Fig 2: TNF treatment on primary hypothalamic neurons. (A) B1R mRNA levels in cultured neurons, measured by real time PCR (n = 6 independent cultures/group). (B) ROS generation indicated by microplate DHE assay (n = 6–7/group). (C) Quantification and immunofluorescent images of B1R expression (n = 8–10/group). (D) ROS production and quantification of DHE staining (n = 7/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p < 0.05 compared to vehicle. † p < 0.05 compared to TNF.

Fig 3: H2O2 treatment on primary hypothalamic neurons. (A) B1R expression indicated by immunofluorescence and quantification (n = 6–10/group). (B) Generation of reactive oxygen species using DHE staining (n = 7/group). (C) DHE microplate assay revealing ROS generation (n = 6/group). Statistical significance: One-way ANOVA followed by Tukey’s multiple comparisons test. * p < 0.05 compared to vehicle. † p < 0.05 compared to H2O2.

Fig 4: Kinin B1R mediates the activation of orexin system and hypertension. Bradykinin metabolites or kinin B1 receptor (B1R) agonists activate the B1R in the brain, which in turn modulates the expression of both orexin system and arginine vasopressin (AVP). This dual activation and increase of orexin and AVP contribute to the development and sustainment of neurogenic hypertension.

Fig 5: B1R gene deletion abrogates DOCA-salt induced renal myofibroblast transition. (A) Representative immunofluorescence images showing expression of alpha-smooth muscle actin (α-SMA), fibronectin, and transforming growth factor-beta (TGF-β) in the kidney sections. Quantification data presented as mean fluorescence staining showing increased expression of α-SMA (B), fibronectin (C), and TGF-β (D) of DOCA-salt treated hypertensive mice compared with sham mice. These increases in fibrosis markers expression were blunted in B1R deficient mice with 3 weeks of DOCA-salt treatment (n = 6, Two-way ANOVA, *P < 0.01 vs. Sham, †P < 0.05 vs. WT + DOCA).