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原代培养鼠肺成纤维细胞中α-平滑肌肌动蛋白的表达
作者姓名:Yang JP  Su DJ  Li SN  Gao L
作者单位:内蒙古医学院第三附属医院呼吸与危重症科,内蒙古包头014010
基金项目:内蒙古医学院科技研究重大项目资助(NY200SZD007)
摘    要:目的:观察原代培养大鼠肺成纤维细胞(FB)中α-平滑肌肌动蛋白(α-SMA)的表达情况。方法:24只雌性Wistar大鼠,随机分为正常组和模型组,气管内注入博莱霉素A2(BLM-A2)(5mg/kg)建立大鼠肺间质纤维化(PF)模型,正常组气管内注入等量生理盐水。实验周期第28d腹主动脉放血法处死,肺组织HE染色和透射电镜检查。体外原代培养FB,传3-5代后流式细胞术检测α-SMA表达的细胞阳性率。结果:HE染色28d模型组大片肺泡结构破坏,肺泡腔消失,有成纤维细胞灶形成,电镜观察肺组织超微结构发生改变,肺间质中可见胞浆内有微丝样结构的MF。流式细胞分析结果:正常组α-SMA表达的细胞阳性率为95.5%±4.68%,模型组α-SMA表达的细胞阳性率为96.2%±2.14%,与正常组相比P〉0.05。结论:原代培养的大鼠肺FB中α-SMA的阳性表达率正常组和模型组均显著增高,体外培养的大鼠肺FB传3—5代后可能有部分已经转化为肌纤维母细胞(MF)。

关 键 词:α-SMA  成纤维细胞  原代培养  表型转化

The expression of alpha-smooth muscle actin in primary cultural fibroblasts of rats
Yang JP,Su DJ,Li SN,Gao L.The expression of alpha-smooth muscle actin in primary cultural fibroblasts of rats[J].Chinese Journal of Applied Physiology,2009,25(3):339-343.
Authors:Yang Jing-ping  Su De-jun  Li Shu-nan  Gao Li
Affiliation:Department of Respiratory and Critical Care Medicine, The Third Affiliated Hospital, Inner Mongolia Medical College, Baotou 014010, China. yangron@sina.com.cn
Abstract:Aim: To observe the expression of α-smooth muscle actin(a-SMA) in primary cultural fibroblasts of rats. Methods: 12 female Wistar rats were randomly assigned into two groups, the normal group and the model group. The model group was fdled with bleomycin A2(5mg/kg)once into the trachea. The normal group was filled with equal saline into the trachea. The rats were sacrificed under drugged state at 28 days of feeding, then Hematoxylin-Eosin staining and electron microscopy were used to evaluate the foundation of the model. The isolated fibroblasts from the rats were cultured in vitro. Flow cytometry was used in the test to observe the expression of a-SMA in fibroblast in vitro in rats. Resuits: The formation of fibroblast loci was observed in the model group by optical tnicroscope. The ultrastructure in pulmonary tissue of the model group rats were changed and proliferated myofibroblasts with filaments were found in the alveolar septa by electron microscopy. The expression of α-SMA was positive in the normal and model group. There was no difference between the two groups in the rates of positive cells( P 〉 0.05). Conclusion: Both the normal and model groups had the phenotype conversion in lung fibroblasts in vitro.
Keywords:α-SMA  primary culture  lung fibroblasts  phenotype conversion
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