反义寡核苷酸封闭CD44基因表达对小梁细胞吞噬及聚集功能的影响  

Effect of CD44 suppression by antisense oligonucleotide on phagocytosis and aggregation of human trabecular meshwork cells

作　　者：李中国[1] 王林农[1] 张虹[2] 

机构地区：[1]南京医科大学附属南京第一医院眼科,江苏省南京市210006 [2]华中科技大学同济医学院附属同济医院眼科,湖北省武汉市430030

出　　处：《眼科新进展》2008年第11期801-804,共4页Recent Advances in Ophthalmology

基　　金：南京医科大学科技发展基金资助(编号:NY0566)~~

摘　　要：目的观察CD44反义寡核苷酸封闭CD44基因表达对人眼小梁细胞吞噬、聚集功能的影响,进一步探讨黏附分子CD44在原发性开角型青光眼(primary open angleglaucoma,POAG)发病过程中可能的作用。方法取意外死之后行角膜移植供体眼小梁细胞,实验分空白对照组,2.0μmol·L-1正义寡核苷酸,0.5μmol·L-1、1.0μmol·L-1、2.0μmol·L-1反义寡核苷酸共5组,采用硫代修饰的CD44反义寡核苷酸,通过脂质体介导转染体外培养的人眼小梁细胞。以乳胶微粒为标记定量观察CD44反义寡核苷酸封闭CD44基因表达对人眼小梁细胞吞噬功能的影响,计数法观察CD44反义寡核苷酸封闭CD44基因表达对人眼小梁细胞聚集功能的影响。结果3种浓度的CD44反义寡核苷酸处理的小梁细胞吞噬微粒数分别为58.2±18.8、47.5±16.9、34.9±16.3,明显低于对照组的75.9±13.4(P<0.05)及CD44正义寡核苷酸组的74.0±11.8(P<0.05),且呈浓度依赖性。而对照组与CD44正义寡核苷酸组间比较差异无统计学意义(P>0.05)。3种浓度的CD44反义寡核苷酸处理各组出现细胞聚集阳性平均次数分别为(24.50±5.26)次、(16.30±3.77)次、(9.25±1.71)次,明显低于对照组的平均(32.00±3.37)次(P<0.05)和CD44正义寡核苷酸组的(33.25±5.32)次(P<0.05),且呈浓度依赖性,而CD44正义寡核苷酸组与对照组比较差异无统计学意义(P>0.05)。结论CD44反义寡核苷酸封闭CD44基因表达后人眼小梁细胞吞噬功能降低以及小梁细胞的聚集功能受到抑制,黏附分子CD44可能通过影响小梁细胞的吞噬以及聚集功能参与了POAG发病过程。Objective To observe the effect of CD44 suppression by anti sense oligonucleotide on phagocytosis and aggregation of human trabecular meshwork cells, then to discuss the possible contribution of CD44 to the pathogenesis of primary open angle glaucoma （POAG）. Methods After accidental death, corneal transplanted trabecular meshwork cells were divided into control group, 2.0 μmol·L^-1 sense oligonucleotide group, 0.5 μmol·L^-1, 1.0 μmol·L^-1 and 2.0 μmol·L^-1 antisense oligoncleotide group, CD44-specific antisense oligonucleotide were delivered with cationic lipid to transfer human trabecular meshwork cells. Effect of CD44-specific antisense oligonucleotide on phagocytosis of trabecular meshwork cells was quantitively measured by using latex beads as markers; Cell count method was used to observe the effect of CD44-specific antisense oligonucleotide on cell aggregation. Results The number of trabecular meshwork cells inhibited by three different concentrations of CD44 antisense oligonucleotide was 58.2 ± 18.8,47.5 ± 16.9,34.9 ± 16.3 ,respectively and in a concentration-dependent manner, which were significantly lower than 75.9 ± 13.4 in control group（ P 〈 0.05 ） and than 74.0 ± 11. 8 in sense oligonucleotide （ P 〈 0.05 ）, while no statistical difference was found between control group and antisense oligonucleotide group （ P 〉 0.05 ）. The coverage positive cell aggregation times by three different antisense oligonueleotide was 24.50 ± 5.26,16.30 ± 3.77,9.25 ± 1.71, and in a concentration-dependent manner,which were significantly lower than 32.00 ± 3.37 of control group（ P 〈 0.05 ） and lower than 33.25 ± 5.32 of sense oligonucleotide group（ P 〈 0.05 ）, while no statistical difference was found between control group and sense oligonucleotide （ P 〉 0.05 ）. Conclusion CD44 antisense oligonncleotide can inhibit the phagocytosis and phagregation of human trabecular meshwork cells. CD44 may play a role in pathogenesis of POAG by affecting phagocytosis and aggregation

关 键 词：小梁细胞 CD44 反义寡核苷酸 吞噬 聚集 

分 类 号：R775.2[医药卫生—眼科]