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Proteomic analysis of urethral protein expression in an estrogen receptor α-deficient murine model of stress urinary incontinence

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Abstract

Purpose

The roles of estrogen receptor α (ERα) in stress urinary incontinence (SUI) remain elusive. This study was conducted to understand the molecular mechanism of ERα against SUI.

Methods

Wild-type (ERα+/+) and ACTB-cre ERα knockout (ERα−/−) female mice were generated. Urethral function and protein expression were measured. Leak point pressures (LPP) and maximum urethral closure pressure (MUCP) were assessed in mice under urethane anesthesia. After the measurements, the urethras were removed for proteomic analysis using the two-dimensional differential gel electrophoresis and liquid chromatography–mass spectrometry technology. Interaction between these ERα pathway proteins was further analyzed by using MetaCore. Lastly, Western blot and immunochemistry (IHC) were used to confirm the candidate protein expression levels and locations, respectively.

Results

Compared with the ERα+/+ group, the LPP and MUCP values of the ERα−/− group were significantly decreased. Additionally, we identified 11 differentially expressed proteins in the urethra of ERα−/− female mice; five proteins were down-regulated and six were up-regulated. The majority of the ERα knockout-modified proteins were involved in muscle development, contraction, and regulation, as well as immune response (amphoterin signaling and phagocytosis), proteolysis, and cell adhesion (platelet aggregation and integrin-mediated cell–matrix adhesion). IHC and Western blot confirmed the down-regulation of tropomyosin and up-regulation of myosin in urethra.

Conclusions

This is the first study to estimate protein expression changes in urethras from ERα−/− female mice. These changes could be related to the molecular mechanism of ERα in SUI.

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Acknowledgments

We are grateful for the technical assistance of Miss Chi-Hsiang Wei. This work was supported in part by Taiwan Ministry of Science and Technology (NSC 103-2911-I-002-303, NSC101-2632-B039-001-MY3, NSC101-2314-B-039-018, and NSC102-2320-B-039-025), China Medical University Hospital (DMR-104-058), National Health Research Institutes (NHRI-EX102-10241BI), CMU under the Aim for Top University Plan of the Taiwan Ministry of Education, and Taiwan Ministry of Health and Welfare Clinical Trial and Research Center of Excellence (MOHW103-TDU-B-212-113002 and MOHW104-TDU-B-212-113002).

Conflict of interest

The authors have no conflicts of interest to disclose.

Ethical standard

The procedures of the animal study received approval from the ethical committee of the Institutional Animal Care and Use Committee of China Medical University (Reference number: 101-201-N, Date: Feb. 3, 2012).

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    Authors and Affiliations

    1. Graduate Institute of Integrated Medicine, School of Chinese Medicine, College of Chinese Medicine, School of Pharmacy, College of Pharmacy, Department of Pharmacology, Department of Physical Therapy, Graduate Institute of Rehabilitation Science, Research Center for Chinese Medicine and Acupuncture, China Medical University, Taichung, 40402, Taiwan

      Yung-Hsiang Chen, Chao-Jung Chen, Yu-Ning Lin, Wen-Tsong Hsieh, Bor-Tsang Wu, Wen-Chi Chen, Kao-Sung Tsai, San-Yuan Wu & Huey-Yi Chen

    2. Departments of Medical Research, Urology, and Obstetrics and Gynecology, Proteomics Core Laboratory, Sex Hormone Research Center, Chinese Medicine Research and Development Center, Center for Molecular Medicine, China Medical University Hospital, Taichung, 40402, Taiwan

      Yung-Hsiang Chen, Chao-Jung Chen, Wen-Lung Ma, Wen-Chi Chen, Chawnshang Chang & Huey-Yi Chen

    3. Departments of Urology and Pathology, George H Whipple Laboratory for Cancer Research, Wilmot Cancer Center, University of Rochester Medical Center, Rochester, NY, 14642, USA

      Chawnshang Chang & Shuyuan Yeh

    4. Department of Psychology, College of Medical and Health Science, Asia University, Taichung, 41354, Taiwan

      Yung-Hsiang Chen

    5. Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan

      Yang-Chang Wu & San-Yuan Wu

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    1. Yung-Hsiang Chen
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    Correspondence to Huey-Yi Chen or Shuyuan Yeh.

    Additional information

    Y.-H. Chen and C.-J. Chen contributed equally to this study.

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    Chen, YH., Chen, CJ., Lin, YN. et al. Proteomic analysis of urethral protein expression in an estrogen receptor α-deficient murine model of stress urinary incontinence. World J Urol 33, 1635–1643 (2015). https://doi.org/10.1007/s00345-014-1474-3

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