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Cystathionine-gamma-lyase gene silencing with siRNA in monocytes/macrophages protects mice against acute pancreatitis

  • Applied microbial and cell physiology
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Abstract

Hydrogen sulphide (H2S) is an endogenous inflammatory mediator produced by cystathionine-γ-lyase (CSE) in monocytes/macrophages. To determine the role of H2S and macrophages in inflammation, we used small interference RNA (siRNA) to target the CSE gene and investigated its effect in a mouse model of acute pancreatitis. Acute pancreatitis is characterised by increased levels of plasma amylase, myeloperoxidase (MPO) activity and pro-inflammatory cytokines and chemokines in the pancreas and lung. SiRNA treatment attenuated inflammation in the pancreas and lungs of mice following caerulein-induced acute pancreatitis. MPO activity increased in caerulein-induced acute pancreatitis (16.21 ± 3.571 SD fold increase over control) and treatment with siRNA significantly reduced this (mean 3.555 ± 2.522 SD fold increase over control) (p < 0.0001). Similarly, lung MPO activity increased following treatment with caerulein (3.56 ± 0.941 SD fold increase over control) while siRNA treatment significantly reduced MPO activity (0.8243 ± 0.4353 SD fold increase over control) (p < 0.0001). Caerulein treatment increased plasma amylase activity (7094 ± 207 U/l) and this significantly decreased following siRNA administration (5895 ± 115 U/l) (p < 0.0001). Cytokine and chemokine levels in caerulein-induced acute pancreatitis reduced following treatment with siRNA. For example, siRNA treatment significantly decreased pancreatic and lung monocyte chemoattractant protein (MCP)-1 (169.8 ± 59.75 SD; 90.01 ± 46.97 SD pg/ml, respectively) compared to caerulein-treated mice (324.7 ± 103.9 SD; 222.8 ± 85.37 SD pg/ml, pancreas and lun,g respectively) (p < 0.0001). These findings show a crucial pro-inflammatory role for H2S synthesised by CSE in macrophages in acute pancreatitis and suggest CSE gene silencing with siRNA as a potential therapeutic approach for this condition.

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Acknowledgments

This work was supported by the Maurice and Phyllis Paykel Trust grant number 110443.01.Q.PZ and Infection Research Group (Department of Pathology, University of Otago, Christchurch) (Professor Stephen Chambers).

Conflict of interest

The authors declare that they have no competing interests.

Compliance with ethical standards

All experimental procedures were approved by the animal ethics committee of the University of Otago, Christchurch, New Zealand.

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Authors and Affiliations

  1. Inflammation Research Group, Department of Pathology, University of Otago, 2 Riccarton Avenue, P.O. Box 4345, Christchurch, 8140, New Zealand

    A. Badiei, R. R. Gaddam, R. Fraser & M. Bhatia

  2. Infection Research Group, Department of Pathology, University of Otago, Christchurch, New Zealand

    S. T. Chambers

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  1. A. Badiei
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  2. S. T. Chambers
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  3. R. R. Gaddam
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  4. R. Fraser
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  5. M. Bhatia
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Correspondence to M. Bhatia.

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Badiei, A., Chambers, S.T., Gaddam, R.R. et al. Cystathionine-gamma-lyase gene silencing with siRNA in monocytes/macrophages protects mice against acute pancreatitis. Appl Microbiol Biotechnol 100, 337–346 (2016). https://doi.org/10.1007/s00253-015-6989-z

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  • DOI: https://doi.org/10.1007/s00253-015-6989-z

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