核因子-κB“诱饵性”寡核苷酸抑制LPS诱导的肠上皮细胞TLR4、IL-8的表达

目的： 研究NF-κB decoy寡核苷酸对LPS诱导结肠癌细胞SW480后，对TLR4、IL-8表达的影响。方法：体外培养SW480细胞，用LPS(10 μg/L)刺激3 h后，用脂质体lipofectin 2000介导NF-κB decoy ODNs转染6 h，收集细胞上清液，用ELISA法检测IL-8；提取细胞mRNA，经 RT-PCR法检测TLR4 mRNA、IL-8 mRNA的表达。并设对照组、Scrambled ODNs组、lipofectin 2000组进行比较。结果：LPS刺激后TLR4 mRNA、IL-8 mRNA和IL-8的表达明显强于对照组；用NF-κB decoy寡核苷酸干预，可以明显抑制TLR4 mRNA、IL-8 mRNA和IL-8的表达。而Scrambled ODNs组和转染剂lipofectin 2000组对其没有明显影响。结论：NF-κB decoy ODNs有望成为治疗IBD的一种新型基因药品。

Inhibitory effect of NF - κB decoy ODN on expressions of TLR4 and IL -8 in LPS - induced intestinal epithelial cells

AIM: To study the effect of NF-κB decoy oligodeoxynucleotides(ODNs) on TLR4 and IL-8 expression in LPS-induced SW480 cells. METHODS: SW480 cells were cultured in vitro and stimulated for 3 h with LPS (10 μg/L). NF-κB decoy oligodeoxynucleotides mediated by lipofectin 2000 were added into the cell culture for 6 h. The supernatants were collected and messured for IL-8 by ELISA. TLR4 mRNA and IL-8 mRNA were examined by RT-PCR, respectively. The results were compared with control group, Scrambled ODNs group and lipofectin 2000 group. RESULTS: After SW480 cells were stimulated by LPS, TLR4 mRNA, IL-8 mRNA and IL-8 expressions were significantly increased, and the difference compared with control group was obvious. After treated with NF-κB decoy oligodeoxynucleotides, TLR4 mRNA, IL-8 mRNA and IL-8 expressions were significantly inhibited. The Scrambled ODNs group and lipofectin 2000 group had no effect on them. CONCLUSION: NF-κB decoy ODNs will become a new gene drug for treating inflammatory bowel disease(IBD).