Agrin 通过 Hippo-Yap 信号通路促进角膜缘干细胞增殖和角膜伤口愈合。

PURPOSE

To investigate the effect and mechanism of Agrin on limbal stem cell proliferation and corneal wound healing.

METHODS

Limbal stem cells were isolated and treated with different concentrations of Agrin. CCK-8 and cell proliferation markers (Ki67 and pH3) were detected to evaluate cell numbers or proliferative potential of limbal stem cells. The corneal epithelium wound model was induced by debridement of central corneal epithelial, and the effects of Agrin on limbal stem cell proliferation and corneal epithelial wound healing rate were determined.

RESULTS

Agrin promoted the proliferation of cultured limbal stem cells in vitro and increased the expression level of p63α rather than keratin 12. Furthermore, Agrin accelerated the wound healing rate of corneal epithelium through activating limbal stem cell proliferation in vivo. In terms of mechanism, Agrin could facilitate the dephosphorylation of Yap1, which contributed to the nuclear translocation of Yap1 and expression of Cyclin D1, and subsequently promoted proliferation of limbal stem cells.

CONCLUSIONS

Agrin promotes the proliferation of limbal stem cells and accelerates the healing rate of corneal wound through Hippo-Yap signaling pathway.

目的

研究 Agrin 对角膜缘干细胞增殖和角膜伤口愈合的作用及机制。

方法

分离角膜缘干细胞并使用不同浓度的 Agrin 进行处理。通过 CCK-8 法和细胞增殖标志物（Ki67 和 pH3）检测来评估角膜缘干细胞的数量或增殖潜能。通过中央角膜上皮刮除术诱导角膜上皮创面模型，以确定 Agrin 对角膜缘干细胞增殖和角膜上皮创面愈合率的影响。

结果

Agrin 促进体外培养的角膜缘干细胞增殖，并增加 p63α而不是角蛋白 12 的表达水平。此外，Agrin 通过激活体内角膜缘干细胞增殖来加速角膜上皮的愈合速度。就机制而言，Agrin 可以促进 Yap1 的去磷酸化，这有助于 yap1 的核转位和 Cyclin D1 的表达，从而促进角膜缘干细胞的增殖。