Gan Huatian, Ouyang Qin, Chen Youqin, Liang Feng
Department of Gastroenterology, West China Hospital, Sichuan University, Chengdu 610041.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2003 Jun;20(2):268-72.
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.
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。