| 외안F-035 |
| GSK3β 경로를 통한 ROCK 억제제 Y-27632의 각막윤부줄기세포의 증식과 분화 조절 |
| 한림대학교 의과대학 안과학교실(1), 서울대학교 의과대학 안과학교실(2), 서울대학교 의과대학 분당서울대병원 안과학교실(3), 성균관대학교 의과대학 안과학교실(4) |
| 이영복(1), 현준영(2,3), 정의상(4), 정태영(4), 이가영(1), 위원량(2), 신영주(1) |
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목적 : To investigate the effect of Rho-associated coiled kinase (ROCK) inhibitor Y-27632 on cultured corneal limbal stem cells.
방법 : Human limbal stem cells obtained from discarded corneal-scleral rings after penetrating keratoplasty were cultured. The cells were treated with EGF or a ROCK inhibitor or a combination of EGF and ROCK inhibitor. Periodic acid-Schiff (PAS) staining and immunofluorescence staining of p63, ABCG2, cytokeratin-7 (CK-7), cytokeratin-3/12 (CK-3/12) and MUC5AC were performed. The cell proliferation rate was measured using cell counting kit-8 assay. MUC5AC levels in the medium were measured by ELISA. CK-7, GSK3β, phosphrylated GSK3 β, and β-catenin expression was evaluated by western blotting.
결과 : The limbal stem cells were differentiated into both corneal epithelial cells and goblet cells. Goblet cells treated with the ROCK inhibitor and a combination of EGF and ROCK inhibitor showed the cytoplasmic granules by immunofluorescence staining. The cell proliferation rate increased with the ROCK inhibitor. Secreted MUC5AC levels in cells treated with a combination of EGF and the ROCK inhibitor were highest(p<0.05). GSK3β expressions in cells treated with the ROCK inhibitor and a combination of EGF and the ROCK inhibitor were lower compared to cells treated with control or EGF only.
결론 : Limbal stem cells were differentiated into both corneal epithelial cells and goblet cells. The ROCK inhibitor enhanced proliferation of corneal limbal stem cells and maturation and mucin secretion of goblet cells through the GSK3β pathway.
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