中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (50): 9453-9457.doi: 10.3969/j.issn.2095-4344.2012.50.026

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

人肺和支气管成纤维细胞骨架蛋白的表达和增殖

方秋红1,税朝祥2,王尧尧2,王瑞琴2,王  晶1,马迎民1   

  • 出版日期:2012-12-09 发布日期:2012-12-09

Expression and proliferation of human lung and bronchial fibroblast cytoskeleton protein

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

  • Online:2012-12-09 Published:2012-12-09

摘要:

背景:假设支气管和肺成纤维细胞在损伤修复过程中可能表现出不同的生物学特性,从而在慢性阻塞性肺病气道炎症过程中,支气管和肺组织呈现出不同的重构特点。
目的:在体外培养的人肺和支气管成纤维细胞上,观察骨架蛋白的表达以及二者增殖特性的差别,以助阐明慢性阻塞性肺病支气管和肺组织重构的发生机制。
方法:采用人肺和支气管成纤维细胞体外培养;免疫组织化学方法测定细胞波形蛋白和肌动蛋白的表达;MTT法测定细胞增殖。
结果与结论:人肺和支气管成纤维细胞均显著表达波形蛋白和肌动蛋白,但两种蛋白在细胞内的分布不同。波形蛋白呈斑点状分布于肺成纤维细胞胞浆内,并以细胞核周围为主;在支气管成纤维细胞内,波形蛋白均匀分布于细胞浆内。在肺成纤维细胞,肌动蛋白沿细胞膜分布;在支气管成纤维细胞,肌动蛋白广泛分布于细胞浆。在相同的培养条件下,支气管和肺组织成纤维细胞的增殖速度不同,支气管成纤维细胞增殖较肺成纤维细胞增殖速度显著增加。提示支气管和肺成纤维细胞增殖修复特性不同,这些生物学特性的差别可能在慢性阻塞性肺病支气管和肺组织不同的重构过程中发挥重要作用。

Abstract:

BACKGROUND: The bronchial and lung tissue shows different reconstruction characteristics in the process of chronic obstructive pulmonary disease as the bronchial and lung fibroblasts may exhibit different biological characteristics during the process for repairing defects. 
OBJECTIVE: To observe the cytoskeleton protein expression and the proliferation of human lung and bronchial fibroblasts cultured in vitro in order to clarify the mechanism of bronchial and lung tissue remodeling of chronic obstructive pulmonary disease. 
METHODS: Human lung and bronchial fibroblasts were cultured in vitro; the expression of vimentin and α smooth muscle actin was detected by immunohistochemistry; MTT assay was utilized for measuring the fibroblasts proliferation.
RESULTS AND CONCLUSION: Vimentin and α smooth muscle actin were intensely expressed in the human lung and bronchial fibroblasts, but the distribution was different between lung and bronchial fibroblasts. In the cytoplasm of lung fibroblasts, vimentin was expressed mainly around the nuclear in a dot-like pattern, and vimentin was distributed along the cell membrane. In the bronchial fibroblasts, the vimentin was distributed along the cell membrane; in the lung fibroblasts, α smooth muscle actin was distributed in the cell membrane. Under the same culture condition, the proliferative degree of lung and bronchial fibroblasts was different. The bronchial fibroblasts proliferated significantly faster than the lung fibroblasts. These findings suggest that bronchial and lung fibroblasts behave differently in the repair and regenerative process of lung and bronchial tissue remodeling, which might play a pivotal role in the airflow limitation of chronic obstructive pulmonary disease.