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取正常小鼠、糖尿病2周(DM1组)及20周(DM2组)小鼠的视网膜样本,TUNEL法检测细胞凋亡及存在的部位;免疫印迹法检测一氧化氮合酶(iNOS、nNOS)的表达;竞争性ELISA法测定硝基酪氨酸含量,检测视网膜损伤程度,探讨一氧化氮合酶与早期糖尿病小鼠(2、20周)视网膜细胞凋亡的关系。结果表明:对照组未发现凋亡细胞,DM1组在神经节细胞层可见少量凋亡细胞,DM2组神经节细胞层、内核层与外核层均见大量凋亡阳性细胞;免疫印迹法检测发现,与对照组比较,iNOS在DM1和DM2组中表达显著增多,组间差异不显著;nNOS在DM1组表达减少,在DM2组中则几乎消失;视网膜硝基酪氨酸的含量DM2组显著高于对照组与DM1组。证明早期糖尿病视网膜的细胞凋亡与iNOS表达升高及nNOS表达降低有关。
Retinal samples of normal mice, diabetic DM2 and DM2 mice were detected by TUNEL method. Nitric oxide synthase (iNOS, nNOS) . The content of nitrotyrosine was measured by competitive ELISA and the degree of retinal damage was detected. The relationship between nitric oxide synthase and retinal cell apoptosis in early diabetic mice (2, 20 weeks) was also explored. The results showed that apoptotic cells were not found in the control group. A small amount of apoptotic cells were observed in the ganglion cell layer in the DM1 group. A large number of apoptotic positive cells were found in the ganglion cell layer, the inner nuclear layer and the outer nuclear layer in the DM2 group. Compared with control group, the expression of iNOS increased significantly in DM1 and DM2 group, but there was no significant difference between the two groups. The expression of nNOS decreased in DM1 group and almost disappeared in DM2 group. The content of nitrotyrosine in DM2 group was significantly higher In control group and DM1 group. Prove the early diabetic retinal apoptosis and iNOS expression increased and nNOS expression decreased.