辐射后兔晶体上皮细胞酸性磷酸酯酶的定量分析

Quantitative Analysis of ACP Activity in Rabbit lens Epithelial cells After Radiation

目的 研究电离辐射对离体培养的兔晶体上皮细胞中酸性磷酸酯酶的影响。方法 用6 0Coγ线单次照射培养的第三代兔晶体上皮细胞 ,经ACP细胞化学方法染色后 ,用图像分析技术对其进行定量分析。结果 1、晶体上皮细胞于培养后 3天可见有单个细胞从晶体上皮长出 ,至培养第三代的细胞经 30Gy6 0 Coγ线照射后 ,细胞生长速度变慢 ,细胞形态不规则。照射后ACP呈强阳性表达 ,照后 7天更为明显。电镜观察可见 ,晶体上皮细胞核仁肿胀 ,染色质稀少 ,内质网扩张 ,线粒体萎缩。溶酶体膜破裂 ,随着照后时间延长 ,病变更为明显。 2、定量结果显示 :各剂量组在照后3和 7天 ,ACP的面积、平均光密度、积分光密度均显著高于对照组 (P <0 .0 0 1) ,其中 7天各指标升高更为明显 ,此结果与电镜所见相一致。结果提示 ,电离辐射对细胞溶酶体膜造成损伤是ACP活力改变可能的机理 ,而后者可能是导致晶体上皮损伤和死亡的原因之一。

Objective To investigate the Effects of ion-radiation on ACP activity in cultured rabbit lens epithelial cells(LEC).Method After 60 Coγ ray single radiation, the cultured rabbit LEC was stained by ACP cytochemistry method, then analyzed by image analyzer.Results 1. After 3 days culture, it can be seen that single LEC growing from lens epithelia. After 30Gy 60 Coγ ray radiation, the cultured cells grew slowly and their morpohology became irregular. Strong positive expression of ACP activity was found on day 7 after irradiation, chromatin scarce, ER dilation, mitochondria atrophy, The pathologic changes become more obvions, with the prolongation after irradiation at 3 and 7 days after radiation…2.Quantitative analysis of ACP showed, the ACP area, MOD and IOD in all radiation groups were higher significantly than those control groups (p<0.001) especially, onday 7 after irradiation, which is in accord with the observation by EM.The results suggest that ion-radiation. could destroy the lysosomal membrane. of LEC and induce the increase ACP activity, while the later could be one of the reasons which cause LEC epithelia injury and death.