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MiR-325-3p Alleviates Acute Pancreatitis via Targeting RIPK3

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Abstract

Background and Aims

Acute pancreatitis (AP) is an acute inflammatory disease that can lead to death. Mir-325-3p is strongly and abnormally expressed in many diseases, necessitating exploration of its function and mechanism in AP.

Methods

Blood samples from AP patients and mice were analyzed. The expression levels of miR-325-3p in AP patients and mouse were detected. Whether miR-325-3p targets RIPK3 gene was predicted by TargetScan online database and dual luciferase reporter assay. In vitro experiments verified the effect of miR-325-3p overexpression on caerulein-induced MPC83 pancreatic acinar cancer cell line. In vivo experiments verified the effect of overexpression of miR-325-3p on the disease degree of pancreatic tissues in AP mice.

Results

Analysis of blood samples from AP patients and experiments in mice demonstrated that expression of miR-325-3p was significantly reduced during the process of AP in humans and mice. Predicted using the TargetScan online database and through dual luciferase reporter assay detection, miR-325-3p directly targets the RIPK3 gene. In vitro experiments revealed that overexpression of miR-325-3p reversed caerulein-induced apoptosis and necroptosis in MPC83 pancreatic acinar cancer cell line. We used Z-VAD-FMK to assess necroptosis and demonstrated that miR-325-3p targets necroptosis to reduce cell damage. In subsequent experiments in mice, we verified that overexpression of miR-325-3p reduces inflammation, edema, hemorrhage, and necrosis in acute pancreatitis. Characteristic western blot, immunohistochemistry, and transmission electron microscopy results revealed that overexpression of miR-325-3p reduces the severity of acute pancreatitis by inhibiting pancreatic necroptosis in AP mice.

Conclusions

The current research results indicate that miR-325-3p directly targets RIPK3 and exerts a protective role in mouse AP. Necroptosis is still the primary mechanism of RIPK3 regulation. MiR-325-3p inhibits acute pancreatitis by targeting RIPK3-dependent necroptosis, which may represent a novel treatment method for acute pancreatitis.

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Acknowledgments

We thank all members of the laboratory for useful discussion.

Funding

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Authors and Affiliations

  1. Tianjin Medical University, Tianjin, 300070, China

    Ao Jia, Zhen-Wei Yang, Ji-Yu Shi, Jia-Ming Liu & Yun-Feng Cui

  2. Department of Surgery, Tianjin Nankai Hospital, Nankai Clinical School of Medicine, 122 Sanwei Road, Nankai District, Tianjin, 300110, China

    Kun Zhang & Yun-Feng Cui

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  1. Ao Jia
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  4. Jia-Ming Liu
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Contributions

A.J. and ZW.Y. were involved in the conceptualization; A.J. contributed to the methodology; A.J. and JM.L. contributed to the software; JY.S. and K.Z. were involved in the validation; JM.L. contributed to the investigation; JY.S. contributed to resources; A.J. was involved in the data curation; A.J. was involved in the writing—original draft preparation; ZW.Y. contributed to the writing—review and editing; A.J. was involved in the visualization; YF.C. was involved in the supervision; YF.C. was involved in the project administration; YF.C acquired the funding. All authors have read and agreed to the published version of the manuscript.

Corresponding author

Correspondence to Yun-Feng Cui.

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Conflict of interest

The authors declare no conflict of interest in this work.

Ethical statement

Institutional Review Board Statement: This study was approved by the Ethics Committee of Nankai Hospital and was performed in accordance with the Helsinki Declaration of Principles (ethics review item number: NKYY_YXKT_IRB_2020_059_01). All animal experiments were authorized by the Animal Ethics Committee of Tianjin Nankai Hospital. The ethical review item number is NKYY-DWLL-2020-115, and this study was performed in accordance with the National Laboratory Animal Care and Use Guide.

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Informed consent was obtained from all subjects involved in the study.

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Supplementary file 1 Figure S1: Injection flow chart. (TIF 24598 kb)

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Jia, A., Yang, ZW., Shi, JY. et al. MiR-325-3p Alleviates Acute Pancreatitis via Targeting RIPK3. Dig Dis Sci 67, 4471–4483 (2022). https://doi.org/10.1007/s10620-021-07322-6

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  • DOI: https://doi.org/10.1007/s10620-021-07322-6

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