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目的探讨bFGF在中期保存角膜过程中对角膜细胞凋亡的影响,以期减少中期保存角膜细胞凋亡,为改良角膜中期保存液提供实验依据。方法配制中期保存液保存新西兰大白兔角膜,设对照组为保存液保存,实验组为保存液加入bFGF(10ng/ml),两组在30～34℃恒温条件下保存16～18h后,转入4℃低温保存,在保存3、5、7d分别取保存角膜片行30～34℃2h复温,固定、切片、TUNEL原位标记免疫荧光和电镜检测保存角膜细胞凋亡。结果加入bFGF组保存角膜水肿较轻,皱褶较少,透明度较高,角膜细胞形态和结构较好;bFGF组保存角膜与对照组比较细胞凋亡较少,差异有显著性(p<0.05)。结论中期保存角膜细胞会发生不同程度的细胞凋亡,导致细胞活性丧失;bFGF在中期角膜保存中有抑制和减轻角膜细胞凋亡发生,保持角膜细胞结构和功能的作用;bFGF可以应用于角膜中期保存。
Objective To investigate the effect of bFGF on the apoptosis of corneal cells in the process of preserving the cornea in the medium term in order to reduce the apoptosis of corneal cells in the medium term and provide experimental evidence for improving the corneal mid-term preservation solution. Methods The cornea of New Zealand white rabbits were preserved in the mid-term preservation solution. The control group was preserved in preservation solution. The experimental group was bFGF (10 ng / ml). The two groups were kept at 30 ~ 34 ℃ for 16 ~ 18 h. The cells were stored at 4 ℃ for 3 and 7 days. The preserved corneal slices were re-incubated at 30-34 ℃ for 2 hours, then fixed and sectioned. The apoptotic cells were preserved by TUNEL in situ immunofluorescence and electron microscopy. Results The bFGF group had less corneal edema, less folds, higher transparency and better morphology and structure of corneal cells. Compared with the control group, the bFGF group showed less apoptotic cells (p <0.05) . CONCLUSIONS: There are different degrees of cell apoptosis in the medium-term preservation of corneocytes, resulting in the loss of cell activity. BFGF can inhibit and reduce the apoptosis of keratocytes and maintain the structure and function of corneal cells in mid-term corneal preservation. BFGF can be used in the mid-cornea save.