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采用锥蓝 -茜素红联合染色和酶组织化学染色技术 ,对经过甘油干燥保存 1、3、6个月的猫角膜内皮细胞进行活性检测。结果显示保存不同时期的角膜内皮细胞经锥蓝 -茜素红联合染色后 ,细胞核均深度蓝染 ,蓝染率10 0 % ,细胞边界不清晰。经组织培养 7天后 ,细胞核蓝染率仍为 10 0 %。ACP、ATPase、SDH和苹果酸脱氢酶在各时期保存角膜内皮细胞中均可检测到。保存 1、3个月角膜中 ,四种酶与正常对比无明显差异 (P>0 .0 5 )。而保存6个月组 ,除 ATPase外 ,其余三种酶反应程度降低 (P<0 .0 5 )。干燥保存角膜培养 7天后 ,内皮细胞中均未再检测到酶活性 ,认为经甘油干燥保存的角膜内皮细胞不再具有活性。
Cone blue - alizarin red combined staining and enzymatic histochemical staining were used to detect the activity of endothelial cells of cat corneas dried for 1, 3, and 6 months after glycerol drying. The results showed that the corneal endothelial cells preserved at different stages were stained by cone blue - alizarin red and the nuclei were deeply blue-stained. The blue dye rate was 100% and the cell borders were not clear. After 7 days of tissue culture, the rate of nuclear blue dye was still 100%. ACP, ATPase, SDH and malate dehydrogenase were preserved in corneal endothelial cells at various times. There was no significant difference between the four kinds of enzymes and the normal ones in the preserved corneas for 1 and 3 months (P> 0.05). However, in the 6-month storage group, except for ATPase, the levels of the other three enzymes decreased (P <0.05). No enzymatic activity was detected in the endothelial cells after 7 days of dry preservation of the cornea, suggesting that corneal endothelial cells preserved in glycerol were no longer active.