Mk 2000

Systolic blood pressure (SBP) was measured by using the noninvasive tail-cuff plethysmography method and was recorded by using an automatic sphygmomanometer (MK2000; Muromachi Kikai, Tokyo, Japan). The systolic blood pressure (SBP) was measured at Weeks 4, 8, 12, and 16. The blood pressure of 7 mice per group was measured. At least five measurements were obtained at every session, and the mean of the five values within 5 mmHg was taken as the SBP level. Values are presented as the mean ± SEM of five measurements.

The systolic BP, diastolic BP, and heart rate were measured with a non-invasive blood pressure monitor tail-cuff method for rats (MK-2000, Muromachi Kikai, Tokyo, Japan) [39 (link)]. The BP and heart rate were monitored at 10 weeks of age and at 16 weeks prior to sacrifice. An echocardiogram was performed using the Vivid I ultrasound cardiovascular system (GE Healthcare, Haifa, Israel) at 10 weeks of age and at 16 weeks prior to sacrifice. The following cardiac structures were measured using M-Mode tracing of the LV: LVEDd, LVESd, EDV, ESV, EF, and FS values, as described previously [40 (link)].

Mice were anesthetized intraperitoneally with medetomidine hydrochloride (0.3 mg/kg body weight; Orion Corporation, Espoo, Finland), 4 mg/kg midazolam (4 mg/kg body weight; Sandoz, Tokyo, Japan), and butorphanol tartrate (5 mg/kg body weight; Meiji Seika, Tokyo, Japan). Experimental unilateral ureteral ligation resulting in UUO was performed (on day 0) in 8-week-old-male mice by ligation of the left ureter of each mouse at the ureteropelvic junction. Experiments were conducted in UUO kidneys (WT, n = 6 and Arg2 KO, n = 6). In Spd treatment experiments, five WT mice and five Arg2 KO mice were used. The mice were intraperitoneally administered with Spd (10 mg/kg, S0266; Sigma-Aldrich, St. Louis, MO, USA), or phosphate-buffered saline (Nacalai Tesque, Kyoto, Japan) once a day from day 0 for 2 weeks as reported previously^{47 (link)}. The body temperature of the mice was maintained at 37 °C during the whole procedure. Body weight was measured using an FX-3000 balance (A&D Company, Tokyo, Japan), and tail-cuff pressure was measured in a conscious state using a blood pressure monitor (MK-2000; Muromachi Kikai, Tokyo, Japan) before the operation and euthanasia.