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Enhanced expression of transglutaminase 2 in anterior polar cataracts and its induction by TGF-β in vitro

机译:转谷氨酰胺酶2在前极性白内障中的增强表达及其在体外的TGF-β诱导

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摘要

>Background/aims: Transglutaminase activity has long been implicated in the cataract formation. However, the precise mechanism of how it is produced and involved in this process remains unclear. Here the authors sought to examine whether transglutaminase 2 (TGase 2) is expressed in lens epithelial cells from patients with anterior polar cataracts, to determine whether TGase 2 expression is induced by transforming growth factor (TGF-β) in cultured lens epithelial cells, and to determine whether TGase 2 participates in the crosslinking of fibronectin in lens epithelial cells in vitro.>Methods: Lens epithelial cells from anterior polar cataracts, nuclear cataracts, and non-cataractous clear lenses were examined for the expression of TGase 2 using reverse transcription-polymerase chain reaction, western blot analysis, and immunohistochemical analysis. The modulation of extracellular TGase 2 activity by TGF-β was measured by the formation of fibronectin polymers and the incorporation of fluorescein cadaverine into extracellular matrix proteins. The effect of TGase 2 overexpression was analysed by immunofluorescence staining and western blot analysis of human lens epithelial (HLE) B-3 cells transiently transfected with TGase 2 gene.>Results: The expression of TGase 2 mRNA and its protein was markedly enhanced in lens epithelial cells from patients with anterior polar cataracts. Treatment of HLE B-3 cells with TGF-β caused an increase in TGase 2 protein, its extracellular activity, and the crosslinking of fibronectin. Transient transfection of HLE B-3 cells with the TGase 2 gene led to the increased production of fibronectin monomers and polymers.>Conclusions: This study shows that TGase 2 is overexpressed in lens epithelial cells from anterior polar cataracts and that TGF-β may be a causative factor in the induction of TGase 2. The enhanced expression of TGase 2 might cause the accumulation and crosslinking of the extracellular matrix proteins and might play a part in anterior polar cataract development.
机译:>背景/目标:转谷氨酰胺酶活性早已与白内障形成有关。但是,如何生产和参与该过程的确切机制仍不清楚。在这里,作者试图检查转谷氨酰胺酶2(TGase 2)在患有前极性白内障患者的晶状体上皮细胞中是否表达,以确定在培养的晶状体上皮细胞中是否由转化生长因子(TGF-β)诱导了TGase 2的表达,以及以确定TGase 2是否参与体外晶状体上皮细胞中纤连蛋白的交联。>方法:检查前极性白内障,核性白内障和非白内障透明晶状体的晶状体上皮细胞的表达TGase 2使用逆转录-聚合酶链反应,蛋白质印迹分析和免疫组化分析。通过形成纤连蛋白聚合物和将荧光素尸胺掺入细胞外基质蛋白来测量TGF-β对细胞外TGase 2活性的调节。通过免疫荧光染色和蛋白质印迹分析了瞬时转染了TGase 2基因的人晶状体上皮细胞(HLE)B-3细胞中TGase 2的表达。>结果:前极性白内障患者晶状体上皮细胞中的蛋白质显着增强。用TGF-β处理HLE B-3细胞可导致TGase 2蛋白,其细胞外活性和纤连蛋白交联增加。用TGase 2基因瞬时转染HLE B-3细胞导致纤连蛋白单体和聚合物的产量增加。>结论:该研究表明,前极性白内障和白内障晶状体上皮细胞中TGase 2过表达。 TGF-β可能是诱导TGase 2的诱因。TGase2的表达增强可能引起细胞外基质蛋白的积累和交联,并可能参与前极性白内障的发展。

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