Combination of Wu Lin San and Shan Zha ameliorates substance P-induced hyperactive bladder via the inhibition of neutrophil NADPH oxidase activity

Substance P (SP) via neurokinin type 1 receptor activates leukocytes to produce burst release of reactive oxygen species (ROS) and increases leukocytes adhesion to the vessels in the inflamed bladder. Activation of neutrophil nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity may contribute to the neutrophil ROS production. We explored the therapeutic effect of traditional Chinese formula for urinary dysfunction, Wu Lin San (WLS), and a modified formula WLS plus Shan Zha (WLSSZ) on SP-induced bladder hyperactivity. We evaluated WLS, Shan Zha, and WLSSZ effect on neutrophils NADPH oxidase activity in SP-stimulated neutrophils in vitro, and isovolumetric cystometrogram and ROS activity in vivo in anesthetized rat bladder with SP stimulation. Our results showed that WLS, Shan Zha, and WLSSZ inhibited SP-induced NADPH oxidase activity in an order WLSSZ > Shan Zha > WLS. Exogenous SP enhanced systemic vasodilation, bladder hyperactivity and bladder ROS. One week of oral administration of WLS or WLSSZ significantly reduced SP-induced bladder ROS amount and leukocyte accumulation and ameliorated the hyperactive bladder response. The therapeutic action was better in WLSSZ than in WLS. Our results indicate that a modified formula Wu Lin San plus Shan Zha can potentially ameliorate SP-induced neurogenic inflammation possibly via the inhibition of leukocyte NADPH oxidase activity.