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核因子-κB p65反义寡核苷酸对溃疡性结肠炎患者固有层单个核细胞中促炎细胞因子表达的影响

[The effects of nuclear factor-kappa B p65 antisense oligonucleotides on expression of proinflammatory cytokines in lamina propria mononuclear cells from patients with ulcerative colitis].

作者信息

Gan Huatian, Ouyang Qin, Chen Youqin, Liang Feng

机构信息

Department of Gastroenterology, West China Hospital, Sichuan University, Chengdu 610041.

出版信息

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2003 Jun;20(2):268-72.

Abstract

To investigate if nuclear factor-kappa B (NF-kappa B) p65 antisense oligonucleotides might affect the expression of NF-kappa B p65 and cytokines in lamina propria mononuclear cells(LPMC) from patients with ulcerative colitis (UC). LPMC were isolated from intestinal mucosal biopsy specimens from 3 patients with UC, and cultured with or without NF-kappa B p65 antisense oligonucleotides (5'-GGAACAGTTCGTCCTATGG-3'), missense oligonucleotides (5'-GGAACAGTTCGTCTATGG-3') and dexamethasone. NF-kappa B p65 expression was determined by western blot analysis. The expression of cytokine mRNA was studied by reversal transcription-polymerase chain reaction (RT-PCR). The cytokine levels were measured by enzyme linked immunosorbent assay. The results showed that NF-kappa B p65 antisense oligonucleotides resulted in down-regulation of NF-kappa B p65 expression, blocked the expression of IL-1 beta mRNA and IL-8 mRNA, and strikingly reduced the production of IL-1 beta and IL-8, and these effects were greater than those of dexamethasone in cultured LPMC from patients with UC(P < 0.05). Therefore, the application of NF-kappa B p65 antisense oligonucleotides may serve as a novel molecular approach for the treatment of patients with UC.

摘要

为研究核因子-κB(NF-κB)p65反义寡核苷酸是否会影响溃疡性结肠炎(UC)患者固有层单个核细胞(LPMC)中NF-κB p65及细胞因子的表达。从3例UC患者的肠道黏膜活检标本中分离出LPMC,并分别与NF-κB p65反义寡核苷酸(5'-GGAACAGTTCGTCCTATGG-3')、错义寡核苷酸(5'-GGAACAGTTCGTCTATGG-3')及地塞米松一起培养或不与它们一起培养。通过蛋白质印迹分析测定NF-κB p65的表达。采用逆转录-聚合酶链反应(RT-PCR)研究细胞因子mRNA的表达。通过酶联免疫吸附测定法测量细胞因子水平。结果显示,NF-κB p65反义寡核苷酸导致NF-κB p65表达下调,阻断白细胞介素-1β(IL-1β)mRNA和IL-8 mRNA的表达,并显著降低IL-1β和IL-8的产生,且在UC患者培养的LPMC中,这些作用大于地塞米松(P<0.05)。因此,应用NF-κB p65反义寡核苷酸可能成为治疗UC患者的一种新的分子方法。

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