Data are presented as the means SD for 6 animals in each group. of liver damage induced by AA within a short-term period. The subcutaneous administration of AA induced severe hepatic injury, and combined treatment with AA and MH resulted in a significant improvement in all evaluated parameters. This recovery was most obvious in the group receiving AA and 15 mg/kg body weight dose of MH. The findings of our study exhibited that MH guarded mice from severe hepatic injury induced by AA. Moreover, MH is a natural polyphenolic compound, and thus it has potential for use in the treatment of severe liver diseases, in place of many synthetic drugs. was shown to protect the livers of mice against chemically and immunologically-induced injury (3). Thus, it is important to discover methods with which to ameliorate the pathological symptoms of liver disease in order to prevent and treat liver injury. Natural polyphenolic compounds extracted from traditional plants are considered safe and effective alternative treatments to alleviate the symptoms of hepatotoxicity; for example, hydroethanolic extracts have been shown to effectively alleviate acetaminophen-induced hepatotoxicity in rats (4). Current research into hepatoprotective natural compounds is a major indication for the screening systems that can trigger Fomepizole a security evaluation in the early phase of drug discovery as the majority of toxic compounds are metabolized in the liver. Acrylamide (AA) is usually a chemical compound that is widely used in the synthesis of polyacrylamides and other industrial chemicals. It has been demonstrated to induce oxidative stress, genotoxicity, neurotoxicity, carcinogenicity, developmental disorders and reproductive toxicity in rodents and other animals (5,6). These effects are mediated through two major metabolic pathways: in the first pathway, the conjugation of AA with glutathione forms N-acetyl-S-(3-amino-3-oxopropyl) cysteine and N-acetyl-S-(2-carbamoylethyl) cysteine, and in the second pathway, the epoxidation of AA into glycidamide occurs via cytochrome (Cyt)P450 (7), and the biotransformation of AA to its epoxide derivative prospects to the formation of glycidamide-DNA adducts, which leads to DNA damage (8,9). Studies have investigated the use of real natural compounds to control AA-induced toxicity, by regulating the levels of CytP450 and poly(ADP-ribose) polymerase (PARP)-1 (10,11). Recently, there has been a growing desire for exploiting natural real compounds due to their natural origin, affordable cost and minimal side-effects. Of these, morin hydrate (MH; 2,3,4,5,7-pentahydroxyflavone), a yellowish pigment and potent flavonoid abundantly present in the plants of the Moraceae family (Fig. 1), is usually thought to be a major bioactive compound that may be used to prevent hepatotoxicity (12,13). The protective Fomepizole effects of MH against oxidative stress and inflammation, which are the result of scavenging oxygen free radicals and hydroxyl radicals, have been investigated in previous studies (14C16). In addition, MH exerts numerous beneficial pharmacological effects, including the inhibition of xanthine oxidase (17), the prevention of low-density lipoprotein oxidation (18), immunomodulation (19) and anticancer activity (20). Open in a separate window Physique 1 Chemical structure of morin hydrate. In light of the aforementioned medical properties of MH, in the present study, we thus aimed to investigate the hypothesis that MH may protect the liver against injury due to AA-induced toxicity by modulating the expression of inducible nitric oxide synthase (iNOS), endothelial nitric oxide Rabbit Polyclonal to ENTPD1 synthase (eNOS), CytP450, PARP-1, antioxidant enzymes and other pathological parameters related to hepatotoxicity in a male murine model. Materials and methods Reagents MH and AA were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-rabbit PARP-1 (sc7150; Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), anti-CytP450 (3084R100; BioVision, Inc., Milpitas, CA, USA), anti-eNOS (ab66127; Abcam, Cambridge, MA, USA), anti-iNOS (ab3523; Abcam), anti-catalase (sc50508; Santa Cruz Biotechnology Inc.), anti-glutathione reductase (GR; sc32886; Santa Cruz Biotechnology Inc.), anti-superoxide dismutase (SOD)1 (sc-11407; Santa Cruz Biotechnology Inc.), anti-SOD2 (sc-30080; Santa Cruz Biotechnology Inc.) and anti–actin (sc-1616; Santa Cruz Biotechnology Inc.) antibodies, and horseradish peroxidase (HRP)-conjugated anti-rabbit immunoglobulins were purchased from Santa Cruz Biotechnology Inc. All chemicals, Fomepizole unless otherwise stated, were of the highest quality and were purchased from Sigma-Aldrich. Animals and experimental design Four-week-old male ICR mice, each weighing 205 g, were provided by Orient Bio Inc. (Seongnam, Korea). All research procedures were carried out in accordance with the Guidelines for Animal Experimentation of Daegu University or college (Daegu, Korea). The animals were housed in an air-conditioned room at 222C and 3010% relative humidity. The animals were observed in terms of their general condition during a 7-day quarantine and acclimation period to confirm there were no abnormalities. After the quarantine period, the mice were randomly divided into 4 groups, each consisting.