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

许多信号分子作用于IL-1和tnfa介导的信号传导。我们研究了局灶黏附激酶（FAK）和TNF受体相关因子（TRAF）在细胞因子信号转导和各种细胞凋亡诱导剂诱导的细胞凋亡中的作用。该系统被用于开发几种检测系统，以发现新的免疫调节和抗炎药物。我们的发现如下。1)我们发现FAK-/-小鼠的FAK缺陷MEF比FAK+/-细胞更容易发生TNFa和放线菌素d诱导的细胞凋亡，表明FAK+/-细胞对细胞死亡诱导试剂具有抗凋亡作用。在本研究中，FAK与RIP相关，而RIP在FAK-/-细胞的DISC中被招募（Takahashi et al. 2007）。2)此外，TRAF6-/- mef对TNFa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（ROS）参与了细胞的死亡，因此抗氧化剂BHA恢复了ROS诱导的细胞死亡。因此，TRAF6参与了ROS诱导的细胞凋亡（Ichikawa et al. 2006）。发现上述两种细胞培养体系易于适用于筛选抗氧化试剂，以防止细胞凋亡诱导条件。3)我们发现fak过表达的HL-60对包括TRAIL在内的多种凋亡诱导剂诱导的细胞凋亡具有抗性。相反，fak过表达的HL-60对atra诱导的分化具有抗性。这种情况是由于Rb蛋白的过度磷酸化和随后的c/EBPa失活造成的（Hashimoto et al. 2006）。4)发现甘草酸（Glycyrrhizin， GL）及其相关化合物可调节人肺成纤维细胞中抗炎趋化因子、IL-8和eotaxin-1的产生。特别是，11-deoxo-GL和Hetero-30-OH-GL能有效抑制IL-8和eotaxin-1 （Matsui et al. 2004,2006）。这些衍生物现在被设计用于体内试验研究。5)在人肺成纤维细胞中，各种蛋白酶体抑制剂在TNF和IL-4诱导eotaxin-1和eotaxin-3的作用中存在差异（Rokudai et al. 2006）。本研究可用于筛选作用于各种信号分子的免疫调节剂和抗炎剂。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