论文部分内容阅读
目的探讨微型角膜刀法准分子激光角膜上皮瓣下磨镶术(Epi-LASIK)与乙醇浸润法准分子激光角膜上皮瓣下磨镶术(LASEK)对角膜基质细胞的影响研究。方法 50只新西兰大白兔随机编号,其中48只(96只眼)随机一只眼行 Epi-LASIK,另一只眼行 LASEK,余2只(4只眼)未进行手术,作为对照组。采用核苷酸末端转移酶介导的 dUTP 缺口标记(TUNEL)法观察角膜基质细胞的凋亡情况、Ki-67和α平滑肌肌动蛋白(α-SMA)的免疫组化和免疫印迹法测定角膜组织α-SMA 蛋白来观察角膜基质细胞的增殖与向肌成纤维母细胞转化情况,进行计数与半定量分析,对 haze 形成影响进行分析。结果 Epi-LASIK 与 LASEK 术后角膜基质中均有较多的 TUNEL 阳性细胞出现,在术后24h 达最高峰,LASEK 术后1周内细胞凋亡情况较 Epi-LASIK 明显(t=3.63、7.80、4.34、2.95,均P<0.01)。术后1周基质 Ki-67阳性细胞表达增高,72h 时达高峰,两组在1周内差异有统计学意义(t=3.81、5.85、5.09、5.59,均 P<0.01)。α-SMA 阳性细胞在术后1周明显出现,3个月时仍有较高表达,在术后1个月时阳性细胞数达高峰,α-SMA 蛋白表达量的情况与α-SMA 阳性细胞数变化一致,均提示 LASEK 术后增高的情况较 Epi-LASIK 差异有统计学意义(t=2.97、7.19、6.73,均 P<0.01)。结论 Epi-LASIK 较 LASEK 有着更少的角膜基质细胞凋亡、更弱的增殖以及向肌成纤维母细胞的转化,提示 Epi-LASIK 较 LASEK 有着更轻的 haze 反应几率。
Objective To investigate the effect of microkeratomes excimer laser keratomileusis (Epi-LASIK) and ethanol-infiltration excimer laser epithelial keratomileusis (LASEK) on corneal stromal cells. Methods Fifty New Zealand white rabbits were randomly assigned. Epi-LASIK was performed in 48 eyes (96 eyes) and LASEK in one eye. The other two eyes (4 eyes) were not operated as control group. The apoptosis of corneal stromal cells was observed by dUTP nick-end labeling (TUNEL), the expressions of Ki-67 and α-SMA were measured by immunohistochemistry and immunoblotting Α-SMA protein was observed to observe the proliferation of corneal stromal cells and the transformation into myofibroblasts, counting and semi-quantitative analysis of haze formation were analyzed. Results There were more TUNEL-positive cells in the corneal stroma after Epi-LASIK and LASEK, with the highest level at 24 hours after surgery. The apoptosis rate in EAC-LASIK and LASEK groups was higher than that in Epi-LASIK (t = 3.63, 7.80 , 4.34, 2.95, all P <0.01). The expression of Ki-67 positive cells increased at 1 week and peaked at 72 hours. There was significant difference between the two groups within 1 week (t = 3.81,5.85,5.09,5.59, all P <0.01). α-SMA positive cells appeared significantly at 1 week after operation and remained high at 3 months. The number of α-SMA positive cells reached the peak at 1 month after operation. The expression of α-SMA protein was positively correlated with α-SMA positive cells (P <0.01). All of them showed that the changes of postoperative LASEK were more significant than those of Epi-LASIK (t = 2.97, 7.19, 6.73, all P <0.01). Conclusion Epi-LASIK has less corneal stromal cell apoptosis, weaker proliferation and myofibroblast transformation compared with LASEK, suggesting that Epi-LASIK has a lighter haze reaction rate than LASEK.