Characterizing relaxin receptor expression and exploring relaxin’s effect on tissue remodeling/fibrosis in the human bladder
Abstract Background Relaxin is an endogenous protein that has been shown to have antifibrotic properties in various organ systems. There has been no characterization of relaxin’s role in the human bladder. Our objective was to characterize relaxin receptor expression in the human bladder and assess...
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| Format: | Article |
| Language: | English |
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BMC
2020-04-01
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| Series: | BMC Urology |
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| Online Access: | http://link.springer.com/article/10.1186/s12894-020-00607-4 |
| _version_ | 1819173145959989248 |
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| author | Edward C. Diaz Mason Briggs Yan Wen Guobing Zhuang Shannon L. Wallace Amy D. Dobberfuhl Chia-Sui Kao Bertha C. Chen |
| author_facet | Edward C. Diaz Mason Briggs Yan Wen Guobing Zhuang Shannon L. Wallace Amy D. Dobberfuhl Chia-Sui Kao Bertha C. Chen |
| author_sort | Edward C. Diaz |
| collection | DOAJ |
| description | Abstract Background Relaxin is an endogenous protein that has been shown to have antifibrotic properties in various organ systems. There has been no characterization of relaxin’s role in the human bladder. Our objective was to characterize relaxin receptor expression in the human bladder and assess relaxin’s effect on tissue remodeling/fibrosis pathways in bladder smooth muscle cells. Methods Relaxin family peptide receptor 1 (RXFP1) and RXFP2 expression was assessed using quantitative reverse transcriptase-PCR (qRT-PCR) and immunohistochemistry (IHC) on primary bladder tissue. Primary human smooth muscle bladder cells were cultured and stimulated with various concentrations of relaxin. Western blot, qRTPCR, ELISA, and zymogram assays were used to analyze fibrosis/tissue remodeling pathway proteins. Results There was universal mRNA transcript detection and protein expression of relaxin receptors in primary bladder specimens. Immunohistochemistry demonstrated RXFP1 and RXFP2 localizing to both urothelial and smooth muscle cell layers of the bladder. 24 h of in vitro relaxin stimulation did not affect mRNA expression of selected proteins in human bladder smooth muscle cells. However, 48 h of in vitro relaxin stimulation resulted in upregulation of active (p = 0.004) and latent (p = 0.027) MMP-2 in cell lysate, and upregulation of active MMP-2 in supernatant (p = 0.04). There was a dose dependent relationship with increasing expression of MMP-2 with increasing relaxin concentration. Relaxin stimulation resulted in decreased levels of active and total TGF-β1 in supernatant and extracellular matrix (p < 0.005 with 100 ng/mL relaxin stimulation). Conclusions In the human bladder, relaxin receptors are expressed at the dome and trigone and localize to the urothelium and smooth muscle cell layers. Stimulation of human bladder SMCs with relaxin in vitro affects expression of MMP-2 and TGF-β1. |
| first_indexed | 2024-12-22T20:18:26Z |
| format | Article |
| id | doaj.art-359d46a5d68c4a9cb629c53e959e949c |
| institution | Directory Open Access Journal |
| issn | 1471-2490 |
| language | English |
| last_indexed | 2024-12-22T20:18:26Z |
| publishDate | 2020-04-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Urology |
| spelling | doaj.art-359d46a5d68c4a9cb629c53e959e949c2022-12-21T18:13:55ZengBMCBMC Urology1471-24902020-04-012011910.1186/s12894-020-00607-4Characterizing relaxin receptor expression and exploring relaxin’s effect on tissue remodeling/fibrosis in the human bladderEdward C. Diaz0Mason Briggs1Yan Wen2Guobing Zhuang3Shannon L. Wallace4Amy D. Dobberfuhl5Chia-Sui Kao6Bertha C. Chen7Department of Urology, Stanford University Medical CenterDepartment of Urology, Stanford University Medical CenterDepartment of Obstetrics and Gynecology, Stanford University Medical CenterDepartment of Obstetrics and Gynecology, Stanford University Medical CenterDepartment of Obstetrics and Gynecology, Stanford University Medical CenterDepartment of Urology, Stanford University Medical CenterDepartment of Pathology, Stanford University Medical CenterDepartment of Obstetrics and Gynecology, Stanford University Medical CenterAbstract Background Relaxin is an endogenous protein that has been shown to have antifibrotic properties in various organ systems. There has been no characterization of relaxin’s role in the human bladder. Our objective was to characterize relaxin receptor expression in the human bladder and assess relaxin’s effect on tissue remodeling/fibrosis pathways in bladder smooth muscle cells. Methods Relaxin family peptide receptor 1 (RXFP1) and RXFP2 expression was assessed using quantitative reverse transcriptase-PCR (qRT-PCR) and immunohistochemistry (IHC) on primary bladder tissue. Primary human smooth muscle bladder cells were cultured and stimulated with various concentrations of relaxin. Western blot, qRTPCR, ELISA, and zymogram assays were used to analyze fibrosis/tissue remodeling pathway proteins. Results There was universal mRNA transcript detection and protein expression of relaxin receptors in primary bladder specimens. Immunohistochemistry demonstrated RXFP1 and RXFP2 localizing to both urothelial and smooth muscle cell layers of the bladder. 24 h of in vitro relaxin stimulation did not affect mRNA expression of selected proteins in human bladder smooth muscle cells. However, 48 h of in vitro relaxin stimulation resulted in upregulation of active (p = 0.004) and latent (p = 0.027) MMP-2 in cell lysate, and upregulation of active MMP-2 in supernatant (p = 0.04). There was a dose dependent relationship with increasing expression of MMP-2 with increasing relaxin concentration. Relaxin stimulation resulted in decreased levels of active and total TGF-β1 in supernatant and extracellular matrix (p < 0.005 with 100 ng/mL relaxin stimulation). Conclusions In the human bladder, relaxin receptors are expressed at the dome and trigone and localize to the urothelium and smooth muscle cell layers. Stimulation of human bladder SMCs with relaxin in vitro affects expression of MMP-2 and TGF-β1.http://link.springer.com/article/10.1186/s12894-020-00607-4RelaxinFibrosisBladderRXFP1RXFP2 |
| spellingShingle | Edward C. Diaz Mason Briggs Yan Wen Guobing Zhuang Shannon L. Wallace Amy D. Dobberfuhl Chia-Sui Kao Bertha C. Chen Characterizing relaxin receptor expression and exploring relaxin’s effect on tissue remodeling/fibrosis in the human bladder BMC Urology Relaxin Fibrosis Bladder RXFP1 RXFP2 |
| title | Characterizing relaxin receptor expression and exploring relaxin’s effect on tissue remodeling/fibrosis in the human bladder |
| title_full | Characterizing relaxin receptor expression and exploring relaxin’s effect on tissue remodeling/fibrosis in the human bladder |
| title_fullStr | Characterizing relaxin receptor expression and exploring relaxin’s effect on tissue remodeling/fibrosis in the human bladder |
| title_full_unstemmed | Characterizing relaxin receptor expression and exploring relaxin’s effect on tissue remodeling/fibrosis in the human bladder |
| title_short | Characterizing relaxin receptor expression and exploring relaxin’s effect on tissue remodeling/fibrosis in the human bladder |
| title_sort | characterizing relaxin receptor expression and exploring relaxin s effect on tissue remodeling fibrosis in the human bladder |
| topic | Relaxin Fibrosis Bladder RXFP1 RXFP2 |
| url | http://link.springer.com/article/10.1186/s12894-020-00607-4 |
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