糖尿病小鼠眼opticin蛋白表达的研究

Expression of opticin in the eyes of diabetic mice

目的 观察发生糖尿病与未发病非肥胖性糖尿病（NOD）小鼠玻璃体内opticin蛋白表达差异.方法 建立糖尿病NOD小鼠模型作为实验组.选用同龄NOD小鼠作为对照.颈椎脱臼法处死两组所有小鼠.摘除眼球,钝性分离玻璃体、视网膜、巩膜.采用蛋白质免疫印迹（Wesrern blot）和实时定量逆转录聚合酶链反应（RT-PCR）对比分析实验组和对照组玻璃体、视网膜、巩膜中opticin蛋白及其相关基因（OPTC）mRNA的表达.结果 Western blot检测显示,实验组和对照组玻璃体、视网膜及巩膜中均可见opticin蛋白表达条带,主要位于相对分子质量60×10~3处.实验组玻璃体、视网膜中opticin蛋白较对照组显著降低（t=4.42,4.58 P=0.002,0.002）,巩膜中opticin蛋白差异无统计学意义（t=0.27,P=0.794）.RT-PCR检测显示,OPTC mRNA表达分布规律为玻璃体、视网膜、巩膜依次递减.实验组玻璃体、视网膜中OPTC mRNA表达较对照组显著降低（t=3.30,2.48 P=0.01,0.04） 巩膜中OPTC mRNA表达差异无统计学意义（t=0.27,P=0.80）.结论 糖尿病NOD小鼠玻璃体、视网膜中opticin蛋白表达较未发病NOD小鼠受抑制.

Objective To observe the opticin expression in the eyes of non-obese diabetes （NOD） mice and non-diabetic NOD mice . Methods Twenty NOD mice were divided into diabetic group （experimental group） and non-diabetic group （control group）. All the mice were killed by cervical dislocation method. The eyes were harvested, and the vitreous, retina and sclera were separately collected. Western blot and real-time reverse transcription-polymerase chain reaction（RT-PCR）were respectively used to determine opticin protein and OPTC-mRNA levels. Results The opticin protein level in the vitreous and retina was lower in the experimental group（t = 4.42,4.58 P = 0. 002,0. 002）, but is same in the sclera between the 2 groups （t = 0. 27, P = 0. 794）. OPTC-mRNA level was vitreous〉 retina〉 selera. OPTC-mRNA levels of vitreous and retina in diabetic group were significantly Iower（t = 3.30,2. 48 P= 0. 01, 0.04） there was no statistical significant on OPTC mRNA of sclera between two groups（t = 0. 27, P = 0. 80）. Conclusion Expression of opticin was suppressed in retina and vitreous of diabetic mice.