Knockdown of myeloid differentiation protein-2 reduces acute lung injury following orthotopic autologous liver transplantation in a rat model

Pulm Pharmacol Ther. 2013 Jun;26(3):380-7. doi: 10.1016/j.pupt.2013.02.001. Epub 2013 Feb 19.

Abstract

Background: Acute lung injury (ALI) is a serious complication that commonly occurs during orthotopic liver transplantation (OLT). Toll-like receptor 2/4 (TLR2/4) are the main membrane receptors that respond to inflammatory stimuli and mediate NF-kappa B (NF-κB) signal pathway. We previously showed that TLR2/4 expression on monocytes and serum cytokine levels were increased in patients with ALI induced by OLT. Myeloid differentiation protein-2 (MD-2) expresses the functional domains that combines TLRs and play a key regulatory role in TLRs activation. Therefore, we hypothesized that blocking MD-2 would inhibit the TLR2/4-mediated inflammatory response and lessen ALI induced by liver transplantation.

Method: Thirty-two Sprague Dawley (SD) rats were randomly divided into four groups. One group received a sham operation (Group S), and the other three groups underwent orthotopic autologous liver transplantation (OALT) 48 h after intratracheal administration of saline (Model group; Group M), non-targeting siRNA (negative siRNA control group; Group NC) or siRNA against MD-2 (intervention group; Group RNAi). Lung pathology, lung water content, PaO2, and expression levels of MD-2, TLR2/4, NF-κB, TNF-α, IL-1β and IL-6 were assessed 8 h after OALT.

Results: In Groups M and NC, OALT produced marked lung pathology with decreased PaO2 levels and increased MD-2, TLR2/4 gene and protein expression levels. Furthermore, the nuclear translocation of the NF-κB P65 subunit, was increased, as were lung concentrations of TNF-α, IL-1β and IL-6. The pathology of ALI and the severity of the above biochemical changes induced by OALT were significantly reduced in the group treated with MD-2 siRNA.

Conclusion: MD-2 gene knock-down attenuated the increase in TLR2/4 activation and reduced ALI after OALT.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acute Lung Injury / drug therapy*
  • Acute Lung Injury / physiopathology*
  • Animals
  • Disease Models, Animal
  • Extravascular Lung Water / metabolism
  • Gene Expression
  • Inflammation / metabolism
  • Inflammation / pathology
  • Interleukins / biosynthesis
  • Liver Transplantation / adverse effects*
  • Lung / pathology
  • Lymphocyte Antigen 96 / pharmacology*
  • Male
  • NF-kappa B / biosynthesis
  • RNA, Small Interfering / pharmacology
  • Rats
  • Rats, Sprague-Dawley
  • Toll-Like Receptor 2 / biosynthesis
  • Toll-Like Receptor 4 / biosynthesis
  • Tumor Necrosis Factor-alpha / biosynthesis

Substances

  • Interleukins
  • LY96 protein, human
  • Lymphocyte Antigen 96
  • NF-kappa B
  • RNA, Small Interfering
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4
  • Tumor Necrosis Factor-alpha