p21^WAF1/CIP1在兔增生性玻璃体视网膜病变视网膜中的表达

The Expression of p21^WAF1/CIP1in Rabbit Proliferative Vitreoretinopathy

摘要: 目的：观察p21^WAF1/CIP1在兔增生性玻璃体视网膜病变(PVR)模型中的变化，探讨其在PVR发生发展的可能作用机制。方法：选取青紫蓝兔31只，实验组28只，正常对照组3只，组织病理学检查PVR病变情况，蛋白免疫印迹(Western blot)及逆转录聚合酶链反应(RT-PCR)检测不同时间点视网膜p21^WAF1/CIP1、CDK₂、cyclin E表达情况。结果：实验组7 d组织学检查示视网膜表面炎性细胞聚集，14 d时形成典型局部增殖条索，28 d时形成固定皱褶。Western blot结果显示p21^WAF1/CIP1相对表达量14 d时最低；CDK₂蛋白及cyclin E蛋白的相对表达量14 d最高，与其他时间点有统计学差异(F = 20.55, P = 0.00; F = 26.903, P = 0.000; F = 10.00, P = 0.00)。RT-PCR结果显示各组p21^WAF1/CIP1mRNA的相对表达量14 d时最低；CDK₂及cyclin E mRNA的相对表达量14 d最高，与其他时间点差异显著(F = 18.06, P = 0.00; F = 14.90, P = 0.00; F = 87.64, P = 0.00)。结论：p21^WAF1/CIP1在PVR表达下降，使PVR病情加重，这一作用可能是通过调节CDK₂和cyclin E的表达实现。

Abstract: Objective: To observe the expression of p21^WAF1/CIP1 during the course of experimental proliferative vitreoretinopathy (PVR) in rabbits and analyze its possible mechanism. Method: Thirty- one pigmented rabbits were divided into two groups, experimental group in 28 rabbits and normal control group in 3 rabbits. Histology was used to observe PVR progress. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) methods were used to detect the expression of p21^WAF1/CIP1, CDK₂, cyclin E protein or mRNA in retina on different set points. Results: Inflammatory cells infiltration was observed on day 7 in experimental rabbits’ retina. The proliferative typical strip of local retina was formatted on day 14 and fixed fold was noticed on the retina on day 28. Western blot method showed that the relative expression of p21^WAF1/CIP1 protein was lowest on day 14, and the relative expression of CDK₂ and cyclin E protein were highest on day 14. The difference with that in other set points had strong statistic significance (F = 20.55, P = 0.00; F = 26.903, P = 0.000; F = 10.00, P = 0.00). RT-PCR method also showed that the lowest relative expression on the relative expression of p21^WAF1/CIP1 mRNA was on day 14, and the highest relative expression on the relative expression of CDK₂ and cyclin E mRNA were noticed on day 14. The difference had strong statistic significance with that in other groups (F = 18.06, P = 0.00; F = 14.90, P = 0.00; F = 87.64, P = 0.00). Conclusions: The expression of p21^WAF1/CIP1 decreased during the course of PVR, which might contribute to the progress of PVR by regulation CDK₂ and cyclin E levels.