Abstract Acute pancreatitis (AP) is an inflammatory disease with high morbidity and mortality. The regulation mechanism of miRNA is involved in the production and development of various diseases, but the regulation mechanism of miRNA in AP is still not fully elucidated. The expression of miR-339-3p was detected using quantitative real-time PCR. The levels of TNF-α, IL-1β, and IL-6 were detected using enzyme-linked immunosorbent assay. Cell apoptosis was measured using flow cytometry. The protein expressions of TNF receptor-associated factor 3 (TRAF3), Bcl-2, C-caspase 3, Bax, p-p38, and p38 were measured using western blot. Luciferase reporter assay and RNA immunoprecipitation assay were applied to ensure that miR-399-3p targeted TRAF3. Caerulein promoted the expression of TNF-α, IL-1β, and IL-6, enhanced the expression of C-caspase 3 and Bax while inhibited Bcl-2 protein expression. Meanwhile, caerulein also reduced the expression of miR-339-3p and induced the expression of TRAF3 in rat pancreatic acinar cells. miR-399-3p transfection inhibited the levels of TNF-α, IL-1β, and IL-6 and C-caspase 3 and Bax protein expression as well as suppressed cell apoptosis, while increased Bcl-2 protein expression in caerulein-induced AP. TRAF3 has been verified as a target of miR-339-3p. Interestingly, the reduction of miR-399-3p inhibited the p38 pathway, which was impaired by the upregulation of TRAF3. In addition, the suppression effects of miR-339-3p on cell inflammation and apoptosis in caerulein-induced AP were reversed by enhancing TRAF3 expression. In this study, in vitro model of AP was characterized by strong inflammation and cell apoptosis. We have first demonstrated the regulatory network of miR-339-3p and TRAF3. Overexpression of miR-339-3p inhibited cell inflammation and cell apoptosis in caerulein-induced AP through modulating TRAF3 expression via the p38 pathway, providing a new therapeutic target in the treatment of AP.

中文翻译：

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miR-339-3p 通过靶向 AR42J 细胞中的 TNF 受体相关因子 3 调控雨蛙素诱导的急性胰腺炎

摘要 急性胰腺炎（AP）是一种发病率和死亡率都很高的炎症性疾病。miRNA的调控机制参与了多种疾病的产生和发展，但miRNA在AP中的调控机制尚未完全阐明。使用定量实时PCR检测miR-339-3p的表达。使用酶联免疫吸附法检测TNF-α、IL-1β和IL-6的水平。使用流式细胞术测量细胞凋亡。使用蛋白质印迹法测量 TNF 受体相关因子 3 (TRAF3)、Bcl-2、C-半胱天冬酶 3、Bax、p-p38 和 p38 的蛋​​白质表达。应用荧光素酶报告基因测定和 RNA 免疫沉淀测定以确保 miR-399-3p 靶向 TRAF3。Caerulein 促进 TNF-α、IL-1β 和 IL-6 的表达，增强 C-caspase 3 和 Bax 的表达，同时抑制 Bcl-2 蛋白的表达。同时，雨蛙素还降低了miR-339-3p的表达，并诱导了大鼠胰腺腺泡细胞中TRAF3的表达。miR-399-3p 转染抑制 TNF-α、IL-1β、IL-6 和 C-caspase 3 和 Bax 蛋白的表达水平并抑制细胞凋亡，同时增加了雨蛙素诱导的 AP 中 Bcl-2 蛋白的表达. TRAF3 已被证实为 miR-339-3p 的靶点。有趣的是，miR-399-3p 的减少抑制了 p38 通路，该通路因 TRAF3 的上调而受损。此外，通过增强 TRAF3 表达，miR-339-3p 对金盏花素诱导的 AP 中细胞炎症和细胞凋亡的抑制作用被逆转。在本研究中，AP体外模型的特征是强烈的炎症和细胞凋亡。我们首先展示了 miR-339-3p 和 TRAF3 的调控网络。miR-339-3p的过表达通过p38通路调节TRAF3的表达，抑制了雨蛙素诱导的AP中的细胞炎症和细胞凋亡，为AP的治疗提供了新的治疗靶点。