Bai Ai-ping, Ouyang Qin, Gan Hua-tian, Wang Chun-hui
Department of Gastroenterology, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2004 Nov;35(6):778-80.
To investigate the effect of nuclear factor-kappa B activation on proinflammatory cytokine expression in intestinal epithelia when intestinal epithelia are in inflammatory condition.
Colonic adenocarcinoma HT29 cells were cultured in wells and TNF-alpha (10 ng/ml) was added into half of the wells. The supernatants were collected and measured for IL-8 after 3 hours, nuclear factor-kappaB (NF-kappaB) P65 and IL-8 mRNA were also examined by Western blotting and RT-PCR respectively.
It was found that in the condition of no stimuli, there is little IL-8 mRNA expression of HT29 cells, P65 is seldom in HT29 cells' nuclei, and the concentration of IL-8 in supernatant is just 172 ng/L. However, after being stimulated by TNF-alpha, a lot of P65 expression in nuclei and a large number of IL-8 mRNA expression of HT29 cells are shown and the concentration of IL-8 in supernatant (639 ng/L) is significantly higher than that of control (P<0.01).
The activation of nuclear factor-kappa B modulates the expression of some proinflammatory cytokine in intestinal epithelia, and regulates epithelia function in inflammatory reaction.
研究核因子-κB激活对处于炎症状态的肠上皮细胞促炎细胞因子表达的影响。
将结肠腺癌HT29细胞培养于孔板中,一半孔板中加入肿瘤坏死因子-α(TNF-α,10 ng/ml)。3小时后收集上清液检测白细胞介素-8(IL-8),同时分别采用蛋白质免疫印迹法和逆转录-聚合酶链反应(RT-PCR)检测核因子-κB(NF-κB)P65和IL-8 mRNA。
发现无刺激条件下,HT29细胞IL-8 mRNA表达很少,P65很少存在于HT29细胞核中,上清液中IL-8浓度仅为172 ng/L。然而,经TNF-α刺激后,HT29细胞核中出现大量P65表达和大量IL-8 mRNA表达,上清液中IL-8浓度(639 ng/L)显著高于对照组(P<0.01)。
核因子-κB的激活调节肠上皮细胞中某些促炎细胞因子的表达,并在炎症反应中调节上皮细胞功能。