Mrl lpr mice

Eight-week-old female MRL/lpr mice were purchased from Jackson Laboratories (Bar Harbor, ME, USA). The mice were housed and maintained in the animal facilities of Tohoku University School of Medicine. Bz was purchased from Santa Cruz (Dallas, TX, USA) and prepared at a concentration of 10 mg/ml. Cyclophosphamide (Cyc) was purchased from Sigma-Aldrich (St. Louis, MO, USA) and prepared at a concentration of 5 mg/ml. We injected the mice subcutaneously with Bz (750 μg/kg body) twice weekly or intraperitoneally with Cyc (1 mg/body = 30–35 mg/kg) once in 2 weeks at the ages indicated. Control mice were injected subcutaneously or intraperitoneally with the same volume of phosphate buffered saline (PBS) instead of Bz and Cyc, respectively. The mice were euthanized by cervical dislocation at the specified ages, and samples were analyzed for immunological activities and to assess the effects and toxicities of drugs. The study protocols were reviewed by the Institutional Laboratory Animal Care and Use Committee of Tohoku University, and finally approved by the President of University.

Female C57BL/6 (B6) and MRL/lpr mice were purchased from the Jackson Laboratory (Bar Harbor, ME, USA) and maintained in a specific pathogen-free colony. Animal experiments were approved and conducted in accordance with University of Houston’s Institutional Animal Care regulations. Twenty-eight 4-month-old MRL/lpr mice were divided into a control group (n = 14) and a treatment group (n = 14) randomly for the in vivo studies. The B1R antagonist SSR240612 was purchased from Adooq Bioscience (Irvine, CA, USA). SSR240612 was dissolved in water containing dimethyl sulfoxide (DMSO) to make a final concentration of 1.5 mg/mL in 0.9% DMSO. Mice in the treatment group were administered 10 mg/kg per day SSR240612 by gavage every other day, whereas the mice in the control group received 10 mg/kg per day 0.9% DMSO by gavage every other day; 24-h urine was collected using metabolic cages from all mice. Blood and urine were collected at 0, 8, and 12 weeks after treatment to assess proteinuria, serum blood urea nitrogen (BUN), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). At 12 weeks after treatment, all mice were euthanized by using a CO₂ chamber and cervical dislocation.