CLP-Induced Sepsis Model in Mice

Sepsis was induced by CLP. Briefly, the peritoneal cavity was opened under inhaled isoflurane anesthesia. The cecum was eviscerated, ligated below the ileocecal valve using a 5-0 silk suture at 75% from the tip, and punctured through with a 21-gauge needle. Sham-operated animals were handled in the similar manner, except the cecum was not ligated and punctured. For the survival study, mice were monitored for 7 days. For mechanistic studies, mice were sacrificed at their respective time point. For inhibitor analysis, PAD4 inhibitor GSK484 (40 mg/kg) (Cayman Chemical, 17488), PAD2 inhibitor AFM32a (20 mg/kg), which was synthesized as previously described (65 (link)), or DMSO alone was i.p. injected 1 hour after CLP.

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Tian Y., Qu S., Alam H.B., Williams A.M., Wu Z., Deng Q., Pan B., Zhou J., Liu B., Duan X., Ma J., Mondal S., Thompson P.R., Stringer K.A., Standiford T.J, & Li Y. (2020). Peptidylarginine deiminase 2 has potential as both a biomarker and therapeutic target of sepsis. JCI Insight, 5(20), e138873.

Publication 2020

GSK484

Anesthesia Animals Cayman Cecum Dmso Ileocecal valve Isoflurane Mice Needle Peritoneal cavity Sepsis Silk Suture

Corresponding Organization : Michigan Medicine

Other organizations : Xiangya Hospital Central South University, Central South University, Xijing Hospital, Second Xiangya Hospital of Central South University, Peking University People's Hospital, Peking University, Loyola University Medical Center, The Ohio State University, University of Massachusetts Chan Medical School

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Variable analysis

independent variables

Induction of sepsis by cecal ligation and puncture (CLP)
Administration of PAD4 inhibitor GSK484 (40 mg/kg)
Administration of PAD2 inhibitor AFM32a (20 mg/kg)
Administration of DMSO

dependent variables

Survival of mice over 7 days
Outcomes measured at respective time points for mechanistic studies

control variables

Sham-operated animals (cecum not ligated and punctured)
Inhaled isoflurane anesthesia
Handling and procedures performed in a similar manner for sham-operated animals

controls

Positive control: Sham-operated animals
Negative control: DMSO administration

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