Eight- to ten-week-old C57BL/6J male mice (weight: 20–25 g) were purchased from Beijing Spelford Laboratory Animal Technology Co., Ltd. (China). A cecal ligation perforation operation was used to prepare an animal model of sepsis. C57BL/6J mice were randomly divided into five groups: (I) sham-operated group (sham group, n=8); (II) cecum ligation perforation group (CLP group, n=8); (III) CLP + liprestatin-1 (Abcam, Cambridge, UK) group (CLP + Lip-1 group, n=8); (IV) CLP + recombinant human Klotho protein (Abcam) group (CLP + Klotho group, n=8); and (V) CLP + Klotho + Nrf2-IN-1 (GlpBio, Montclair, CA, USA) group (n=8).
Mice were injected with Lip-1 (10 mg/kg) and Klotho protein (10 mg/kg) intraperitoneally 1 h before the construction of the CLP model; Nrf2-IN-1 (10 mg/kg) was injected intraperitoneally 12 h before the construction of the CLP model. Mice were anesthetized with isoflurane (induction concentration 3.5%, maintenance concentration 3%). Experiments were performed under a project license (No. 2022006) granted by Animal Experiment Center of the Fourth Hospital of Hebei Medical University, in compliance with the National Institutes of Health (USA) Guidelines for the care and use of animals (NIH Publication No. 85-23, revised 1985:http://grants1.nih.gov/grants/olaw/references/phspol.htm ). A protocol was prepared before the study without registration.
Mice were injected with Lip-1 (10 mg/kg) and Klotho protein (10 mg/kg) intraperitoneally 1 h before the construction of the CLP model; Nrf2-IN-1 (10 mg/kg) was injected intraperitoneally 12 h before the construction of the CLP model. Mice were anesthetized with isoflurane (induction concentration 3.5%, maintenance concentration 3%). Experiments were performed under a project license (No. 2022006) granted by Animal Experiment Center of the Fourth Hospital of Hebei Medical University, in compliance with the National Institutes of Health (USA) Guidelines for the care and use of animals (NIH Publication No. 85-23, revised 1985: