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Molecular Medicine

, Volume 23, Issue 1, pp 155–165 | Cite as

Myosin Light Chain Kinase Knockout Improves Gut Barrier Function and Confers a Survival Advantage in Polymicrobial Sepsis

  • C. Adam Lorentz
  • Zhe Liang
  • Mei Meng
  • Ching-Wen Chen
  • Benyam P. Yoseph
  • Elise R. Breed
  • Rohit Mittal
  • Nathan J. Klingensmith
  • Alton B. Farris
  • Eileen M. Burd
  • Michael Koval
  • Mandy L. Ford
  • Craig M. Coopersmith
Research Article

Abstract

Sepsis-induced intestinal hyperpermeability is mediated by disruption of the epithelial tight junction, which is closely associated with the perijunctional actin-myosin ring. Myosin light chain kinase (MLCK) phosphorylates the myosin regulatory light chain, resulting in increased permeability. The purpose of this study was to determine whether genetic deletion of MLCK would alter gut barrier function and survival from sepsis. MLCK−/− and wild-type (WT) mice were subjected to cecal ligation and puncture and assayed for both survival and mechanistic studies. Survival was significantly increased in MLCK−/− mice (95% versus 24%, p < 0.0001). Intestinal permeability increased in septic WT mice compared with unmanipulated mice. In contrast, permeability in septic MLCK−/− mice was similar to that seen in unmanipulated animals. Improved gut barrier function in MLCK−/− mice was associated with increases in the tight junction mediators ZO-1 and claudin 15 without alterations in claudin 1, 2, 3, 4, 5, 7, 8 and 13, occludin or JAM-A. Other components of intestinal integrity (apoptosis, proliferation and villus length) were unaffected by MLCK deletion, as were local peritoneal inflammation and distant lung injury. Systemic IL-10 was decreased greater than 10-fold in MLCK−/− mice; however, survival was similar between septic MLCK−/− mice given exogenous IL-10 or vehicle. These data demonstrate that deletion of MLCK improves survival following sepsis, associated with normalization of intestinal permeability and selected tight junction proteins.

Notes

Acknowledgments

This work was supported by funding from the National Institutes of Health (GM072808, GM095442, GM104323, GM109779, GM113228, HL116958).

Supplementary material

10020_2017_2301155_MOESM1_ESM.pdf (160 kb)
Supplementary material, approximately 160 KB.

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© The Author(s) 2017

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Authors and Affiliations

  • C. Adam Lorentz
    • 1
  • Zhe Liang
    • 2
  • Mei Meng
    • 3
  • Ching-Wen Chen
    • 2
  • Benyam P. Yoseph
    • 2
  • Elise R. Breed
    • 2
  • Rohit Mittal
    • 2
  • Nathan J. Klingensmith
    • 2
  • Alton B. Farris
    • 4
  • Eileen M. Burd
    • 4
  • Michael Koval
    • 5
  • Mandy L. Ford
    • 6
  • Craig M. Coopersmith
    • 2
    • 7
  1. 1.Department of UrologyEmory University School of MedicineAtlantaUSA
  2. 2.Department of Surgery and Emory Critical Care CenterEmory University School of MedicineAtlantaUSA
  3. 3.Department of Critical Care MedicineShandong Provincial Hospital Affiliated Shandong UniversityJinanChina
  4. 4.Department of Pathology and Laboratory MedicineEmory University School of MedicineAtlantaUSA
  5. 5.Department of Internal Medicine and Emory Alcohol and Lung Biology CenterEmory University School of MedicineAtlantaUSA
  6. 6.Department of Surgery and Emory Transplant CenterEmory University School of MedicineAtlantaUSA
  7. 7.AtlantaUSA

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