Phospholipase D2 targeted by miR-5132-5p alleviates cerulein-induced acute pancreatitis via the Nrf2/NFκB pathway

by Selleckchem | Sep 19 2023

Background: Acute pancreatitis (AP) is an inflammatory process unexpectedly occurring in the pancreas, imposing a substantial burden on healthcare systems. Herein, we aimed to clarify the mechanism of action of phospholipase D2 (PLD2) in cerulein-treated AR42J cells, affording valuable insights into the treatment of AP.

Methods: The levels of PLD2, miR-5132-5p, inflammatory factors (interleukin [IL]-10, IL-6, and tumor necrosis factor-α), caspase-3 activity, and apoptosis-related proteins (Bax and Bcl-2) in cerulein-treated AR42J cells were detected using reverse transcription-quantitative polymerase chain, caspase-3 activity, and Western blot analysis. Protein levels of nuclear Factor erythroid 2-Related Factor 2 (Nrf2) and nuclear factor-k-gene binding (NF-κB) were detected by Western blot analysis. TargetScan predicted upstream microRNAs (miRNAs) of PLD2, and the interaction between miR-5132-5p and PLD2 was verified using a luciferase assay.

Results: In cerulein-treated AR42J cells, PLD2 levels were downregulated, while miR-5132-5p expression was upregulated. Overexpression of PLD2 attenuated the cerulein-mediated facilitatory effect on inflammation and apoptosis in AR42J cells by regulating the Nrf2/NFκB pathway. Luciferase reporter analysis revealed that miR-5132-5p targeted PLD2, and miR-5132-5p negatively regulated PLD2. Upregulation of miR-5132-5p expression exacerbated inflammation and apoptosis and reversed the protective effect of PLD2 overexpression on AP.

Conclusion: PLD2 targeted by miR-5132-5p can attenuate cerulein-induced AP in AR42J cells via the Nrf2/NFκB pathway, providing therapeutic targets for patients with AP.

Comments:

The study you mentioned investigated the role of phospholipase D2 (PLD2) and microRNA-5132-5p (miR-5132-5p) in the development of acute pancreatitis (AP) using cerulein-treated AR42J cells as a model. The researchers aimed to understand the underlying mechanisms and identify potential therapeutic targets for AP.

The methods employed in the study included the measurement of various factors and proteins in cerulein-treated AR42J cells. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR), caspase-3 activity assays, and Western blot analysis were used to detect the levels of PLD2, miR-5132-5p, inflammatory factors (interleukin-10 [IL-10], IL-6, and tumor necrosis factor-α), caspase-3 activity, apoptosis-related proteins (Bax and Bcl-2), nuclear Factor erythroid 2-Related Factor 2 (Nrf2), and nuclear factor-k-gene binding (NF-κB). TargetScan, a bioinformatics tool, was used to predict miRNAs that could potentially regulate PLD2, and the interaction between miR-5132-5p and PLD2 was verified using a luciferase assay.

The results of the study demonstrated that in cerulein-treated AR42J cells, the levels of PLD2 were downregulated, while miR-5132-5p expression was upregulated. Overexpression of PLD2 counteracted the pro-inflammatory and pro-apoptotic effects induced by cerulein in AR42J cells by modulating the Nrf2/NF-κB pathway. Luciferase reporter analysis confirmed that miR-5132-5p directly targeted PLD2 and negatively regulated its expression. Increased expression of miR-5132-5p exacerbated inflammation and apoptosis and reversed the protective effect of PLD2 overexpression on AP.

Based on these findings, the study concludes that targeting PLD2, regulated by miR-5132-5p, may attenuate cerulein-induced AP in AR42J cells by modulating the Nrf2/NF-κB pathway. These findings provide potential therapeutic targets for the treatment of patients with AP. It suggests that strategies aimed at upregulating PLD2 or inhibiting miR-5132-5p expression could be explored as therapeutic interventions for AP.