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枸杞多糖对角膜上皮-基质损伤的抗瘢痕作用。

The anti-scarring role of Lycium barbarum polysaccharide on cornea epithelial-stromal injury.

机构信息

Department of Ophthalmology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region.

Department of Mechanical Engineering, Faculty of Engineering, The University of Hong Kong, Hong Kong Special Administrative Region.

出版信息

Exp Eye Res. 2021 Oct;211:108747. doi: 10.1016/j.exer.2021.108747. Epub 2021 Aug 24.

Abstract

PURPOSE

Cornea epithelial-stromal scarring is related to the differentiation of fibroblasts into opaque myofibroblasts. Our study aims to assess the effectiveness of Lycium barbarum polysaccharide (LBP) solution as a pre-treatment in minimizing corneal scarring.

METHODS

Human corneal fibroblasts were cultured in a three-dimensional collagen type I-based hydrogel in an eye-on-a-chip model. Fibroblasts were pre-treated with 2 mg/mL LBP for 24 h, followed by another 24-h incubation with 10 ng/mL transforming growth factor-beta 1 (TGF-β1) to induce relevant physiological events after stromal injury. Intracellular pro-fibrotic proteins, extracellular matrix proteins, and pro-inflammatory cytokines that involved in fibrosis, were assessed using immunocytochemistry and enzyme-linked immunosorbent assays.

RESULTS

Compared to the positive control TGF-β1 group, LBP pre-treated cells had a significantly lower expression of alpha-smooth muscle actin, marker of myofibroblasts, vimentin (p < 0.05), and also extracellular matrix proteins both collagen type II and type III (p < 0.05) that can be found in scar tissues. Moreover, LBP pre-treated cells had a significantly lower secretion of pro-inflammatory cytokines interleukin-6 and interleukin-8 (p < 0.05). The cell-laden hydrogel contraction and stiffness showed no significant difference between LBP pre-treatment and control groups. Fibroblasts pretreated with LBP as well had reduced angiogenic factors expression and suppression of undesired proliferation (p < 0.05).

CONCLUSION

Our results showed that LBP reduced both pro-fibrotic proteins and pro-inflammatory cytokines on corneal injury in vitro. We suggest that LBP, as a natural Traditional Chinese Medicine, may potentially be a novel topical pre-treatment option prior to corneal refractive surgeries with an improved prognosis.

摘要

目的

角膜上皮-基质瘢痕与成纤维细胞分化为不透明的肌成纤维细胞有关。本研究旨在评估枸杞多糖(LBP)溶液作为预处理剂以最小化角膜瘢痕的有效性。

方法

在眼芯片模型中,用人角膜成纤维细胞在三维胶原 I 基水凝胶中培养。将成纤维细胞用 2mg/ml LBP 预处理 24 小时,然后再用 10ng/ml 转化生长因子-β 1(TGF-β1)孵育 24 小时,以诱导基质损伤后的相关生理事件。使用免疫细胞化学和酶联免疫吸附试验评估涉及纤维化的细胞内促纤维化蛋白、细胞外基质蛋白和促炎细胞因子。

结果

与阳性对照 TGF-β1 组相比,LBP 预处理细胞的α-平滑肌肌动蛋白(成肌纤维细胞的标志物)、波形蛋白的表达显著降低(p<0.05),以及在瘢痕组织中发现的细胞外基质蛋白 II 型和 III 型胶原也显著降低(p<0.05)。此外,LBP 预处理细胞的促炎细胞因子白细胞介素 6 和白细胞介素 8 的分泌显著降低(p<0.05)。LBP 预处理和对照组之间,细胞包被水凝胶的收缩和硬度没有显著差异。LBP 预处理的成纤维细胞的血管生成因子表达和不受控制的增殖也受到抑制(p<0.05)。

结论

我们的结果表明,LBP 减少了体外角膜损伤中的促纤维化蛋白和促炎细胞因子。我们认为,LBP 作为一种天然中药,可能是一种新型的角膜屈光手术前的局部预处理选择,具有改善的预后。

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