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羧甲基殼聚糖及羧甲基甲殼素對TGF-β誘導(dǎo)的角膜上皮纖維化的抑制作用研究

發(fā)布時間:2018-08-15 15:26
【摘要】:本實驗旨在研究不同分子量的羧甲基殼聚糖(CMCTS)和羧甲基甲殼素(CMCT)對轉(zhuǎn)化生長因子β(TGF-β)誘導(dǎo)的人角膜上皮細胞纖維化的抑制作用,并初探其作用機制。MTT法檢測不同分子量的CMCTS和CMCT對人角膜上皮細胞系(HCE)生長增殖能力的影響;細胞劃痕實驗研究TGF-β誘導(dǎo)的HCE細胞纖維化過程中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對角膜上皮細胞纖維化相關(guān)的TGF-β/Smad通路信號分子的影響。結(jié)果表明,在10~1 000μg/mL濃度范圍內(nèi),4種多糖對HCE細胞均具有較好的相容性。此外,不論大分子還是小分子的CMCTS對TGF-β誘導(dǎo)的HCE細胞纖維化均具有顯著的抑制作用,其作用機制與抑制TGF-β/Smad信號通路Smad2和Smad3分子的磷酸化有關(guān)。然而,兩種分子量的CMCT均沒有抑制纖維化的作用。因此,CMCTS可以作為一種有發(fā)展?jié)摿Φ纳镝t(yī)用材料應(yīng)用于人角膜上皮損傷修復(fù),發(fā)揮其抗纖維化的作用。
[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.
【作者單位】: 青島大學(xué)醫(yī)學(xué)院;青島大學(xué)附屬醫(yī)院中心實驗室;
【基金】:山東省科技發(fā)展計劃項目(2014GHY115025) 山東省自然科學(xué)基金培養(yǎng)基金項目(ZR2014HP011)資助~~
【分類號】:R772.2

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