The effect of biochanin a and formononetin administration on preventing renal damage in lipopolysaccharide-induced endotoxemia

In this study, we tried to determine the extent to which biochanin A (BCA) and formononetin (FMN), which are known to have antioxidant and anti-inflammatory effects, can prevent oxidative stress and inflammatory response in kidney tissue against sepsis induced experimentally by lipopolysaccharide (LPS) induction. In our study, 42 female Wistar Albino rats (180-220 g), at least 8 weeks old, obtained from Van Yüzüncü Yıl University Experimental Application and Research Centre Directorate were used. The rats were randomly selected and divided into 6 groups with 7 animals in each group. These groups were respectively; control group, LPS group, LPS+BCA25 group, LPS+BCA50 group, LPS+FMN25 group and LPS+FMN50 group. No treatment was applied to the control group and they were sacrificed at the end of the study. The LPS group was administered 5 mg/kg LPS (i.p.) prepared by dissolving in 0.9% NaCl on the 7th day of the study and sacrificed at the end of the study. LPS+BCA and LPS+FMN groups were administered 25 and 50 mg/kg Biochanin A and Formononetin orally every day throughout the study, respectively. In addition, animals in this group were administered 5 mg/kg LPS (i.p.) prepared by dissolving in 0.9% NaCl on the 7th day of the study and sacrificed at the end of the study. At the end of the study, rats were anaesthetised with 10 mg/kg/i.p. xylazine and 50 mg/kg/i.p. ketamine, blood was collected by cardiac punction and kidney tissues were removed. The blood samples were centrifuged immediately and serum was obtained. In serum samples, urea, creatinine and blood urea nitrogen levels were determined by using a commercial kit in an autoanalyser. In kidney tissues, malondialdehyde (MDA) level was determined to determine lipid peroxidation, protein carbonyl level to determine protein oxidation, 8-hydroxy deoxy guanosine (8-OH dG) level to determine DNA oxidation, Superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px) and glutathione reductase (GSH-Rx) activities and reduced glutathione (GSH) levels were determined to determine the antioxidant status, and nitric oxide (NOx) and prostaglandin E2 (PGE2) levels were determined to observe the inflammatory response. Statistical analysis of the study was performed using SPSS v16.0 statistical programme. The data obtained were expressed as mean±standard deviation and data with a p value of 0.05 or less were considered statistically significant. One-way analysis of variance (ANOVA) and Tukey test were used in the evaluation of biochemical parameters. In LPS-induced sepsis, renal function tests were found to be impaired, lipid peroxidation, protein and DNA oxidation and inflammatory response in renal tissue were increased, and antioxidant defence system was weakened (p<0.05). Biochanin A and formononetin prevented oxidative stress and inflammation in kidney tissue (p<0.05). This study demonstrates that high doses of biochanin A and formononetin, which have proven antioxidant, anticancer, antiproliferative, antimetastatic, antiviral and antimicrobial effects, can be used prophylactically in LPS-induced sepsis, have antiinflammatory and antioxidant effects and prevent nephrotoxicity. Key Words: Lipopolysaccharide, Sepsis, Antioxidant, Anti-inflammatory, Isoflavones