Lipopolysaccharide lps escherichia coli 055 b5

Phosphate-buffered saline (PBS) and Dulbecco’s Modified Eagle’s Medium (DMEM) was obtained from Corning, Inc. (New York, NY, USA)., fetal bovine serum (FBS) were obtained from Gibco (Grand Island, CA, USA). Penicillin-Streptomycin was Thermo fisher (Waltham, MA, USA). Lipopolysaccharide (LPS, Escherichia coli 055:B5) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies against rabbit-anti COX-2, NF-κB p65, ERK, phospho-extracellular signal-regulated kinase (p-ERK), p38, p-p38, JNK, phospho-Jun N-terminal (p-JNK), phosphoinositide 3-kinase (PI3K), p-AKT, AKT, and Lamin B1 were purchased from Cell Signaling Technology (Beverly, MA, USA). Mouse anti-iNOS was purchased from from R&D Systems Inc.(Minneapolis, MN, USA). IL-1β and TNF-α were purchased from Cruise Biotechnology (Santa Cruz, CA, USA). Mouse anti-β-actin and goat anti-rabbit IgG HRP antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

C57BL/6 mice (male, 18–23 g; 6 weeks old) were obtained from OrientBio Co., Ltd. and housed in a temperature‐controlled room at 21 ± 2°C and 50 ± 20% humidity with a 12‐hr light/dark cycle. Mice had free access to standard laboratory food and water ad libitum. Before the experiments, they were acclimatized for 1 week. All animal experiments were approved by the Animal Research Ethics Committee of KPC (Approval Number: P173010; Gwangju, Gyeonggi‐do, Korea). Lipopolysaccharide (LPS; Escherichia coli, 055: B5) and D‐galactosamine (D‐galN) were purchased from Sigma‐Aldrich.
Fifty mice were divided into five groups: (a) untreated naïve control group, (b) LPS/D‐galN‐treated liver failure group, (c) LPS/D‐galN + SME: Silybum marianum extract (100 mg/ml b.w.t) group, (d) LPS/D‐galN + AAWE (100 mg/ml b.w.t) group, and (e) LPS/D‐galN + AAEE (100 mg/ml b.w.t) group. Test materials were administrated orally once a day for 14 days. After 1 hr of the last oral administration, LPS (4 μg/kg) and D‐galN (400 mg/kg) were administrated by intraperitoneal injection. Blood, liver tissues, and spleen tissues were collected for biochemical and histological analyses after LPS/D‐galN treatment for 4 hr.