Development of a novel therapy for severe acute pancreatitis by a specific inhibiiton of NF-κB- -Basic analysis using recombinant NLS-I k Bαadenoviral vector-

1999年，我们成功地将IκBα cDNA与核定位信号结合，构建了重组NLS-I κ Bα腺病毒载体。然而，IκBα和SV 40大T抗原的cDNA很难获得精确的核苷酸序列。我们还研究了cDNA构建体是否可以整合到腺病毒载体中，但也难以获得满意的结果。因此，我们在体内研究了NF-κB抑制剂如PDTC或NAC是否能减轻致死性胰腺炎（TCA胰腺炎）和非致死性水肿性胰腺炎（Cn胰腺炎）大鼠肺损伤模型中的肺损伤。PDTC或NAC预处理均能明显改善肺损伤，提高大鼠存活率。认为这种作用至少是通过阻断NF-κB活化抑制肺中TNA-α和IL-1 β的产生而产生的。2000年和2001年，我们继续生产重组NLS-I-κBα腺病毒载体。此外，我们还检测并证实了PAAF中的某些因子可以直接刺激血管内皮细胞或单核细胞，并在体外上调这些细胞中粘附分子和各种细胞因子的表达。我们还证实了抑制NF-κB可以阻断这些促炎分子的表达。

In 1999, we have to decelop a recombinant NLS-IκBα adenoviral vector in which IκBα cDNA was bound to a nuclear localizing signal. However, it was difficult to produce the cDNAs of IκBα and SV40 large T antigen with precise nudiotide sequences. We have also examined whether cDNA construct could be incorporated into the adenoviral vector or not, but it was also difficult to obtain satisfatory results. Therefore, we have studied in vivo whether the NF-κB inhibitors such as PDTC) or NAC could alleviate the lung injury in a model of lethal pancreatitis (TCA pancreatitis) and in a model with lung injury which was induced by a trasfer of the supematant of TCA pancreatitis ascites (P-AAF) into the rats with nonlethal edematous pancreatitis (Cn pancreatitis). In both models pretreatment with PDTC or NAC improved significantly the lung injury, and the survival of these rats was improved. The effects were considered to be brought about at least by the inhibition of TNA-α and lL-1 β production in the lung via a blockade of NF-κB activation. In 2000 and 2OO1, we have continued to produce the recombinant NLS-I-κBα adenoviral vector. Additionally, we have examined and verified the preset of some factors in the PAAF that could stimulate directly vascular endothelial cells or mono cytes and upregulate the expression of adhesion molecules and various cytoldnes in vitro in these cells. We have also confirmed that an inhibition of NF-κB could block the expression of these proinflammatory molecules.