NF-κB靶向性哑铃形圈套寡核苷酸抑制肾小球系膜细胞TGF-β_1的表达  

Inhibitory effects of circular dumbbell decoy ODNs targeting NF-κB on TGF-β_1 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肾内科,重庆400042 [2]第三军医大学野战外科研究所一室,重庆400042

出　　处：《第四军医大学学报》2004年第14期1266-1269,共4页Journal of the Fourth Military Medical University

基　　金：重庆市科委应用基础项目 (41A1 1 537)

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TGF β1 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 8,1 2 ,1 8h上清和细胞 .用酶联免疫吸附试验 (ELISA)法检测上清TGF β1 蛋白表达 ,逆转录PCR(RT PCR)法检测TGF β1 mR 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TGF β1 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TGF β1AIM: To study the inhibitory effects of circular dumbbell decoy oligodeoxynucleotides targeting NF κB on binding activity of NF κB with its cis elements and the expression of TGF β 1 in mesangial cells. METHODS: Circular dumbbell decoy ODNs competition inhibition effects were assayed by EMSA. Mesangial cells from rats were isolated and randomly divided into normal control, LPS stimulated, circular dumbbell decoy ODNs treated, scrambled decoy ODNs treated and cationic liposome treated groups. With carriers of cationic liposomes, 2, 4, and 8 mg/L decoy ODNs were respectively transfected into the cells and the mesangial cells were stimulated with LPS 6 h after circular dumbbell decoy ODNs transfection. Supernatants and cells were collected at 8, 12 and 18 h after decoy ODNs transfection. TGF β 1 mRNA expression in mesangial cells was assessed by RT PCR and TGF β 1 protein level was determined by ELISA. RESULTS: Circular dumbbell decoy ODNs effectively inhibited the DNA binding activity of NF кB in vitro. After using decoy ODNs, the expression levels of TGF β 1 mRNA and protein in mesangial cells significantly decreased. CONCLUSION: Circular dumbbell decoy ODNs targeting NF κB could effectively inhibit the expression of TGF β 1 in mesangial cells through specifically inhibiting the activity of NF κB.

关 键 词：NF-ΚB 哑铃形圈套ODNs 转化生长因子Β 

分 类 号：R322.61[医药卫生—人体解剖和组织胚胎学]