The functions of the lower urinary tract, to storage and periodically release urine, are dependent on the activity of smooth and striated muscles in the bladder, urethra and external urethral sphincter. This activity is in turn controlled by neural circuits not only in the periphery, but also in the central nervous system (CNS). During urine storage, the outlet is closed and the bladder smooth muscle is quiescent by the neural control mechanism mainly organized in the spinal cord. When bladder volume reaches the micturition threshold, activation of a micturition center in the dorsolateral pons (the pontine micturition center) induces micturition through activation of sacral parasympathetic (pelvic) nerves. The brain rostral to the pons (diencephalon and cerebral cortex) is also involved in excitatory and inhibitory regulation of the micturition reflex. Various transmitters including dopamine, serotonin, norepenephrine, GABA, excitatory and inhibitory amino acids, opioids and acetylcholine are implicated in the modulation of the micturition reflex in the CNS. Therefore, injury or neurodegenerative diseases of the CNS as well as drugs can produce bladder and urethral dysfunctions such as urinary frequency, urgency and incontinence or inefficient bladder emptying.
Spinal cord injury (SCI) can lead to detrusor overactivity and detrusor-sphincter dyssynergia, which result in inefficient voiding and bladder wall tissue remodeling such as hypertrophy and fibrosis. However, no effective modality for controlling the bladder remodeling is available. In order to clarify whether an alpha1A/D-adrenoceptor (α1A/D-AR) antagonist, naftopidil, or a phosphodiesterase type 5 (PDE-5) inhibitor, tadalafil, prevents bladder wall remodeling after SCI, we examined the bladder and urethral activity as well as ischemic and fibrotic changes in the bladder using SCI rats with or without naftopidil or tadalafil treatment. Adult female Sprague-Dawley rats were divided into 4 groups; (1) normal (spinal cord intact), (2) vehicle SCI, (3) naftopidil SCI, and (4) tadalafil SCI groups. In SCI groups, rats underwent Th9-10 spinal cord transection followed by oral application of vehicle, naftopidil or tadalafil for 12 weeks. Bladder and urethral pressures, mRNA levels of fibrosis-related molecules and ischemia markers and the composition of bladder collagen and elastin were evaluated. Naftopidil treatment reduced the upregulation of mRNA levels of ischemia and fibrosis markers at the early phase of SCI, and ameliorated the decrease of bladder compliance and voiding efficiency, and the increase of collagen concentration in the bladder wall at the late phase of SCI. Tadalafil treatment reduced the upregulation of mRNA levels of fibrosis markers, the decrease of bladder compliance and the increase of collagen concentration at the late phase of SCI. These results suggest that PDE-5 inhibitors and α1A/D-AR antagonists treatments improved the bladder remodeling after SCI.
Stress urinary incontinence (SUI) is a common and bothersome problem among middle-aged women. However, there are few useful drugs for SUI. Urethral hypermobility and intrinsic sphincter deficiency are two main causes of SUI. Various animal models of SUI, such as vaginal distention, pudendal nerve injury, or ovariectomy, have been developed to study the pathophysiology of SUI. In addition, we have previously reported that cerebral infarction rats also induce SUI. Leak point pressure measurements are the most commonly used methods to evaluate the urethral dysfunction in SUI animal models. Originally, we have developed microtransducer-tipped catheter measurements of urethral activity during sneezing. Previous or our basic research has clarified potential strategies for pharmacotherapy of SUI in the central nervous system. Therapeutic targets include adrenergic and serotonergic (5-HT) receptors in the spinal cord, which stimulate pudendal nerve innervating the external urethral sphincter and/or sympathetic nerve innervating urethral smooth muscle. Activation of α1-adrenoceptors, 5-HT2C, or 5-HT7 receptors enhances the reflex at the spinal cord level whereas pre- or postsynaptic α2-adrenoceptors and/or 5-HT1A receptors inhibit the reflex. We have recently reported that stimulation of the spinal μ-opioid receptors by tramadol also enhances the reflex. Thus, we review the recent advances in basic SUI research and potential targets for pharmacotherapy of SUI in the central nervous system.
Psychological stress can induce not only frequent urination but also exacerbation of bladder dysfunctions. However, the brain pathophysiological mechanisms underlying stress-induced effects on the micturition reflex are still unknown. Bombesin (BB)-related peptides and BB receptors in the brain have been reported to mediate and integrate stress responses. We have found that centrally administered BB induced frequent urination in rats through brain BB1 and BB2 receptors, serotoninergic nervous system/5-HT7 receptors and corticotropin-releasing factor (CRF) type1 (CRF1) receptors. Interestingly, the BB-induced frequent urination was independent of the BB-induced activation of the sympatho-adrenomedullary outflow, a representative response to stress. Because the outflow is well known to regulate micturition, the finding was very surprising. These findings indicate that brain BB1, BB2, 5-HT7 and CRF1 receptors could be new therapeutic targets for bladder dysfunction exacerbated by stress exposure.
A molecular oxygen is essential to keep a physiological activity of each organ or a cell. There exists a heterogeneity in a level of oxygen concentration in each organ. In addition, tissue oxygen concentration fluctuates dynamically during physiological activities or in pathological processes. A decrease in tissue oxygen concentration, termed as hypoxia, significantly influences the function in each organ or cell. For example, a transcript level in each gene tends to be reduced under hypoxic condition. On the other hand, some of the gene expressions are increased significantly in hypoxia, which are termed as hypoxia responsive genes. A group of transcription factor, hypoxia inducible factor (HIF)-1α and HIF-2α play a critical role in the transactivation processes of hypoxia responsive genes. Recently, the molecular processes have been elucidated by which hypoxic environment activates HIF-1α or HIF-2α activity. A preclinical animal model revealed that HIF-α signal plays a critical role in inflammation or tissue remodeling. While HIF-1α and HIF-2α usually work synergistically in inducing their target gene expressions, macrophage HIF-1α and HIF-2α act antagonistically with regard to the synthesis of nitric oxide, a potent inflammatory mediator. This review summarizes the current understanding on the roles of HIF-α mediated hypoxic responses in inflammation or tissue remodeling.
Tubulointerstitial hypoxia negatively influences the balance between injury and repair, and serves as a final common pathway in chronic kidney disease (CKD). Studies on erythropoietin (EPO) transcription led to the identification of hypoxia inducible factors (HIFs) and their key regulators, prolyl hydroxylases (PHDs). Based on these, several small molecule PHD inhibitors are developed for the treatment of anemia in CKD, which are currently in phase II/III clinical trials. In addition to treating anemia, application of PHD inhibitors may have several potential implications; there is a promising view that activation of the HIF signaling might protect the ischemic kidney from injury. This is extensively tested in multiple acute kidney injury models, whereas knowledge is limited in the context of CKD. Some studies demonstrate the protective effects of ameliorating inflammation and reducing oxidative stress, whereas negative consequences of sustained HIF activation, such as renal fibrosis and aggravation of polycystic kidney disease, are also reported. Recent human clinical studies reported amelioration in glucose and lipid metabolism, which may be beneficial for the treatment of metabolic kidney disorders. Renal consequences of PHD inhibitors are likely determined by multiple systemic effects of sustained HIF activation and may thus differ depending on the clinical context and the pathological stages.
Tumor tissue environment is generally exposed to low oxygen, nutrition depletion and high interstitial pressure condition. These circumstances are caused by vascular hyper-permeability, irregular vascularization and immature vessels. The blood vessel is important tissue structures to deliver oxygen, nutrition and so on. An abnormal blood vessel formation is a common feature of tumor tissue that were characterized by hyper-permeability, irregular vascularization, immature vessels and intravasation. Therefore, tumor tissue is exposed to low oxygen nutrition depletion and low pH due to hypoperfusion and elevated interstitial pressure. These environments are one of the reasons for chemo- and radio-resistance. Previously, we reported that prolyl hydroxylase (PHD) inhibitor induced tumor blood vessel normalization and improved tumor microenvironment in tumor mouse model. However, effects of PHD inhibitor on tumor progression is controversial. Enhanced hypoxia inducible factors (HIFs) signaling in cancer cells act to promote cancer proliferation and metastases. On the other hand, increasing of HIFs signaling in immune cells may lead to activate inflammation and elicit anti-tumor effect. We describe our study how PHD inhibitor improved tumor microenvironment and focused on tumor infiltrate immune cells were phenotypic alteration after PHD inhibitor treatment in mouse model. Our results implied that PHD inhibitor was possibly beneficial for anti-cancer therapy.
Hypoxic responses are mainly regulated by heterodimeric transcription factor HIF, composed of unstable α-subunit (HIFα) and stable β-subunit (HIF1β/ARNT). Protein stability of HIFα depends on the hydroxylation status of its specific proline residue(s). Prolyl hydroxylation of HIFα is regulated by iron- and 2-oxoglutarate (2-OG)-dependent dioxygenase PHDs, whose enzyme activities are oxygen-dependent. Hence, PHDs act as an oxygen sensor, and inhibiting PHDs can activate the hypoxic response regardless of the normoxic environment. Small compounds that inhibit PHDs have been developed as the therapeutics for renal anemia. Here we also introduce the medical application of the PHD-inhibitors other than the renal anemia treatment. Finally, it is a great pleasure to announce here that the Nobel Prize in Physiology or Medicine 2019 was awarded to William G. Kaelin Jr, Sir Peter J. Ratcliffe, and Gregg L. Semenza, who have been studying how cells sense and adapt to oxygen availability over the years.
A computer simulation application on pharmacokinetics, which we developed using a software, named “Stella®”, has been successfully used for the virtual training of pharmacokinetics at multiple medical schools. The training course using Stella® has encouraged the medical students to optimize drug administration for individual patients on the computers. Importantly, the virtual training is free of any concern on human and animal ethics. The simulation application has been freely provided for medical schools without any restrictions and charge. For many years, it has been under constant version-upgrade in response to updates of the operating systems (OS) of personal computers or the software. Very recently, major updates of the OS and the software, and the emergence of tablet- and smartphones-type computers have been prompting us to perform a major revision of the simulation application. Here, we introduce the new version of the “web-based” simulation application that is available through any device including personal computers, tablets, and smartphones irrespective of the OSs (Microsoft Windows and Macintosh, Android, and iOS), without any extra charge unless the modification is required. We believe that the new-version of web-based simulation application will be useful not only for medical, nursing and pharmacy students, but also for medical workers who need to simulate drug pharmacokinetics on the computers before they administer drugs to the patients.