Matrix metalloproteinase-9 is upregulated in inflammatory bowel disease. Barbiturate nitrate hybrid compounds have been designed to inhibit MMP secretion and enzyme activity. In this study, we investigated the mechanism of action of barbiturate-nitrate hybrid compounds and their component parts using models of intestinal inflammation in vitro. Cytokine-stimulated Caco-2 cells were used in all in vitro experiments. The NO donors SNAP and DETA-NONOate were used to study the effect of NO on MMP-9 mRNA. Mechanistic elucidation was carried out using the soluble guanylate cyclase (sGC) inhibitor, ODQ, and the cGMP analogue, 8-Bromo-cGMP. Further experiments were carried out to elucidate the role of NF-κB. NO donors exerted an inhibitory effect on MMP-9 mRNA in cytokine-stimulated cells. While the non-nitrate barbiturates had a limited effect on MMP-9 expression, the hybrid compounds inhibited MMP-9 expression through its NO-mimetic properties. No effect could be observed on mRNA for MMP-1 or MMP-2. The sGC inhibitior, ODQ, abolished the nitrate-barbiturate inhibition of MMP-9 gene expression, an effect which was reversed by 8-Br-cGMP. This study shows that the barbiturate scaffold is suitable for hybrid design as an MMP-9 inhibitor in cytokine-stimulated Caco-2 cells. The inhibition of MMP-9 levels was largely mediated through a reduction in its mRNA by a sGC/cGMP pathway mediated mechanism.

中文翻译：

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新型巴比妥酸盐-硝酸盐化合物通过cGMP介导的途径抑制肠道炎症中基质金属蛋白酶9基因表达的上调。

基质金属蛋白酶9在炎症性肠病中上调。已经设计了硝酸巴比妥酸盐杂化化合物来抑制MMP分泌和酶活性。在这项研究中，我们使用体外肠道炎症模型研究了巴比妥酸盐-硝酸盐杂化化合物及其组成部分的作用机理。细胞因子刺激的Caco-2细胞用于所有体外实验。用NO供体SNAP和DETA-NONOate研究NO对MMP-9 mRNA的影响。使用可溶性鸟苷酸环化酶（sGC）抑制剂ODQ和cGMP类似物8-Bromo-cGMP进行机理分析。进行了进一步的实验以阐明NF-κB的作用。没有供体对细胞因子刺激的细胞中的MMP-9 mRNA产生抑制作用。尽管非硝酸盐巴比妥酸盐对MMP-9的表达影响有限，但杂合化合物通过其NO模拟特性抑制MMP-9的表达。没有观察到对MMP-1或MMP-2的mRNA有影响。sGC抑制剂ODQ取消了硝酸盐-巴比妥酸盐对MMP-9基因表达的抑制作用，这种作用被8-Br-cGMP逆转。这项研究表明，巴比妥酸盐支架适合作为杂合设计，作为细胞因子刺激的Caco-2细胞中的MMP-9抑制剂。MMP-9水平的抑制主要是通过sGC / cGMP途径介导的机制降低其mRNA来介导的。这种作用被8-Br-cGMP所逆转。这项研究表明，巴比妥酸盐支架适合作为杂合设计，作为细胞因子刺激的Caco-2细胞中的MMP-9抑制剂。MMP-9水平的抑制主要是通过sGC / cGMP途径介导的机制降低其mRNA来介导的。这种作用被8-Br-cGMP所逆转。这项研究表明，巴比妥酸盐支架适合作为杂合设计，作为细胞因子刺激的Caco-2细胞中的MMP-9抑制剂。MMP-9水平的抑制主要是通过sGC / cGMP途径介导的机制降低其mRNA来介导的。