Choi Chang Hwan, Kim Tae Il, Lee Sang Kil, Yang Kyung Min, Kim Won Ho
Department of Internal Medicine, Institute of Gastroenterology, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul, South Korea.
Scand J Gastroenterol. 2008 Aug;43(8):938-47. doi: 10.1080/00365520801965373.
To evaluate the anti-inflammatory effect of Lactobacillus casei rhamnosus GG (LGG) and its conditioned media (CM) in stimulated Caco-2 cells and to characterize the components of LGG that have the anti-inflammatory effect.
Caco-2 cells were stimulated with IL-1beta with or without LGG or LGG-CM. Production of IL-8 was measured by enzyme-linked immunosorbent assay (ELISA). The transcriptional activities of the IL-8 gene and the NF-kappaB-responsive gene were evaluated by a transient transfection of the luciferase reporter gene. The effect on IkappaBalpha degradation was evaluated by Western blot analysis. To determine the nature of the immunomodulatory molecules, the LGG was modified to the following: treated with antibiotics, 4% formaldehyde, incubation at 95 degrees C, or sonication.
We demonstrated that the pretreatment of Caco-2 cells with LGG significantly inhibited IL-1beta-induced IL-8 production. Furthermore, LGG attenuated the IL-1beta-induced transcriptional activation of the IL-8 gene and the NF-kappaB-responsive gene, and attenuated the IL-1beta-induced IkappaBalpha degradation. Formaldehyde-fixed or antibiotics-treated LGG maintained the inhibitory effect, but heated LGG lost this effect. Sonicated LGG debris had a similar inhibitory effect with whole bacterial cells. LGG-CM attenuated IL-1beta-induced IL-8 production. This effect was maintained even when the conditioned media were heated.
LGG inhibited IL-1beta-induced IL-8 production in Caco-2 and this effect occurred at the transcriptional level, at least in part, by inhibition of the NF-kappaB signaling pathway. Both the structural material of LGG and the soluble factor secreted from LGG inhibited the IL-1beta-induced IL-8 production, and thus different substances may cause the effects.
评估鼠李糖乳杆菌GG（LGG）及其条件培养基（CM）对经刺激的Caco-2细胞的抗炎作用，并鉴定LGG中具有抗炎作用的成分。
用白细胞介素-1β（IL-1β）刺激Caco-2细胞，同时加入或不加入LGG或LGG-CM。通过酶联免疫吸附测定（ELISA）检测IL-8的产生。通过荧光素酶报告基因的瞬时转染评估IL-8基因和核因子κB（NF-κB）反应性基因的转录活性。通过蛋白质免疫印迹分析评估对IκBα降解的影响。为确定免疫调节分子的性质，将LGG进行如下处理：用抗生素处理、用4%甲醛处理、在95℃孵育或超声处理。
我们证明，用LGG预处理Caco-2细胞可显著抑制IL-1β诱导的IL-8产生。此外，LGG减弱了IL-1β诱导的IL-8基因和NF-κB反应性基因的转录激活，并减弱了IL-1β诱导的IκBα降解。甲醛固定或抗生素处理的LGG保持了抑制作用，但加热后的LGG失去了这种作用。超声处理的LGG碎片与完整细菌细胞具有相似的抑制作用。LGG-CM减弱了IL-1β诱导的IL-8产生。即使条件培养基被加热，这种作用仍能保持。
LGG抑制Caco-2细胞中IL-1β诱导的IL-8产生，且这种作用至少部分是通过抑制NF-κB信号通路在转录水平上发生的。LGG的结构物质和LGG分泌的可溶性因子均抑制IL-1β诱导的IL-8产生，因此可能是不同物质导致了这些作用。
