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Regulation of Postoperative Ileus by Lentivirus-Mediated HuR RNA Interference via the p38/MK2 Signaling Pathway

  • Original Article
  • Published:
Journal of Gastrointestinal Surgery

Abstract

Objectives

The objective of the present study is to explore the effect of lentivirus-mediated HuR interference on the development and progression of postoperative ileus and the role of HuR in the regulation of the p38/MAPK-activated protein kinase-2 (MK2) signaling pathway during postoperative ileus.

Methods

To establish a mouse model of lentiviral transduction, we first determined the optimum effective titer of lentiviral vectors for transduction of the murine small intestine via the abdominal cavity by using hematoxylin and eosin (HE) staining, immunohistochemistry, detection of GFP messenger RNA (mRNA) and protein, and Western blotting. To investigate the effect of HuR interference on gene expression during postoperative ileus, we established a mouse model of postoperative ileus and used RT-PCR to measure the expression of proinflammatory genes, ELISA to measure the expression of serum inflammatory cytokines, immunohistochemistry to evaluate inflammatory cell infiltration in the small intestine, HE staining of paraffin sections to examine the pathology of the small intestine, and Western blotting to measure HuR expression and identify its role in the regulation of the p38/MK2 inflammatory pathway.

Results

We successfully designed a mouse model of intraperitoneal transduction of HuR-RNAi lentivirus. When HuR gene expression was suppressed in a mouse model of postoperative ileus, the infiltration of inflammatory cells, the expression of proinflammatory genes, and the levels of serum inflammatory cytokines were significantly reduced. This reduction in inflammation correlated with reduced cytoplasmic localization of HuR and reduced activation of MK2.

Conclusions

Within the p38/MK2 signal transduction pathway, HuR may increase the mRNA stability of various inflammatory cytokines, thereby promoting inflammation that causes postoperative ileus. Suppressing the expression of HuR in a postoperative ileus model can effectively suppress the postoperative ileus inflammatory reaction. HuR might serve as a candidate drug target for the prevention and mitigation of postoperative ileus.

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Guarantor of the Article

Chi Pan accepts full responsibility for the conduct of the study, has had access to the data, and has control of the decision to publish.

Specific Author Contributions

Ye Dao Xiong and Chi Pan conceived the study, collected and analyzed the data, and drafted the manuscript. Lu Xing Rong critically reviewed and revised the manuscript. All authors read and approved the final manuscript.

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Authors and Affiliations

  1. Fujian Medical University Union Hospital, Fuzhou, China

    Ye Dao Xiong, Lu Xing Rong & Chi Pan

Authors
  1. Ye Dao Xiong
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  2. Lu Xing Rong
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  3. Chi Pan
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Corresponding author

Correspondence to Ye Dao Xiong.

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Financial Support

This study was supported by the National Key Clinical Specialty Discipline Construction Program of China (No. 2015-8147277) and the Key Project of Science and Technology Plan of Fujian Province, China (No. 2011J01172 and 2012J01349).

Conflict of Interest

The authors declare that they have no conflicts of interest.

Additional information

Study Highlights

1. What is the current knowledge?

• Currently, there is no effective method to prevent postoperative ileus.

• Inflammatory response arising 3–4 h postoperation is the leading cause of postoperative ileus.

• There is no targeted therapy that prevents postoperative inflammatory responses.

2. What is new here?

• HuR promotes postoperative inflammation by stabilizing mRNAs of inflammatory cytokines.

• Blocking HuR expression ameliorates inflammation in a mouse model of postoperative ileus.

• HuR may provide a potential therapeutic target to prevent and reduce postoperative ileus.

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Xiong, Y.D., Rong, L.X. & Pan, C. Regulation of Postoperative Ileus by Lentivirus-Mediated HuR RNA Interference via the p38/MK2 Signaling Pathway. J Gastrointest Surg 21, 389–397 (2017). https://doi.org/10.1007/s11605-016-3303-z

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  • DOI: https://doi.org/10.1007/s11605-016-3303-z

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