PAI-1反义寡核苷酸对肾间质细胞抑制作用的实验研究  

Inhibitory Effect of Plasminogen Activator Inhibitor-1 Antisense Oligonucleotides on NEK293 Cell Lines

作　　者：崔飞伦[1] 邱镇 陆洪兵[1] 姚震[1] 林大春[1] 

机构地区：[1]镇江市第一人民医院泌尿外科,江苏镇江212000 [2]镇江359医院泌尿外科,江苏镇江212000

出　　处：《解剖科学进展》2008年第1期70-72,共3页Progress of Anatomical Sciences

摘　　要：目的研究纤维蛋白溶解酶原激活物抑制因子-1(PAI-1)反义寡核苷酸(antisense oligonucleotides,ASODN)对大鼠肾间质细胞(NEK293)的抑制作用。方法采用RT-PCR、Western blot检测脂质体介导的PAI-1ASODN目的基因及蛋白的表达;通过细胞生长试验、流式细胞仪检测PAI-1 ASODN转染前后NEK293细胞增殖和凋亡的变化。结果PAI-1ASODN转染后可见NEK293中PAI-1mRNA及蛋白表达明显下调,与对照组相比较,可明显抑制细胞的增殖。诱导其凋亡(P<0.05)。结论PAI-1反义寡核苷酸具有明显抑制肾间质细胞增殖和诱导其凋亡的作用,转染脂质体介导的PAI-1 ASODN有望成为治疗肾纤维化的有效方法。Objective To explore the inhibitory effect of plasminogen activator inhibitor-1 （PAI-1） antisense oligonueleotides （ ASODN ） on the proliferation and apoptosis of NEK293 cell lines. Methods PAI-1 ASODN conjugated with isothioeyananate fluorescence was encapsulated by lipofeetamine 2000, and then added into the NEK293 cell culture media. The intraeellnlar distribution of PAI-1 ASODN was observed by fluorescence microscopy. The proliferation of NEK293 cell was measured by MTY test. The apoptosis of NEK293 ceil was measured by flow eytometer. The expression of PAI-1 was detected by RT-PCR and Western blot respectively. Result After PAI-1 ASODN was transfeeted into NEK293 cells ,we found that the expressions of PAI-1 mRNA and protein in NEK293 cells were significantly decreased or negative. PAI-1 ASODN inhibited the growth and proliferation of NEK293 cells and promoted the apoptosis of the cells compared with the control group （ P 〈0.01 ） .Conclusion The inhibited proliferation and induced apoptosis of NEK293 cells by PAI-1 ASODN suggest that PAI-1 ASODN with lipofectamine might be applieabal in the treatment of renal fibrosis.

关 键 词：纤维蛋白溶解酶原激活物抑制因子-1 反义寡核苷酸 肾间质细胞 增殖 凋亡 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]