Kwok Sum Sum, Wong Francisca Siu-Yin, Shih Kendrick Co, Chan Yau-Kei, Bu Yashan, Chan Tommy Chung-Yan, Ng Alex Lap-Ki, Lo Amy Cheuk-Yin, Tong Louis, Yam Gary Hin-Fai, Jhanji Vishal
Department of Ophthalmology, Li Ka Shing Faculty of Medicine, University of Hong Kong and Hong Kong SAR, Hong Kong, China.
Cornea and External Eye Disease Service, Singapore National Eye Centre, Singapore 168751, Singapore.
J Clin Med. 2020 Nov 6;9(11):3572. doi: 10.3390/jcm9113572.
(1) Objective: To study the anti-fibrotic effects of polysaccharides (LBP) on corneal stromal fibroblasts and assess LBP's effect on cell viability. (2) Methods: Primary human corneal keratocytes of passage 3 to 6 were used for all experiments. Cells are pretreated with LBP solution for 24 h and then transforming growth factor beta 1 (TGFβ1) for 48 h and collected for experiments. Fibrotic protein analysis was performed using immunofluorescence and Western blot. The effect of LBP on cell viability was assessed using the MTS assay. (3) Results: LBP significantly reduced the expression of fibrotic proteins, including α-SMA and extracellular matrix proteins (collagen type I and III). LBP significantly decreased the viability of myofibroblasts but not the fibroblasts. Conclusions: In this study, LBP was effective in the prevention of fibrosis gene expression. Further studies to assess the underlying mechanism and pharmacological properties will facilitate the formation of a topical LBP solution for in vivo studies.
(1) 目的：研究枸杞多糖（LBP）对角膜基质成纤维细胞的抗纤维化作用，并评估LBP对细胞活力的影响。(2) 方法：所有实验均使用第3至6代原代人角膜基质细胞。细胞先用LBP溶液预处理24小时，然后用转化生长因子β1（TGFβ1）处理48小时，收集细胞用于实验。使用免疫荧光和蛋白质印迹法进行纤维化蛋白分析。使用MTS法评估LBP对细胞活力的影响。(3) 结果：LBP显著降低了纤维化蛋白的表达，包括α-平滑肌肌动蛋白（α-SMA）和细胞外基质蛋白（I型和III型胶原）。LBP显著降低了肌成纤维细胞的活力，但对成纤维细胞没有影响。结论：在本研究中，LBP可有效预防纤维化基因表达。进一步评估其潜在机制和药理特性的研究将有助于形成用于体内研究的局部LBP溶液。
