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. 2020 Mar 1;318(3):F754-F762.
doi: 10.1152/ajprenal.00566.2019. Epub 2020 Feb 10.

Urethral dysfunction in a rat model of chemically induced prostatic inflammation: potential involvement of the MRP5 pump

Eduardo C Alexandre  1   2 , Nailong Cao  1 , Shinsuke Mizoguchi  1 , Tetsuichi Saito  1 , Masahiro Kurobe  1 , Daisuke Gotoh  1 , Meri Okorie  1 , Taro Igarashi  1 , Edson Antunes  2 , Naoki Yoshimura  1
Affiliations

Urethral dysfunction in a rat model of chemically induced prostatic inflammation: potential involvement of the MRP5 pump

Eduardo C Alexandre et al. Am J Physiol Renal Physiol. .

Abstract

Prostate inflammation (PI) is a clinical condition associated with infection and/or inflammation of the prostate. It is a common disease frequently associated to lower urinary tract (LUT) symptoms. The urethra is an understudied structure in the LUT and plays a fundamental role in the urinary cycle. Here, we proposed to evaluate the effect of PI on the urethra tissue. Male Sprague-Dawley rats were used, and PI was induced by formalin injection into the ventral lobes of the prostate. The pelvic urethra at the prostatic level was harvested for histological analysis, contraction (electrical field stimulation and phenylephrine), and relaxation (sodium nitroprusside/MK-571) experiments. Various gene targets [cytochrome c oxidase subunit 2, transforming growth factor-β1, interleukin-1β, hypoxia-inducible factor-1α, α1A-adrenoceptor, inositol 1,4,5-trisphosphate receptor type 1, voltage-gated Ca2+ channel subunit-α1D, neuronal nitric oxide synthase, soluble guanylyl cyclase, phosphodiesterase 5A, protein kinase CGMP-dependent 1, and multidrug resistance-associated protein 5 (MRP5; ATP-binding cassette subfamily C member 5)] were quantified, and cGMP levels were measured. No histological changes were detected, and functional assays revealed decreased contraction and increased relaxation of urethras from the PI group. The addition of MK-571 to functional assays increased urethral relaxation. Genes associated with inflammation were upregulated in urethras from the PI group, such as cytochrome oxidase c subunit 2, transforming growth factor-β1, interleukin-1β, and hypoxia-inducible factor-1α. We also found increased expression of L-type Ca2+ channels and the neuronal nitric oxide synthase enzyme and decreased expression of the MRP5 pump. Finally, cGMP production was enhanced in urethral tissue of PI animals. The results indicate that PI is associated with proinflammatory gene expression in the urethra without histologically evident inflammation and that PI produces a dysfunctional urethra and MRP5 pump downregulation, which results in cGMP accumulation inside the cell. These findings would help to better understand LUT dysfunctions associated with PI and the role of MRP pumps in the control of LUT function.

Keywords: cGMP; electrical field stimulation; multidrug resistance-associated protein 5; phenylephrine; prostatitis.

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Conflict of interest statement

No conflicts of interest, financial or otherwise, are declared by the author(s).

Figures

Fig. 1.
Fig. 1.
Sham (A and C) and prostate inflammation (PI; B and D) urethral transverse sections (8 µm) stained with hematoxylin and eosin. Images were captured using ×10 eyepieces with ×4 and ×10 objectives (n = 4−5).
Fig. 2.
Fig. 2.
Contraction responses to electrical field stimulation (1–32 Hz; A) and phenylephrine (PE; C) with representative original traces (B and D, respectively) in the urethra from sham and prostate inflammation (PI) groups. Data are presented as means ± SE; n = 6–7. *P < 0.05 compared with the sham group (Student’s t test).
Fig. 3.
Fig. 3.
Sodium nitroprusside (SNP)-induced relaxation (1, 10, and 100 μM) in rat urethras from the sham and prostate inflammation (PI) groups in the absence or presence of MK-571 (20 μM; A). Relaxations were calculated relative to the maximal changes from the contraction produced by phenylephrine (PE; 10 μM) in each urethral ring, which was taken as 100%. B: representative traces. Data are presented as means ± SE; n = 10. *P < 0.05 and **P < 0.01 compared with the sham group (one-way ANOVA followed by a Tukey test).
Fig. 4.
Fig. 4.
mRNA expression of cytochrome c oxidase subunit 2 (MTCO2, COX2; A), transforming growth factor-β1 (TGFB1, TGF-β1; B), interleukin-1β (IL1B, IL-1β; C), and hypoxia-inducible factor-1α (HIF1A, HIF-1α; D) in the urethra from sham and prostate inflammation (PI) groups. The mRNA expression level of each gene was normalized to the β-actin (ACTB) expression level. Data are presented as means ± SE; n = 7. *P < 0.05 and **P < 0.01 compared with the sham group (Student’s t test).
Fig. 5.
Fig. 5.
mRNA expressions of α1A-adrenoceptor (ADRA1A, α1A-AR; A), inositol 1,4,5-trisphosphate receptor type 1 (ITPR1, IP3R; B), voltage-gated Ca2+ channel subunit-α1D (CACNA1D, L-type Ca2+ channel; C), neuronal nitric oxide synthase (NOS2, nNOS; D), soluble guanylate cyclase (GUCY1B2, sGC; E), phosphodiesterase 5A (PDE5A, PDE5; F), protein kinase CGMP-dependent 1 (PRKG1, PKG; G), and ATP-binding cassette subfamily C member 5 [ABCC4, multidrug resistance-associated protein 5 (MRP5); H] in the urethra from sham and prostate inflammation (PI) groups. The mRNA expression level of each gene was normalized to the β-actin (ACTB) expression level. Data are presented as means ± SE; n = 6–7. *P < 0.05 compared with the sham group (Student’s t test).
Fig. 6.
Fig. 6.
cGMP content in urethral tissues from sham and prostate inflammation (PI) groups. Tissues were stimulated with sodium nitroprusside (SNP; 10 μM). Data are presented as means ± SE; n = 6. ***P < 0.001 compared with the sham group (Student’s t test).
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