羧甲基壳聚糖及羧甲基甲壳素对TGF-β诱导的角膜上皮纤维化的抑制作用研究

发布时间：2018-08-15 15:26

【摘要】：本实验旨在研究不同分子量的羧甲基壳聚糖(CMCTS)和羧甲基甲壳素(CMCT)对转化生长因子β(TGF-β)诱导的人角膜上皮细胞纤维化的抑制作用,并初探其作用机制。MTT法检测不同分子量的CMCTS和CMCT对人角膜上皮细胞系(HCE)生长增殖能力的影响;细胞划痕实验研究TGF-β诱导的HCE细胞纤维化过程中CMCTS和CMCT的作用;western blot方法在蛋白水平探究不同分子量的CMCTS和CMCT对角膜上皮细胞纤维化相关的TGF-β/Smad通路信号分子的影响。结果表明,在10~1 000μg/mL浓度范围内,4种多糖对HCE细胞均具有较好的相容性。此外,不论大分子还是小分子的CMCTS对TGF-β诱导的HCE细胞纤维化均具有显著的抑制作用,其作用机制与抑制TGF-β/Smad信号通路Smad2和Smad3分子的磷酸化有关。然而,两种分子量的CMCT均没有抑制纤维化的作用。因此,CMCTS可以作为一种有发展潜力的生物医用材料应用于人角膜上皮损伤修复,发挥其抗纤维化的作用。
[Abstract]:The aim of this study was to investigate the inhibitory effects of carboxymethyl chitosan (CMCTS) and carboxymethyl chitin (CMCT) on transforming growth factor 尾 (TGF- 尾) -induced fibrosis in human corneal epithelial cells. The effects of different molecular weight of CMCTS and CMCT on the growth and proliferation of human corneal epithelial cell line (HCE) were studied. The role of CMCTS and CMCT in the process of HCE cell fibrosis induced by TGF- 尾 was investigated by cell scratch assay. The effects of CMCTS and CMCT with different molecular weights on the signal molecules of TGF- 尾 / Smad pathway associated with corneal epithelial fibrosis were investigated at the protein level by western blot method. The results showed that the four polysaccharides had good compatibility with HCE cells in the concentration range of 10 ~ 1 000 渭 g/mL. In addition, CMCTS of both macromolecules and small molecules significantly inhibited the fibrosis of HCE cells induced by TGF- 尾, and its mechanism was related to the inhibition of phosphorylation of Smad2 and Smad3 molecules in TGF- 尾 / Smad signaling pathway. However, CMCT with both molecular weights did not inhibit fibrosis. Therefore CMCTS can be used as a potential biomedical material for the repair of human corneal epithelial injury and play its role in anti-fibrosis.
【作者单位】：青岛大学医学院;青岛大学附属医院中心实验室;
【基金】：山东省科技发展计划项目(2014GHY115025) 山东省自然科学基金培养基金项目(ZR2014HP011)资助~~
【分类号】：R772.2

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