Department of Pharmacology and Physiology, Faculty of Veterinary Medicine, University of Zaragoza, 50013 Zaragoza, Spain.
Exp Physiol. 2011 Nov;96(11):1151-62. doi: 10.1113/expphysiol.2011.060830. Epub 2011 Sep 2.
Alterations in intestinal motility are one of the features of sepsis induced by lipopolysaccharide (LPS). This study investigated the role of the nuclear transcription factor κB (NF-κB) in the LPS-induced duodenal contractility alterations, generation of reactive oxygen species (ROS) and production of cytokines in rabbit duodenum. Rabbits were treated with saline, LPS, sulfasalazine + LPS, pyrrolidinedithiocarbamate (PDTC) + LPS or RO 106-9920 + LPS. Contractility studies were performed in an organ bath. The formation of products of oxidative damage to proteins (carbonyls) and lipids (malondialdehyde and 4-hydroxyalkenals) was quantified in intestinal tissue and plasma. The protein expression of NF-κB was measured by Western blot. The DNA binding activity of NF-κB was evaluated by transcription factor activity assay. The expression of interleukin-1β, tumour necrosis factor α (TNF-α), interleukin-6, interleukin-10 and interleukin-8 mRNA was determined by RT-PCR. Sulfasalazine, PDTC and RO 106-9920 blocked the inhibitory effect of LPS on contractions induced by ACh in the longitudinal smooth muscle of rabbit duodenum. Sulfasalazine, PDTC and RO 106-9920 reduced the increased levels of malondialdehyde and 4-hydroxyalkenals and the carbonyls induced by LPS in plasma. Lipopolysaccharide induced the activation, translocation to the nucleus and DNA binding of NF-κB. Lipopolysaccharide increased the mRNA expression of interleukin-6 and TNF-α in duodenal tissue, and this effect was partly reversed by PDTC, sulfasalazine and RO 106-9920. In conclusion, NF-κB mediates duodenal contractility disturbances, the generation of ROS and the increase in the expression of interleukin-6 and TNF-α induced by LPS. Sulfasalazine, PDTC and RO 106-9920 may be therapeutic drugs to reduce these effects.
肠道运动的改变是脂多糖（LPS）诱导的败血症的特征之一。本研究探讨了核转录因子κB（NF-κB）在 LPS 诱导的十二指肠收缩改变、活性氧（ROS）生成和兔十二指肠细胞因子产生中的作用。兔子用生理盐水、LPS、柳氮磺胺吡啶+LPS、吡咯烷二硫代氨基甲酸盐（PDTC）+LPS 或 RO 106-9920+LPS 处理。在器官浴中进行收缩性研究。在肠组织和血浆中定量测定蛋白质（羰基）和脂质（丙二醛和 4-羟烯醛）氧化损伤产物的形成。通过 Western blot 测定 NF-κB 的蛋白表达。通过转录因子活性测定评估 NF-κB 的 DNA 结合活性。通过 RT-PCR 测定白细胞介素-1β、肿瘤坏死因子-α（TNF-α）、白细胞介素-6、白细胞介素-10 和白细胞介素-8 mRNA 的表达。柳氮磺胺吡啶、PDTC 和 RO 106-9920 阻断了 LPS 对兔十二指肠纵行平滑肌中 ACh 诱导收缩的抑制作用。柳氮磺胺吡啶、PDTC 和 RO 106-9920 降低了 LPS 诱导的血浆丙二醛和 4-羟烯醛以及羰基水平的升高。脂多糖诱导 NF-κB 的激活、核易位和 DNA 结合。脂多糖增加了十二指肠组织中白细胞介素-6 和 TNF-α 的 mRNA 表达，PDTC、柳氮磺胺吡啶和 RO 106-9920 部分逆转了这种作用。总之，NF-κB 介导 LPS 诱导的十二指肠收缩障碍、ROS 生成和白细胞介素-6 和 TNF-α 表达增加。柳氮磺胺吡啶、PDTC 和 RO 106-9920 可能是减少这些作用的治疗药物。
