Acute pancreatitis (AP) can lead to death; however, there is no specific treatment for AP. Screening of drugs for AP treatment is rarely performed. Compounds were screened in a primary pancreatic acinar cell and peritoneal macrophage coculture system. Compounds were used in vitro and in vivo. Compound targets were predicted and validated. Among the 18 nitrogen-containing heterocycles, Z10 was shown to decrease the cerulein plus lipopolysaccharide (CL)–induced secretion of both acinar digestive enzymes and macrophage cytokines. Z10 was also shown to ameliorate CL-induced or sodium taurocholate–induced AP in mice. Proteomics analysis and enzyme linked immunosorbent assay (ELISA) revealed that Z10 decreased the levels of D-dopachrome tautomerase (Ddt) within macrophages and those in the extracellular milieu under CL treatment. Z10 also decreased Ddt expression in AP mice. Moreover, exogenous Ddt induced cytokine and digestive enzyme secretion, which could be inhibited by Z10. Ddt knockdown inhibited CL-induced cytokine secretion. Medium from CL-treated macrophages induced the release of amylase by acinar cells, and Ddt knockdown medium decreased amylase secretion. The target of Z10 was predicted to be ERK2. Z10 increased the thermostability of ERK1/2 but not ERK1 K72A/ERK2 K52A. The docking poses of ERK1 and ERK2 with Z10 were similar. Z10 inhibited ERK1/2 phosphorylation, and Ddt levels and cytokines were regulated by ERK1/2 during AP. Additionally, Z10 could not further inhibit cytokines under ERK1/2 knockdown with CL. Thus, this study revealed that Z10-mediated ERK1/2 inhibition decreased Ddt expression and secretion by macrophages. Ddt inhibition decreased cytokine release and digestive enzyme secretion.

中文翻译：

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吡唑衍生物 Z10 通过抑制 ERK/Ddt 通路改善急性胰腺炎

急性胰腺炎（AP）可导致死亡；然而，AP 没有具体的治疗方法。很少进行 AP 治疗药物的筛选。在原代胰腺腺泡细胞和腹膜巨噬细胞共培养系统中筛选化合物。化合物在体外和体内使用。预测并验证了复合目标。在 18 种含氮杂环化合物中，Z10 可减少雨蛙素加脂多糖 (CL) 诱导的腺泡消化酶和巨噬细胞细胞因子的分泌。Z10 还被证明可以改善 CL 诱导的或牛磺胆酸钠诱导的小鼠 AP。蛋白质组学分析和酶联免疫吸附测定 (ELISA) 显示，在 CL 处理下，Z10 降低了巨噬细胞内和细胞外环境中的 D-多巴色素互变异构酶 (Ddt) 水平。Z10 还降低了 AP 小鼠中 Ddt 的表达。此外，外源Ddt诱导细胞因子和消化酶的分泌，Z10可以抑制这种分泌。Ddt 敲除抑制 CL 诱导的细胞因子分泌。CL 处理的巨噬细胞的培养基诱导腺泡细胞释放淀粉酶，而 Ddt 敲低培养基则减少淀粉酶的分泌。Z10的靶点预计是ERK2。Z10 增加了 ERK1/2 的热稳定性，但没有增加 ERK1 K72A/ERK2 K52A。ERK1和ERK2与Z10的对接姿势相似。Z10抑制ERK1/2磷酸化，AP期间Ddt水平和细胞因子受ERK1/2调节。此外，在用 CL 敲低 ERK1/2 的情况下，Z10 不能进一步抑制细胞因子。因此，这项研究表明，Z10 介导的 ERK1/2 抑制降低了巨噬细胞的 Ddt 表达和分泌。Ddt 抑制减少了细胞因子的释放和消化酶的分泌。