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Molecular Medicine

, Volume 18, Issue 1, pp 29–37 | Cite as

Serum from Patients Undergoing Remote Ischemic Preconditioning Protects Cultured Human Intestinal Cells from Hypoxia-Induced Damage: Involvement of Matrixmetalloproteinase-2 and -9

  • Karina Zitta
  • Patrick Meybohm
  • Berthold Bein
  • Christin Heinrich
  • Jochen Renner
  • Jochen Cremer
  • Markus Steinfath
  • Jens Scholz
  • Martin Albrecht
Research Article

Abstract

Remote ischemic preconditioning (RIPC) can be induced by transient occlusion of blood flow to a limb with a blood pressure cuff and exerts multiorgan protection from ischemia/reperfusion injury. Ischemia/reperfusion injury in the intestinal tract leads to intestinal barrier dysfunction and can result in multiple organ failure. Here we used an intestinal cell line (CaCo-2) to evaluate the effects of RIPC-conditioned patient sera on hypoxia-induced cell damage in vitro and to identify serum factors that mediate RIPC effects. Patient sera (n = 10) derived before RIPC (T0), directly after RIPC (T1) and 1 h after RIPC (T2) were added to the culture medium at the onset of hypoxia until 48 h after hypoxia. Reverse transcription-polymerase chain reaction, lactate dehydrogenase (LDH) assays, caspase-3/7 assays, silver staining, gelatin zymography and Western blotting were performed. Hypoxia led to morphological signs of cell damage and increased the release of LDH in cultures containing sera T0 (P < 0.01) and T1 (P < 0.05), but not sera T2, which reduced the hypoxia-mediated LDH release compared with sera T0 (P < 0.05). Gelatin zymography revealed a significant reduction of activities of the matrixmetalloproteinase (MMP)-2 and MMP-9 in the protective sera T2 compared with the nonprotective sera T0 (MMP-2: P < 0.01; MMP-9: P < 0.05). Addition of human recombinant MMP-2 and MMP-9 to MMP-deficient culture media increased the sensitivity of CaCo-2 cells to hypoxia-induced cell damage (P < 0.05), but did not result in a reduced phosphorylation of prosurvival kinases p42/44 and protein kinase B (Akt) or increased activity of caspase-3/7. Our results suggest MMP-2 and MMP-9 as currently unknown humoral factors that may be involved in RIPC-mediated cytoprotection in the intestine.

Notes

Acknowledgments

We thank O Broch, H Franksen, A Carstens, M Betz, C Rodde, M Jonigkeit, I Möller, D Maahs, T Schuett, F Lauer and S Schroeder for technical assistance.

Supplementary material

10020_2012_1801029_MOESM1_ESM.pdf (496 kb)
Serum from Patients Undergoing Remote Ischemic Preconditioning Protects Cultured Human Intestinal Cells from Hypoxia-Induced Damage: Involvement of Matrixmetalloproteinase-2 and -9

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Authors and Affiliations

  • Karina Zitta
    • 1
  • Patrick Meybohm
    • 1
  • Berthold Bein
    • 1
  • Christin Heinrich
    • 1
  • Jochen Renner
    • 1
  • Jochen Cremer
    • 2
  • Markus Steinfath
    • 1
  • Jens Scholz
    • 1
  • Martin Albrecht
    • 1
  1. 1.Department of Anesthesiology and Intensive Care MedicineUniversity Hospital Schleswig-HolsteinKielGermany
  2. 2.Department of Heart and Vascular SurgeryUniversity Hospital Schleswig-HolsteinKielGermany

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