Bladder Dysfunction in an Obese Zucker Rat: The Role of TRPA1 Channels, Oxidative Stress, and Hydrogen Sulfide

Figure 5

Downregulation of the CSE/H₂S pathway in the OZR bladder. CBS and CSE expression in representative detrusor muscle transverse sections from a total of 5 lean Zucker rat (LZR) and obese Zucker rat (OZR) bladders (a–h). An intense CBS immunostaining in the smooth muscle (SM) and urothelium (URO) of bladders from the LZR and OZR was shown (a–d), whereas CSE staining was weaker (e–h). Both cytoplasm of the smooth muscle and urothelial cells of the bladder from the obese group (f and h) showed lower CSE staining versus that observed in the LZR (e and g). CBS immunoreactivity was similar in the LZR and OZR (a–d). Western blot of membranes from the LZR and OZR showing 63 kDa and 45 kDa major bands, which corresponded to the expected molecular weight for CBS and CSE, respectively, indicating a reduced CSE protein expression in the OZR bladder (j) (, vs. the LZR value, unpaired Student’s -test). CBS protein expression was similar in bladders from the LZR and OZR (i). The level of H₂S generated in the LZR and OZR bladders, in the absence (control) or presence of aminooxyacetic acid (AOAA, 1 mM) and DL-propargylglycine (PPG, 1 mM), inhibitors of CBS and CSE, respectively. Endogenous H₂S production was significantly reduced in the OZR. Bars represent of the 3 LZR and OZR. vs. LZR control values. LZR control vs. LZR AOAA. LZR control vs. LZR PPG (ANOVA followed by the Bonferroni method) (k).