Department of Medicine, Division of Gastroenterology
Animals; Carrier Proteins; Cytokines; Inflammasomes; Lipopolysaccharides; Liver; Mice; Mice, Inbred C57BL
Amino Acids, Peptides, and Proteins | Animal Experimentation and Research | Biological Factors | Carbohydrates | Digestive System | Enzymes and Coenzymes | Gastroenterology | Investigative Techniques | Lipids | Macromolecular Substances
AIM: To examine the activation of the Nalp3 inflammasome and its downstream targets following lipopolysaccharide (LPS)-induced stimulation in the liver.
METHODS: Six-to-eight-week-old C57BL/6 chow fed mice were injected intraperitoneally with 0.5 mug/g bodyweight LPS and sacrificed 2, 4, 6, 18 or 24 h later. LPS-induced liver damage was confirmed by a biochemical assay to detect alanine aminotransferase (ALT) levels. To determine if LPS stimulation in the liver led to activation of the inflammasome, real-time quantitative polymerase chain reaction was used to evaluate the mRNA expression of components of the Nalp3 inflammasome. Enzyme-linked immunosorbent assays were used to determine the protein expression levels of several downstream targets of the Nalp3 inflammasome, including caspase-1 and two cytokine targets of caspase-1, interleukin (IL)-1beta and IL-18.
RESULTS: We found that LPS injection resulted in liver damage as indicated by elevated ALT levels. This was associated with a significant increase in both mRNA and protein levels of the proinflammatory cytokine tumor necrosis factor (TNF)-alpha in the liver, as well as increased levels of TNFs in serum. We showed that LPS stimulation led to upregulation of mRNA levels in the liver for all the receptor components of the inflammasome, including Nalp3, Nalp1, pannexin-1 and the adaptor molecule apoptosis-associated speck-like, caspase recruitment domain-domain containing protein. We also found increased levels of mRNA and protein for caspase-1, a downstream target of the inflammasome. In addition, LPS challenge led to increased levels of both mRNA and protein in the liver for two cytokine targets of caspase-1, IL-1beta and IL-18. Interestingly, substantial baseline expression of pre-IL-1beta and pre-IL-18 was found in the liver. Inflammasome and caspase-1 activation was indicated by the significant increase in the active forms of IL-1beta and IL-18 after LPS stimulation.
CONCLUSION: Our results show that the Nalp3 inflammasome is upregulated and activated in the liver in response to LPS stimulation.
Endotoxin, Nod-like receptor, Interleukin- 1β, Interleukin-18, Caspase-1
DOI of Published Version
World J Gastroenterol. 2011 Nov 21;17(43):4772-8. Link to article on publisher's site
World journal of gastroenterology : WJG
Ganz, Michal; Csak, Timea; Nath, Bharath D.; and Szabo, Gyongyi, "Lipopolysaccharide induces and activates the Nalp3 inflammasome in the liver" (2011). University of Massachusetts Medical School. Senior Scholars Program. Paper 135.
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