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      Combination phosphodiesterase type 4 inhibitor and phosphodiesterase type 5 inhibitor treatment reduces non-voiding contraction in a rat model of overactive bladder

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          Abstract

          Introduction

          Current treatments for overactive bladder (OAB) are often discontinued due to side effects or lack of efficacy. The goal of this study was to determine if combining a phosphodiesterase type 4 inhibitor (PDE4i); with a type 5 inhibitor (PDE5i); would have a beneficial effect on OAB symptoms and if a reduced dose of PDE4i in combination with PDE5i could also provide a beneficial effect in OAB. We hypothesized that PDE5i and PDE4i combination treatment could be utilized to reduce non-voiding contractions and smooth muscle disruption in a rat model of OAB.

          Methods

          Fifty-eight age-matched Sprague-Dawley rats underwent PBOO and daily gavage with PDE4i alone (roflumilast; 1mg/kg), PDE5i alone (tadalafil;10mg/kg), high dose combination (PDE4i 1mg/kg, PDE5i 10mg/kg), low dose combination (PDE4i 0.2mg/kg, PDE5i 10mg/kg), or vehicle for 28 days. Fourteen animals underwent sham PBOO with vehicle. Rats underwent conscious and anesthetized cystometry 28 days after PBOO and were euthanized for qualitative bladder histology. One-way ANOVA on ranks with a Dunn’s post hoc test was used to indicate statistically significant differences between groups (p<0.05).

          Results

          Bladder & urethral weight was significantly increased after PBOO with vehicle, PDE4i alone, and PDE5i alone, but not with either combination treatment. Frequency of non-voiding contractions during both conscious and anesthetized cystometry increased significantly after PBOO with vehicle, but not after PDE4i or high dose combination treatments compared to sham PBOO. Threshold pressure for voiding was significantly decreased with high dose combination compared to vehicle. PBOO treated with PDE4i alone or high dose combination showed less bladder smooth muscle fibrosis than vehicle, PDE5i alone, or low dose combination treatments.

          Conclusion

          A PDE4i and PDE5i combination treatment has potential benefit in reducing OAB symptoms, but future research is needed.

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          Most cited references35

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          The standardisation of terminology in lower urinary tract function: report from the standardisation sub-committee of the International Continence Society.

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            Error bars in experimental biology

            Error bars commonly appear in figures in publications, but experimental biologists are often unsure how they should be used and interpreted. In this article we illustrate some basic features of error bars and explain how they can help communicate data and assist correct interpretation. Error bars may show confidence intervals, standard errors, standard deviations, or other quantities. Different types of error bars give quite different information, and so figure legends must make clear what error bars represent. We suggest eight simple rules to assist with effective use and interpretation of error bars.
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              Cyclic nucleotide-dependent relaxation pathways in vascular smooth muscle.

              Vascular smooth muscle tone is controlled by a balance between the cellular signaling pathways that mediate the generation of force (vasoconstriction) and release of force (vasodilation). The initiation of force is associated with increases in intracellular calcium concentrations, activation of myosin light-chain kinase, increases in the phosphorylation of the regulatory myosin light chains, and actin-myosin crossbridge cycling. There are, however, several signaling pathways modulating Ca(2+) mobilization and Ca(2+) sensitivity of the contractile machinery that secondarily regulate the contractile response of vascular smooth muscle to receptor agonists. Among these regulatory mechanisms involved in the physiological regulation of vascular tone are the cyclic nucleotides (cAMP and cGMP), which are considered the main messengers that mediate vasodilation under physiological conditions. At least four distinct mechanisms are currently thought to be involved in the vasodilator effect of cyclic nucleotides and their dependent protein kinases: (1) the decrease in cytosolic calcium concentration ([Ca(2+)]c), (2) the hyperpolarization of the smooth muscle cell membrane potential, (3) the reduction in the sensitivity of the contractile machinery by decreasing the [Ca(2+)]c sensitivity of myosin light-chain phosphorylation, and (4) the reduction in the sensitivity of the contractile machinery by uncoupling contraction from myosin light-chain phosphorylation. This review focuses on each of these mechanisms involved in cyclic nucleotide-dependent relaxation of vascular smooth muscle under physiological conditions.
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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Project administrationRole: ValidationRole: Writing – original draftRole: Writing – review & editing
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: Project administrationRole: Writing – original draftRole: Writing – review & editing
                Role: Data curationRole: Formal analysis
                Role: InvestigationRole: MethodologyRole: Project administrationRole: Writing – original draftRole: Writing – review & editing
                Role: MethodologyRole: Project administrationRole: Writing – original draftRole: Writing – review & editing
                Role: Project administrationRole: ResourcesRole: Writing – review & editing
                Role: Project administrationRole: ResourcesRole: Writing – review & editing
                Role: Formal analysisRole: MethodologyRole: Project administrationRole: SupervisionRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                28 August 2019
                2019
                : 14
                : 8
                : e0220788
                Affiliations
                [1 ] Department of Biomedical Engineering, Lerner Research Institute, the Cleveland Clinic, Cleveland, Ohio, United States of America
                [2 ] Advanced Platform Technology Center, Louis Stokes Cleveland Veterans Affairs Medical Center, Cleveland, Ohio, United States of America
                [3 ] Department of Biology, University of Akron, Akron, Ohio, United States of America
                [4 ] Department of Urology, Changhai Hospital, Naval Medical University, Shanghai, People’s Republic of China
                [5 ] Department of Obstetrics and Gynecology, the Cleveland Clinic, Cleveland, Ohio, United States of America
                [6 ] Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana, United States of America
                [7 ] Glickman Urological and Kidney Institute, the Cleveland Clinic, Cleveland, Ohio, United States of America
                Max Delbruck Centrum fur Molekulare Medizin Berlin Buch, GERMANY
                Author notes

                Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: LLP is an employee of Eli Lilly. GGD is an employee of Eli Lilly. MSD was principal investigator of a sponsored research agreement with Eli Lilly at the Cleveland Clinic. The funder provided support in the form of salaries for the authors LLP and GGD, but did not have any additional role in the data collection and analysis, decision to publish, or preparation of the manuscript. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

                Author information
                http://orcid.org/0000-0003-4743-9283
                Article
                PONE-D-19-06317
                10.1371/journal.pone.0220788
                6713339
                31461445
                bd337f61-25cf-470e-a1f7-b31c26e7fd5c

                This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

                History
                : 4 March 2019
                : 23 July 2019
                Page count
                Figures: 5, Tables: 2, Pages: 15
                Funding
                This work was done with a sponsored research agreement to the Cleveland Clinic from Eli Lilly (P.I. MSD). The funder provided support in the form of salaries for authors LLP and GGD, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section. Eli Lilly employees participated in study design and preparation of manuscript, but final decisions were made by Dr. Damaser and her group at the Cleveland Clinic. Data collection and analysis was done solely by Dr. Damaser and her group. Additional support from the Cleveland Clinic and the U.S. Department of Veterans Affairs was provided.
                Categories
                Research Article
                Biology and Life Sciences
                Anatomy
                Renal System
                Bladder
                Medicine and Health Sciences
                Anatomy
                Renal System
                Bladder
                Biology and Life Sciences
                Bioengineering
                Biotechnology
                Medical Devices and Equipment
                Catheters
                Engineering and Technology
                Bioengineering
                Biotechnology
                Medical Devices and Equipment
                Catheters
                Medicine and Health Sciences
                Medical Devices and Equipment
                Catheters
                Biology and Life Sciences
                Anatomy
                Musculoskeletal System
                Muscles
                Smooth Muscles
                Medicine and Health Sciences
                Anatomy
                Musculoskeletal System
                Muscles
                Smooth Muscles
                Biology and Life Sciences
                Developmental Biology
                Fibrosis
                Biology and Life Sciences
                Biochemistry
                Enzymology
                Enzymes
                Phosphodiesterases
                Biology and Life Sciences
                Biochemistry
                Proteins
                Enzymes
                Phosphodiesterases
                Medicine and Health Sciences
                Euthanasia
                Medicine and Health Sciences
                Urology
                Bladder and Ureteric Disorders
                Cystitis
                Biology and Life Sciences
                Bioengineering
                Biotechnology
                Medical Devices and Equipment
                Medical Implants
                Engineering and Technology
                Bioengineering
                Biotechnology
                Medical Devices and Equipment
                Medical Implants
                Medicine and Health Sciences
                Medical Devices and Equipment
                Medical Implants
                Custom metadata
                Data are available at Dryad ( https://doi.org/10.5061/dryad.q7n55m2).

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