Kallistatin treatment attenuates lethality and organ injury in mouse models of established sepsis.
BACKGROUND
Kallistatin levels in the circulation are reduced in patients with sepsis and liver disease. Transgenic mice expressing kallistatin are resistant to lipopolysaccharide (LPS)-induced mortality. Here, we investigated the effect of kallistatin on survival and organ damage in mouse models of established sepsis.


METHODS
Mice were rendered septic by cecal ligation and puncture (CLP), or endotoxemic by LPS injection. Recombinant human kallistatin was administered intravenously six hours after CLP, or intraperitoneally four hours after LPS challenge. The effect of kallistatin treatment on organ damage was examined one day after sepsis initiation, and mouse survival was monitored for four to six days.


RESULTS
Human kallistatin was detected in mouse serum of kallistatin-treated mice. Kallistatin significantly reduced CLP-induced renal injury as well as blood urea nitrogen, serum creatinine, interleukin-6 (IL-6), and high mobility group box-1 (HMGB1) levels. In the lung, kallistatin decreased malondialdehyde levels and HMGB1 and toll-like receptor-4 (TLR4) synthesis, but increased suppressor of cytokine signaling-3 (SOCS3) expression. Moreover, kallistatin attenuated liver injury, serum alanine transaminase (ALT) levels and hepatic tumor necrosis factor-α (TNF-α) synthesis. Furthermore, delayed kallistatin administration improved survival in CLP mice by 38%, and LPS-treated mice by 42%. In LPS-induced endotoxemic mice, kallistatin attenuated kidney damage in association with reduced serum creatinine, IL-6 and HMGB1 levels, and increased renal SOCS3 expression. Kallistatin also decreased liver injury in conjunction with diminished serum ALT levels and hepatic TNF-α and TLR4 expression. In cultured macrophages, kallistatin through its active site increased SOCS3 expression, but this effect was blocked by inhibitors of tyrosine kinase, protein kinase C and extracellular signal-regulated kinase (ERK), indicating that kallistatin stimulates a tyrosine-kinase-protein kinase C-ERK signaling pathway.


CONCLUSIONS
This is the first study to demonstrate that delayed human kallistatin administration is effective in attenuating multi-organ injury, inflammation and mortality in mouse models of polymicrobial infection and endotoxemia. Thus, kallistatin therapy may provide a promising approach for the treatment of sepsis in humans.
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, 173 Ashley Ave, Charleston, SC, 29425-2211, USA. lippe@musc.edu.
Pengfei
P
Li
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, 173 Ashley Ave, Charleston, SC, 29425-2211, USA. guoy@musc.edu.
Youming
Y
Guo
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, 173 Ashley Ave, Charleston, SC, 29425-2211, USA. jandgbledsoe@bellsouth.net.
Grant
G
Bledsoe
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, 173 Ashley Ave, Charleston, SC, 29425-2211, USA. yangz@musc.edu.
Zhi-Rong
ZR
Yang
Department of Neurosciences, Medical University of South Carolina, 173 Ashley Ave, Charleston, SC, 29425-2211, USA. fanhong@musc.edu.
Hongkuan
H
Fan
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, 173 Ashley Ave, Charleston, SC, 29425-2211, USA. chaol@musc.edu.
Lee
L
Chao
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, 173 Ashley Ave, Charleston, SC, 29425-2211, USA. chaoj@musc.edu.
Julie
J
Chao
D058186:Acute Kidney Injury / Q000517:prevention & control*; D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D001806:Blood Urea Nitrogen; D002478:Cells, Cultured; D003404:Creatinine / Q000097:blood; D004195:Disease Models, Animal; D019446:Endotoxemia / Q000378:metabolism; D024243:HMGB1 Protein / Q000378:metabolism; D006801:Humans; D015850:Interleukin-6 / Q000097:blood; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D008315:Malondialdehyde / Q000378:metabolism; D051379:Mice; D011994:Recombinant Proteins / Q000494:pharmacology*; D018805:Sepsis / Q000188:drug therapy* / Q000378:metabolism; D015843:Serpins / Q000097:blood / Q000494:pharmacology*; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D024243:HMGB1 Protein; C582808:HMGB1 protein, mouse; D015850:Interleukin-6; D011994:Recombinant Proteins; D015843:Serpins; C491411:Socs3 protein, mouse; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; C051324:kallistatin; D008315:Malondialdehyde; D003404:Creatinine; D000410:Alanine Transaminase