NF-κB靶向性哑铃形“圈套”寡核苷酸抑制肾小球系膜细胞IL-6的表达  

Inhibitory effect of circular dumbbell decoy ODNs targeting NF-κB on IL-6 expression in mesangial cells

作　　者：张建国[1] 王惠民[1] 国华[2] 郝天智[2] 杨文军[2] 

机构地区：[1]第三军医大学附属大坪医院野战外科研究所肾内科,重庆400042 [2]第三军医大学附属大坪医院野战外科研究所第一研究室,重庆400042

出　　处：《第三军医大学学报》2003年第19期1768-1771,共4页Journal of Third Military Medical University

基　　金：重庆市科委应用基础项目 ( 41A11537)

摘　　要：目的　设计合成靶向NF κB的哑铃形“圈套”ODNs ,并检测其对NF κB转录活性和肾小球系膜细胞IL 6表达的抑制效应。方法　采用电泳迁移率改变实验 (EMSA)体外检测哑铃形圈套ODNs对NF κB转录活性的抑制效应。提取大鼠肾小球系膜细胞 ,随机分为正常对照组、LPS刺激组、哑铃形圈套ODNs处理组、无关圈套ODNs处理组和脂质体处理组。采用阳离子脂质体将 2、4、8mg L不同剂量哑铃形圈套ODNs转染大鼠肾小球系膜细胞 ,6h后用LPS刺激 ,收集转染后 8、12、18h上清和细胞。用酶联免疫吸附试验 (ELISA)法检测上清IL 6蛋白表达 ,逆转录PCR(RT PCR)法检测IL 6mR NA转录。结果　哑铃形圈套ODNs在体外能有效地抑制NF κB与其顺式元件的结合 ;4、8mg L剂量哑铃形圈套ODNs可明显抑制LPS诱导的大鼠肾小球系膜细胞IL 6转录和表达。结论　靶向NF κB的哑铃形圈套ODNs在体外可抑制NF κB的转录活性 ,从而对体外培养的肾小球系膜细胞炎性因子ILObjective To investigate the inhibitory effects of circular dumbbell decoy oligodeoxynucleotides(ODNs) targeting NF κB on the transcriptional activity of NF κB binding to its cis elements and the expression of IL 6 in mesangial cells. Methods The inhibitory effect of circular dumbbell decoy ODNs on the transcriptional activity of NF κB was detected by electrophoretic mobility shift assay (EMSA). Mesangial cells extracted from rats were randomly divided into normal control, LPS stimulated, circular dumbbell decoy ODNs treated, scrambled decoy ODNs treated and cationic liposome treated groups. Circular dumbbell decoy ODNs at the doses of 2 mg/L, 4 mg/L and 8 mg/L were transfected respectively into mesangial cells from rats using cationic liposome vector. Mesangial cells were stimulated with LPS at 6 h after circular dumbbell decoy ODNs transfection. Then, the transfected supernatant and cells were collected at 8, 12 and 18 h. IL 6 mRNA expression in mesangial cells was detected by RT PCR and IL 6 protein level by ELISA. Results Circular dumbbell decoy ODNs could effectively inhibit the DNA binding activity of NF кB to its cis elements in vitro . Circular dumbell decoy ODNs at the doses of 4 and 8 mg/L could effectively inhibit the expression levels of IL 6 mRNA and protein in mesangial cells. Conclusion Circular dumbbell decoy ODNs targeting NF κB could effectively inhibit the expression of IL 6 in mesangial cells through the inhibition of NF κB activity.

关 键 词：NF-ΚB 哑铃形圈套ODNs 白细胞介素6 

分 类 号：Q524[生物学—生物化学] R322.61[医药卫生—人体解剖和组织胚胎学]