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  

Inhibitory Effect of CMCTS or CMCT on Corneal Epithelial Fibrosis Induced by TGF-β

作　　者：王哲颖[1] 隋爱华[2] 徐文华[1] 刘成玉[1] 周泉[2] 王丽萍[2] 梁晔[2] 

机构地区：[1]青岛大学医学院,山东青岛266003 [2]青岛大学附属医院中心实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2017年第7期96-104,共9页Periodical of Ocean University of China

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~~

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。In this study, carboxymethyl chitosan （CMCTS） and carboxymethyl chitin （CMCT） with different molecular weights were explored on their anti-fibrosis effect and the affecting mechanism. The effects of different kinds of CMCTS and CMCT on the growth of the human corneal epithelial cell line （HCE） were measured by MTT assay, while their anti-fibrosis effects on the migration of HCE ceils were reflected by wound healing assay. The anti-fibrosis mechanism of different kinds of CMCTS and CMCT in corneal epithelia reconstruction was further studied on protein level. Results showed that CM- CTS and CMCT with different molecular weights all suited the proliferation of HCE cells within the concentration of 10-1 000 μg/mL. But the two kinds of CMCTS could exert inhibitory effect on the fi- brosis of HCE cells induced by TGF-β, and the anti-fibrosis mechanism was related to reducing phos- phorylation of Smad2 and Smad3 protein in the TGF-β/Smad signaling pathway. Therefore, the two kinds of CMCTS could be applied in the repair of human corneal epithelial injury, playing an important role in anti-fibrosis.

关 键 词：羧甲基壳聚糖 角膜上皮 纤维化 TGF-Β 

分 类 号：Q539.7[生物学—生物化学]