In the present study, IL-10 was unchanged in the vehicle/APAP group, but increased in the SSd/APAP group

In the present study, IL-10 was unchanged in the vehicle/APAP group, but increased in the SSd/APAP group. countries. Saikosaponin d (SSd, Fig. 1A) is considered one of the major active parts isolated and recognized from this plant [6]. In Sprague-Dawley rats, SSd can decrease transforming growth element 1 in the liver and attenuate the development of hepatic fibrosis and carcinogenesis induced by dimethylnitrosamine [7]. Supplementation with SSd only or in combination with curcumin, significantly reduced carbon tetrachloride (CCl4)-induced swelling and fibrogenesis [8]. In cell tradition models, SSd exhibited potent cytotoprotection and anti-proliferation activity against hepatocellular carcinoma cells [9,10]. However, there have been no studies to evaluate the protecting effect Exo1 of SSd against hepatotoxicity induced by APAP. Open in a separate windows Fig. Exo1 1 Structure of and fragmentation pattern of SSd, and levels of serum SSd in the mice treated with SSd 2mg/kg twice daily for 5 days. A: SSd structure and its proposed fragmentation pattern. B: SSd concentration 1 h after administration monitored on day time 1, 3 and 5. C: Relative abundance of major urinary APAP metabolites involved in APAP-induced liver toxicity. Data were determined by normalizing the solitary ion counts of each metabolite the total ion counts of each urine sample (n=5; **[12]. Safety against CCl4-induced swelling and fibrogenesis by SSd was correlated with down-regulation of the pro-inflammatory cytokines tumor necrosis element- (TNF), IL-1, and IL-6, Exo1 and up-regulation of the anti-inflammatory cytokine IL-10 [8]. Despite the risk of APAP-induced toxicity and the wide software of for liver diseases in clinic, there are no data on the effect of or SSd on APAP-induced hepatotoxicity as well as the underlying mechanism. In this study, APAP was injected to SSd-pretreated C57/B6 mice and changes in liver phenotypes Exo1 and gene expression were examined. 2. Materials and Methods 2.1. Chemicals and reagents Saikosaponin Rabbit Polyclonal to MAEA d (SSd, Fig. 1A), APAP, glutathione (GSH) assay kit, and chlorpropamide were purchased from SigmaCAldrich (Sigma-Aldrich, St. Louis, MO). Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) assay kits were from Catachem (Bridgeport, CT). Antibodies against NFB subunit p65 and signal transducer and activator of transcription 3 (STAT3) and their phosphorylated form, p-p65 and p-STAT3, and GAPDH were purchased from Cell Signaling Technologies (Danvers, MA). HPLC grade solvents such as acetonitrile and formic acid were purchased from Fisher Scientific (Hampton, NH). All the other chemicals were of the highest grade from commercial source. 2.2. Animals and drug administration Male 6- to 7-week-old C57BL6 mice (Jackson Laboratories, Bar Harbor, ME) were maintained in the NCI animal facility under a standard 12 h light/12 h dark cycle with free access to food and water. All procedures were performed in accordance with Institute of Laboratory Animal Resource Guidelines and the animal study protocols approved by the National Cancer Institute Animal Care and Use Committee. Mice were randomly divided into four groups, vehicle/control, SSd/control, vehicle/APAP, and SSd/APAP, and killed 4 h or 24 h after single APAP injection. For APAP injection, a typical single dose of 200 mg/kg/day was used as described elsewhere [3,13,14]. Considering the published pharmacodynamic and pharmacokinetic information of SSd [6,7], 2 mg/kg once daily was used as the dosing regimen. SSd powder was Exo1 dissolved in a saline solution supplemented with 0.1% Tween 20 and was administered by intraperitoneal injection at a dose of 2 mg/kg/day once daily for five days. Saline solution made up of 0.1% Tween 20 without SSd was administered as a vehicle. APAP was dissolved in warm saline solution (20 mg/mL) and was injected intraperitoneally 30 minutes after the last SSd injection. Saline was injected.