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转化生长因子-β2在体外诱导人角膜内皮细胞发生形态改变。

Transforming growth factor-β2 induces morphological alteration of human corneal endothelial cells in vitro.

作者信息

Wang Jing, Fan Ting-Jun, Yang Xiu-Xia, Chang Shi-Min

机构信息

College of Life Sciences, Langfang Normal University, Langfang 065000, Hebei Province, China.

Key Laboratory for Corneal Tissue Engineering, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, Shandong Province, China.

出版信息

Int J Ophthalmol. 2014 Oct 18;7(5):759-63. doi: 10.3980/j.issn.2222-3959.2014.05.03. eCollection 2014.

Abstract

AIM

To investigate the morphological altering effect of transforming growth factor-β2 (TGF-β2) on untransfected human corneal endothelial cells (HCECs) in vitro.

METHODS

After untransfected HCECs were treated with TGF-β2 at different concentrations, the morphology, cytoskeleton distribution, and type IV collagen expression of the cells were examined with inverted contrast light microscopy, fluorescence microscopy, immunofluorescence or Western Blot.

RESULTS

TGF-β2 at the concentration of 3-15 µg/L had obviously alterative effects on HCECs morphology in dose and time-dependent manner, and 9 µg/L was the peak concentration. TGF-β2 (9 µg/L) altered HCE cell morphology after treatment for 36h, increased the mean optical density (P<0.01) and the length of F-actin, reduced the mean optical density (P<0.01) of the collagen type IV in extracellular matrix (ECM) and induced the rearrangement of F-actin, microtubule in cytoplasm and collagen type IV in ECM after treatment for 72h.

CONCLUTION

TGF-β2 has obviously alterative effect on the morphology of HCECs from polygonal phenotype to enlarged spindle-shaped phenotype, in dose and time-dependence manner by inducing more, elongation and alignment of F-actin, rearrangement of microtubule and larger spread area of collagen type IV.

摘要

目的

研究转化生长因子-β2(TGF-β2)对体外未转染的人角膜内皮细胞(HCECs)形态的改变作用。

方法

用不同浓度的TGF-β2处理未转染的HCECs后,采用倒置相差显微镜、荧光显微镜、免疫荧光或蛋白质免疫印迹法检测细胞的形态、细胞骨架分布及IV型胶原表达。

结果

3-15μg/L的TGF-β2对HCECs形态有明显的剂量和时间依赖性改变作用,9μg/L为峰值浓度。9μg/L的TGF-β2处理36小时后可改变HCE细胞形态,增加F-肌动蛋白的平均光密度(P<0.01)和长度,处理72小时后降低细胞外基质(ECM)中IV型胶原的平均光密度(P<0.01),并诱导F-肌动蛋白、细胞质中的微管及ECM中的IV型胶原重排。

结论

TGF-β2对HCECs形态有明显的改变作用,使其从多边形表型转变为增大的纺锤形表型,呈剂量和时间依赖性,通过诱导更多、更长且排列整齐的F-肌动蛋白、微管重排以及更大面积的IV型胶原铺展来实现。

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