Basic approaches for the development of immunomodulatory and anti-inflammatory drugs acting on the signaling molecules

许多作用于IL-1和TNFa介导的信号转导的信号分子已经被发现。我们研究了粘着斑激酶(FAK)和肿瘤坏死因子受体相关因子(TRAF)在细胞因子信号转导和多种细胞凋亡诱导剂诱导的细胞凋亡中的作用。该系统被用于开发多种检测系统，以寻找新的免疫调节和抗炎药物。我们的研究结果如下：1)我们发现FAK-/-小鼠来源的FAK-/-MEF对TNFa和放线菌素D诱导的细胞凋亡比FAK/-细胞更敏感，这表明FAK-/-小鼠MEF对细胞死亡诱导剂具有抗凋亡作用。在这项研究中，FAK与RIP相关，而RIP在FAK-/-细胞的盘中被招募(Takahashi等人。此外，TRAF6-/-MEF对肿瘤坏死因子和放线菌素-D诱导的细胞死亡更为敏感。在这个系统中，活性氧物种(ROS)参与了细胞的死亡，因此，抗氧化剂BHA恢复了…更多的ROS诱导细胞死亡。因此，TRAF6参与了ROS诱导的细胞凋亡(Ichikawa et al.3)我们发现FAK过表达的HL-60细胞对包括TRAIL在内的多种细胞诱导剂诱导的细胞凋亡具有抵抗作用。相反，FAK过表达的HL-60对ATRA诱导的分化有抑制作用。这种重构条件是由于Rb蛋白的过度磷酸化和随后的c/EBPA失活(Hashimoto等人)。4)甘草甜素(GL)及其相关化合物可调节人肺成纤维细胞产生抗炎趋化因子、IL-8和嗜酸性粒细胞趋化因子-1。特别是，11-脱氧-GL和杂-30-羟基-GL能有效地抑制IL-8和嗜酸性粒细胞趋化因子-1(Matsui et al.2004、2006)。这些衍生物现在被设计用于体内试验。5)不同的蛋白酶体抑制剂被发现在人肺成纤维细胞中由肿瘤坏死因子和白介素4诱导嗜酸性粒细胞趋化因子-1和嗜酸性粒细胞趋化因子-3的作用不同(Rokudai等人)。这项研究适用于筛选作用于各种信号分子的免疫调节和抗炎药物。较少

Many signaling molecules have been identified which act on the IL-1 and TNFa-mediated signaling. We have studied the role of the focal adhesion kinase (FAK) and TNF receptor-associated factors (TRAF) in the cytokine signaling and apoptosis induced by various apoptosis inducing agents. This system was adopted to develop several assay systems to find new immunomodulatory and anti-inflammatory drugs. Our findings are followings.1) We found that FAK deficient MEF derived from FAK-/-mice is more susceptible to the TNFa and actinomycin D-induced apoptosis than the FAK+/-cells, indicating the anti-apoptotic role against the cell-death inducing reagents. In this study, FAK was associated with RIP, while RIP was recruited in the DISC in the FAK-/-cells (Takahashi et al. 2007).2) In addition, TRAF6-/-MEF is more sensitive to the TNFa and actinomycin-D induced cell death. In this system, reactive oxygen species (ROS) was involved in the cell-death and therefore, antioxidant reagent, BHA restored … More ROS-induced cell death. Thus, TRAF6 was involved in the ROS induced apoptosis (Ichikawa et al. 2006).Above two cell-culture systems are found to be easily applicable to the screening antioxidant reagents which protects from apoptosis-inducing conditions.3) We found that FAK-overexpressed HL-60 is resistant against apoptosis induced by the various apoptosis-inducing agents including TRAIL. In contrast, FAK-overexpressed HL-60 is reftractory to the ATRA-induced differentiation. This refactoctory condition was resulted from the hyperphosphorylation of Rb protein and subsequent the c/EBPa inactivation (Hashimoto et al. 2006).4) Glycyrrhizin (GL) and related compounds were found to regulate production of anti-inflammatory chemokines, IL-8 and eotaxin-1 in the human lung fibroblasts. In particularly, 11-deoxo-GL and Hetero-30-OH-GL were effective to inhibit IL-8 and eotaxin-1 (Matsui et al. 2004, 2006). These derivatives are now designed to be studied in in vivo assay.5) Various proteasome inhibitors were found to act differentially in the induction of eotaxin-1 and eotaxin-3 by the TNF and IL-4 in the human lung fibroblast (Rokudai et al. 2006).These study was applicable to the screening of the immunomodulatory and anti-inflammatory agents acting on the various signaling molecules. Less