4344.2013.11.018

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

胰蛋白酶与糖皮质激素影响肺成纤维细胞增殖与骨架蛋白的表达

方秋红1，税朝祥2，王尧尧2，王瑞芹2，王 晶1，马迎民1   

  1. 1首都医科大学附属北京世纪坛医院呼吸与危重医学科，北京市 100038
    2 清华大学第一附属医院，北京市 100016
  • 收稿日期:2012-07-01 修回日期:2012-08-24 出版日期:2013-03-12 发布日期:2013-03-12
  • 通讯作者: 马迎民，博士，主任医师，教授，首都医科大学附属北京世纪坛医院呼吸与危重医学科，北京市 100038 sjthxm@yahoo.cn
  • 作者简介:方秋红☆，女，1968年生，河北省唐山市人，汉族，1998年北京大学医学部毕业，博士，主任医师，主要从事慢性阻塞性肺病组织损伤修复机制的研究。 florence408@126.com

Trypsin and dexamethasone affect proliferation and cytoskeleton protein expression of human lung fibroblasts

Fang Qiu-hong1, Shui Chao-xiang2, Wang Yao-yao2, Wang Rui-qin2, Wang Jing1, Ma Ying-min1   

  1. 1 Department of Pulmonary and Critical Care, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China
    2 First Hospital of Tsinghua University, Beijing 100016, China
  • Received:2012-07-01 Revised:2012-08-24 Online:2013-03-12 Published:2013-03-12
  • Contact: Ma Ying-min, Doctor, Chief physician, Professor, Department of Pulmonary and Critical Care, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China sjthxm@yahoo.cn
  • About author:Fang Qiu-hong☆, Doctor, Chief physician, Department of Pulmonary and Critical Care, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China florence408@126.com

PDF

554

被引次数

1

摘要：

背景：肺成纤维细胞不但是肺组织的结构支持细胞，还能够通过增殖、收缩、趋化及分泌细胞外基质等多种功能在肺组织的炎症损伤-修复过程中发挥关键作用。
目的：观察原代培养的人肺成纤维细胞在胰蛋白酶或糖皮质激素作用下增殖特性与骨架蛋白表达的变化。
方法：采用人肺成纤维细胞体外培养，胰酶处理24 h，终浓度分别为0，0.5，1.0，5，10 mg/L胰酶；地塞米松处理72 h，在有血清培养条件下进行，浓度范围10-9-10-6 mol/L。MTT法测定成纤维细胞增殖情况；免疫印迹方法测定细胞波形蛋白和肌动蛋白的表达。
结果与结论：胰蛋白酶在低浓度(0.1-0.5 mg/L)时刺激肺成纤维细胞增殖；而较高浓度(1-10 mg/L)明显抑制细胞生长。地塞米松对肺成纤维细胞增殖作用的影响不明显。胰蛋白酶显著上调肺成纤维细胞α平滑肌肌动蛋白的表达，但对波形蛋白的表达无明显影响；地塞米松(10-9-10-6 mol/L)显著抑制波形蛋白的表达。结果表明在慢性阻塞性肺病等支气管肺慢炎症性疾病的发病过程中，胰蛋白酶和糖皮质激素可以通过参与成纤维细胞增殖和骨架蛋白的表达发挥作用。

关键词: 组织构建, 组织构建细胞学实验, 胰蛋白酶, 糖皮质激素, 肺成纤维细胞, 波形蛋白, 肌动蛋白, 免疫印迹, 地塞米松\骨架蛋白, 成纤维细胞增殖, 慢性阻塞性肺病, 国家自然科学基金, 组织构建图片文章

Abstract:

BACKGROUND: In addition to the supporting function, lung fibroblasts play a pivotal role in the process of lung tissue injury and repair through proliferation, contraction, chemotaxis and secretion of the extracellular matrix.
OBJECTIVE: To investigate the effects of trypsin and dexamethasone on the proliferation and cytoskeleton protein expression of lung fibroblasts.
METHODS: Human lung fibroblasts were isolated and cultured in vitro followed by trypsin (0, 0.5, 1.0, 5, 10 mg/L) treatment for 24 hours or dexamethasone (10-9-10-6 mol/L) treatment for 72 hours. The expression of vimentin and α-smooth muscle actin was detected by immunoblotting. 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay was utilized for measuring fibroblasts proliferation.
RESULTS AND CONCLUSION: Trypsin at lower concentrations (0.1-0.5 mg/L) significantly stimulated lung fibroblasts proliferation, while higher concentrations (1-10 mg/L) of trypsin inhibited lung fibroblasts proliferation. No effect on fibroblasts proliferation was observed with dexamethasone. Trypsin significantly increased the expression of α-smooth muscle actin in a dose-dependent manner, but had no influence on vimentin expression. Dexamethasone (10-9-10-6 mol/L) significantly inhibited vimentin expression in a concentration-dependent manner. The data reveal that trypsin and glucocorticosteroids may participate in the tissue remodeling process of chronic inflammatory airway diseases through affecting lung fibroblasts proliferation and the expression of cytoskeleton protein.

Key words: tissue construction, cytology experiment in tissue construction, trypsin, glucocorticoids, lung fibroblasts, vimentin, actin, immunoblotting, dexamethasone, cytoskeletal proteins, fibroblast proliferation, chronic obstructive pulmonary disease, the National Natural Science Foundation of China, tissue construction photographs-containing paper

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