Figure 6. Mice with GPR35 (G-protein–coupled receptor 35) deletion showed resistance to blood pressure elevation and protection of endothelial function after deoxycorticosterone acetate (DOCA)-salt treatment.

A, The Gpr35 mRNA level in mouse aortic endothelial cells (MAECs) isolated from GPR35^WT mice received DOCA-salt or sham treatment. n=5 in sham group, n=7 in DOCA group. Mann-Whitney U test did not reach statistical significance. B, The systolic blood pressure (SBP) and diastolic blood pressure (DBP) at the end of the experiment (BP_{week 4}) in the animals receiving sham procedure, DOCA-salt procedure, or DOCA-salt with N^G-nitro-L-arginine methyl ester hydrochloride (L-NAME). The n numbers are shown underneath each group on x axis. The mean values of each group are shown underneath the dot plot. One-way ANOVA test was done, then the pairs of focused groups were tested with a 2-sample t test and P values were adjusted with Hommel method. The symbols denote significant difference found between the 2 groups in post hoc comparisons. *P<0.05 vs GPR35^WT-Sham, #P<0.05 vs GP35^KO-Sham, $P<0.05 vs GPR35^KO-DOCA. C, Change in SBP and DBP in male GPR35^KO and male GPR35^WT mice after sham, DOCA-salt, or DOCA-salt with L-NAME procedures (BP_{week 4}−BP_{week 0}). The n numbers are shown underneath each group on x axis. The mean values of each group are shown underneath the dot plot. One-way ANOVA test was done, and if significant, pairs of focused groups were tested with a 2-sample t test and P values were adjusted with Hommel method. The symbols denote significant difference found between the 2 groups in post hoc comparisons. *P<0.05 vs GPR35^WT-Sham between the 2 groups, #P<0.05 vs GP35^KO-Sham, $P<0.05 vs GPR35^KO-DOCA. D, Quantification of reactive oxygen species by dihydroethidium staining in aortas isolated from GPR35^WT and GPR35^KO mice with DOCA-salt or Sham treatment. The n numbers are shown underneath each group on x axis. Kruskal-Wallis tests were done, followed by Mann-Whitney U tests with Hommel adjustment. *P<0.05 between the 2 groups in post hoc comparisons. E, Representative images of aortas isolated from GPR35^WT or GPR35^KO mice received DOCA-salt or Sham treatment. Aortas were stained with dihydroethidium, DAPI, and EC marker CD31 or smooth muscle cell marker SM22. Scale bar=400 μm. In all the figures, horizontal lines show mean±SD. F, The proposed mechanism by which GPR35 participates in the regulation of endothelial function and BP.