p27^(Kip1)高表达对兔眼滤过术后成纤维细胞增殖作用的影响  

Effect of p27^(Kip1) overexpression on fibroblasts proliferation in rabbit after filtration surgery

作　　者：杨建刚[1] 何娜[2] 孙鹏锐 王小娣[3] 姚囯敏 刘倩影[2] 

机构地区：[1]西安市第一医院眼科,陕西省西安市710002 [2]西安交通大学医学院第二附属医院眼科,陕西省西安市710004 [3]西安医学院附属医院眼科,陕西省西安市710077

出　　处：《眼科新进展》2015年第2期120-124,共5页Recent Advances in Ophthalmology

基　　金：陕西省教育厅科研计划项目(编号:11JK705);西安市科技计划项目[编号:HM1116(5)]~~

摘　　要：目的研究外源性p27Kip1对兔眼滤过术后成纤维细胞p27Kip1、PCNA和AgN OR表达的影响。方法构建腺病毒为载体的p27Kip1(Ad-p27),长耳白兔24只随机分为Adp27组和对照组,行单眼滤过术,术中结膜下分别注射Ad-p27和磷酸盐缓冲液,术后7d、14 d、21 d和28 d免疫组织化学染色法评价手术区成纤维细胞p27Kip1和PCNA的表达,Ag NOR染色观察成纤维细胞核仁的Ag NOR颗粒计数。结果 Ad-p27作用后p27Kip1在Tenon囊成纤维细胞主要为核表达,术后7 d表达即增高,阳性细胞为79.17%,细胞评分为(6.40±0.67)分,术后28 d时阳性细胞为27.03%,评分为(4.97±0.83)分;对照组术后7 d和28 d时细胞评分分别为(4.88±0.79)分和(2.45±0.67)分,两组相比差异均具有统计学意义。Ad-p27组PCNA的表达降低,术后14 d始细胞评分与对照组相比差异具有统计学意义,持续至氉术后28 d。术后28 d内Ad-p27组Ag NOR染色始终为低表达,术后7 d颗粒计数为(2.18±0.72)个,较对照组的(4.04±0.68)个显著降低(P=0.000),28 d时颗粒计数为(2.13±0.69)个,较对照组的(4.06±0.85)个显著降低(P=0.000)。结论 p27Kipl高表达可以抑制兔眼滤过术后Tenon囊成纤维细胞的增殖,减少术后瘢痕形成,提高手术成功率。Objective To investigate the effects of exogenous P27Kipl on P27Kipl , PCNA and AgNOR expression in Tenon＇ s fibroblasts on rabbit eyes after filtration sur- gery. Methods The P27Kipl -expressing adenovirus （Ad-p27） was constructed. Twenty- four rabbits were divided randomly into Ad-p27 group and control group. The unilateral eyes were performed filtration surgery. Ad-p27 or PBS was injected into subconjunctiva during surgery, respectively. The expressions of P27Kipl or PCNA in fibroblasts were ana- lyzed at surgical sites at 7 days,14 days,21 days and 28 days after surgery by immuno- histochemistry. The numbers of nucleolar AgNOR particles were counted in fibroblasts by AgNOR staining. Results P27Kipl expression was mainly observed in the nucleus of fibroblasts induced by Ad-p27. The expression was increased significantly at 7 days. The positive cells were 79.17% in Ad-p27 group,the cell scores were 6.40 ±0.67 ;The ex- pression remained high level at 28 days, the positive cells were 27.03% , and the cell scores were 4.97 ± 0.83 ; The cell scores in control group was 4.88 ± 0.79 and 2.45 ± 0. 67,there were statistical differences between two groups. The expression of PCNA was decreased significantly in Ad-p27 group, there was statistical difference in cell scores at 14 days compared with control group. The expressions of AgNOR were persist- ently decreased within 28 days in Ad-p27 group, the number of AgNOR particles at 7 days and 28 days were 2. 18 ± 0.72 and 2. 13± 0.69, respectively, compared with the control group （4.04 ± 0.68 and 4.06 ± 0. 85 ）, there were significant differences （ both P = 0.000）. Conclusion The overexpression of 1）27 rap1 can inhibit the proliferation of Tenon fibroblasts after filtration surgery,inhibit the scar formation and promote the sur- gical outcome.

关 键 词：细胞周期依赖性激酶抑制剂 P27KIP1 青光眼 滤过术 细胞增殖 兔 

分 类 号：R775[医药卫生—眼科]