Department of Ophthalmology, First Affiliated Hospital of Nanchang University, Nanchang, P.R. China.
Department of Otolaryngology, First Affiliated Hospital of Nanchang University, Nanchang, P.R. China.
Invest Ophthalmol Vis Sci. 2020 Sep 1;61(11):19. doi: 10.1167/iovs.61.11.19.
This study investigated the role of S100 calcium binding protein A4 (S100A4) in corneal wound healing and the underlying mechanism of the S100A4-mediated PI3K/Akt/mammalian target of rapamycin (mTOR) pathway.
The rabbit corneal alkali burn model was established in vivo. S100A4 expression, wound healing, inflammation, and autophagy in rabbit cornea after alkali burn were detected. The NaOH-treated rabbit corneal stromal cells (rCSCs) were transfected with overexpressed S100A4 or silencing S100A4 to examine the effect of S100A4 on corneal wound healing in vitro. The effect of S100A4 on cell viability, proliferation, migration, invasion, fibrosis, and autophagy of rCSCs after alkali burn was analyzed. Then the functional rescue experiments were carried out. The PI3K inhibitor, LY294002, was used to elucidate the PI3K/Akt/mTOR signaling pathway in rCSCs.
S100A4 silencing promoted rabbit corneal wound healing by inhibiting fibrosis and inflammation and promoting autophagy in alkali-burned cornea, corresponding to increased levels of LC3, Beclin 1, and Atg4B but lowered α-smooth muscle actin, TNF-ɑ, and p62 levels. Moreover, silencing S100A4 inhibited proliferation, migration, invasion, and fibrosis of NaOH-treated rCSCs and promoted the differentiation of rCSCs into corneal cells and the autophagy of damaged rCSCs. The inhibitory role of S100A4 in wound healing was achieved via activation of the PI3K/Akt/mTOR pathway.
S100A4 silencing confers a promising effect on wound healing of alkali-burned cornea by blocking the PI3K/Akt/mTOR pathway, supporting the advancement of corneal gene therapies for wound healing.
本研究旨在探讨 S100 钙结合蛋白 A4（S100A4）在角膜伤口愈合中的作用及其介导的 PI3K/Akt/哺乳动物雷帕霉素靶蛋白（mTOR）通路的潜在机制。
建立体内兔角膜碱烧伤模型。检测兔角膜碱烧伤后 S100A4 表达、伤口愈合、炎症和自噬情况。用过表达 S100A4 或沉默 S100A4 转染 NaOH 处理的兔角膜基质细胞（rCSCs），观察 S100A4 对体外角膜伤口愈合的影响。分析 S100A4 对 rCSCs 碱烧伤后细胞活力、增殖、迁移、侵袭、纤维化和自噬的影响。然后进行功能挽救实验。用 PI3K 抑制剂 LY294002 阐明 rCSCs 中的 PI3K/Akt/mTOR 信号通路。
S100A4 沉默通过抑制纤维化和炎症、促进碱烧伤角膜自噬来促进兔角膜伤口愈合，相应地增加 LC3、Beclin 1 和 Atg4B 的水平，降低 α-平滑肌肌动蛋白、TNF-ɑ 和 p62 的水平。此外，沉默 S100A4 抑制 NaOH 处理的 rCSCs 的增殖、迁移、侵袭和纤维化，促进 rCSCs 向角膜细胞分化和损伤 rCSCs 的自噬。S100A4 通过激活 PI3K/Akt/mTOR 通路抑制伤口愈合。
沉默 S100A4 通过阻断 PI3K/Akt/mTOR 通路对碱烧伤角膜的伤口愈合有良好的效果，为伤口愈合的角膜基因治疗提供了支持。
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