MiR-339-3p regulated acute pancreatitis induced by caerulein through targeting TNF receptor-associated factor 3 in AR42J cells.

Qi Wang,Shaofeng Liu,Zhen Han

doi:10.1515/biol-2020-0084

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.

Highlights

Acute pancreatitis (AP) is a sudden pancreatic inflammatory response accompanied by strong pain, mainly caused by necrosis of pancreatic acinar cells
3.2 miR-339-3p expression was inhibited while TNF receptorassociated factor 3 (TRAF3) expression was increased in AR42J cells induced by caerulein
The above data indicated that caerulein induced inflammatory response and apoptosis in AR42J cells suggesting that in vitro model of AP was characterized by a strong inflammatory response and high cell apoptosis

Summary

Introduction

Acute pancreatitis (AP) is a sudden pancreatic inflammatory response accompanied by strong pain, mainly caused by necrosis of pancreatic acinar cells. With the development of gene sequencing and biotechnology, the research on the mechanism of AP has been greatly developed. It provides an important theoretical basis for improving treatment methods and developing molecular therapy. MiRNA-related regulatory networks and mechanisms have been reported to play a crucial role in the mechanisms of diseases [4]. Various studies have shown that miRNA, as an important regulator, is involved in cellular processes in a variety of diseases, including GC, breast cancer, HCC, NCSLC, and pancreatitis [6–9]. The high expression of miR-21 in AP promoted apoptotic activity, and the promotion of miR-22 and miR-135a may induce the apoptosis of pancreatic acinar cells by repressing ErbB3 and Ptk expression [10,11]. Growing evidence determined that miRNAs took part in cell progression

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