4.4. Conscious Cystometry

Cystometric studies were carried out after 17 days after the surgical procedures (i.e., 3 days after the last dose of the asiatic acid), as we described before [14,37]. Briefly, the bladder catheter was linked through a three-way stopcock to a pressure transducer (FT03; Grass Instruments) placed at the level of the bladder and to a microinjection pump (CMA 100; Microject, Solna, Sweden) for documenting intravesical pressure and for administrating physiological saline into the bladder. The bladder was physiologically filled with saline at a constant rate of 0.05 mL/min, i.e., 3 mL/h at room temperature (22 °C) to obtain repetitive voiding based on our previous studies [14]. The analog signal from the transducer was amplified and digitized using the Polyview system (Grass Instruments, Herlev, Denmark). Micturition amounts were measured using a fluid collector attached to a force displacement transducer (FT03C; Grass Instruments). Both transducers were connected to a polygraph (7 DAG; Grass Instruments). Cystometry profiles and micturition volumes were recorded continuously on a Grass polygraph (Model 7E; Grass Instruments). The values measured in each rat represent the average of five bladder micturition cycles after obtaining repetitive voiding. The mean calculations from all animals in each condition were averaged to create pooled data for each condition. All procedures were performed by a person blinded to the treatments.