Inhibition of neurogenic and thromboxane A(2)-induced human prostate smooth muscle contraction by the integrin alpha 2 beta 1 inhibitor BTT-3033 and the integrin-linked kinase inhibitor Cpd22

Li, Bingsheng; Wang, Xiaolong; Wang, Ruixiao; Rutz, Beata; Ciotkowska, Anna; Gratzke, Christian; Herlemann, Annika; Spek, Annabel; Tamalunas, Alexander; Waidelich, Raphaela; Stief, Christian G. und Hennenberg, Martin (2020): Inhibition of neurogenic and thromboxane A(2)-induced human prostate smooth muscle contraction by the integrin alpha 2 beta 1 inhibitor BTT-3033 and the integrin-linked kinase inhibitor Cpd22. In: Prostate, Bd. 80, Nr. 11: S. 831-849

Abstract

Introduction: Prostate smooth muscle contraction is critical for etiology and treatment of lower urinary tract symptoms in benign prostatic hyperplasia (BPH). Integrins connect the cytoskeleton to membranes and cells to extracellular matrix, what is essential for force generation in smooth muscle contraction. Integrins are composed of different subunits and may cooperate with integrin-linked kinase (ILK). Here, we examined effects of inhibitors for different integrin heterodimers and ILK on contraction of human prostate tissues. Methods Prostate tissues were obtained from radical prostatectomy. Integrins and ILK were detected by Western blot, real-time polymerase chain reaction (RT-PCR), and double fluorescence staining. Smooth muscle contractions of prostate strips were studied in an organ bath. Contractions were compared after application of solvent (controls), the ILK inhibitor Cpd22 (N-methyl-3-(1-(4-(piperazin-1-yl)phenyl)-5-(4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)-1H-pyrazol-3-yl)propanamide), the integrin alpha 2 beta 1 inhibitor BTT-3033 (1-(4-fluorophenyl)-N-methyl-N-[4[[(phenylamino)carbonyl]amino]phenyl]-1H-pyrazole-4-sulfonamide), or the integrin alpha 4 beta 1/alpha 9 beta 1 inhibitor BOP (N-(benzenesulfonyl)- l-prolyl- l-O-(1-pyrrolidinylcarbonyl)tyrosine sodium salt). Results: Western blot analyses of prostate tissues using antibodies raised against integrins alpha 2b, alpha 4, alpha 9, beta 1, and ILK revealed bands matching the expected sizes of corresponding antigens. Expression of integrins and ILK was confirmed by RT-PCR. Individual variations of expression levels occurred independently from divergent degree of BPH, reflected by different contents of prostate-specific antigen. Double fluorescence staining of prostate sections using antibodies raised against integrins alpha 2 and beta 1, or against ILK resulted in immunoreactivity colocalizing with calponin, suggesting localization in prostate smooth muscle cells. Electric field stimulation (EFS) induced frequency-dependent contractions, which were inhibited by Cpd22 (3 mu M) and BTT-3033 (1 mu M) (inhibition around 37% by Cpd22 and 46% by BTT-3033 at 32 Hz). The thromboxane A(2) analog U46619-induced concentration-dependent contractions, which were inhibited by Cpd22 and BTT-3033 (around 67% by Cpd22 and 39% by BTT-3033 at 30 mu M U46619). Endothelin-1 induced concentration-dependent contractions, which were not affected by Cpd22 or BTT-3033. Noradrenaline and the alpha(1)-adrenergic agonists methoxamine and phenylephrine-induced concentration-dependent contractions, which were not or very slightly inhibited by Cpd22 and BTT-3033. BOP did not change EFS- or agonist-induced contraction. Conclusions: Integrin alpha 2 beta 1 and ILK inhibitors inhibit neurogenic and thromboxane A(2)-induced prostate smooth muscle contraction in human BPH. A role for these targets for prostate smooth muscle contraction may appear possible.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Medizin
• Themengebiete: 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
• ISSN: 0270-4137
• Sprache: Englisch