Functional bowel disorders (FBD) have a major potential to degrade the standards of public life. Juniperus oxycedrus L. (J. oxycedrus) (Cupressaceae) has been described as a plant used in traditional medicine as an antidiarrheal medication. The present study is the first to obtain information on the antispasmodic and antidiarrheic effects of J. oxycedrus aqueous extract through in vitro and in vivo studies. An aqueous extract of J. oxycedrus (AEJO) was extracted by decoctioning air-dried aerial sections of the plant. Antispasmodic activity was tested in an isolated jejunum segment of rats exposed to cumulative doses of drogue extract. The antidiarrheic activity was tested using diarrhea caused by castor oil, a transit study of the small intestine, and castor oil-induced enteropooling assays in mice. In the jejunum of rats, the AEJO (0.1, 0.3 and 1 mg/ml) diminished the maximum tone induced by low K+ (25 mM), while it exhibited a weak inhibitory effect on high K+ (75 mM) with an IC50=0.49 ± 0.01 mg/ml and IC50=2.65 ± 0.16 mg/ml, respectively. In the contractions induced by CCh (10-6 M), AEJO diminished the maximum tone, similar to that induced by low K+ (25 mM). with an IC50=0.45 ± 0.02 mg/ml. The inhibitory effect of AEJO on low K+ induced contractions was significantly diminished in the presence of glibenclamide (GB) (0.3 µM) and 4-aminopyrimidine (4-AP) (100 µM), with IC50 values of 1.84 ± 0.09 mg/ml. and 1.63 ± 0.16 mg/ml, respectively). The demonstrated inhibitory effect was similar to that produced by a non-competitive antagonist acting on cholinergic receptors and calcium channels. In castor oil-induced diarrhea in mice, AEJO (100, 200, and 400 mg/kg) caused an extension of the latency time, a reduced defecation frequency, and a decrease in the amount of wet feces compared to the untreated group (distilled water). Moreover, it showed a significant anti-motility effect and reduced the amount of fluid accumulated in the intestinal lumen at all tested doses. These findings support the conventional use of Juniperus oxycedrus L. as a remedy for gastrointestinal diseases.

Macrophages are the originators of inflammatory compounds, phagocytic purifiers in their local environment, and wound healing protectors in oxidative environments. They are molded by the tissue milieu they inhabit, with gastrointestinal (GI) muscularis macrophages (MMs) being a prime example. MMs are located in the muscular layer of the GI tract and contribute to muscle repair and maintenance of GI motility. MMs are often in close proximity to the enteric nervous system, specifically near the enteric neurons and interstitial cells of Cajal (ICCs). Consequently, the anti-inflammatory function of MMs corresponds to the development and maintenance of neural networks in the GI tract. The capacity of MMs to shift from anti-inflammatory to proinflammatory states may contribute to the inflammatory aspects of various GI diseases and disorders such as diabetic gastroparesis or postoperative ileus, functional disorders such as irritable bowel syndrome, and organic diseases such as inflammatory bowel disease. We reviewed the current knowledge of MMs and their influence on neighboring cells due to their important role in the GI tract.

In contrast to the long-standing focus on the pathophysiology of skeletal muscles in thehunt for a cure for Duchenne muscular dystrophy (DMD), we opine that the malfunctioning ofdystrophin produced by vascular smooth muscle is a major contributor to the pathology ofthe illness. We believe that a biological response modifier glucan (BRMG), which has beenshown in clinical studies of DMD to boost the expression of vascular smooth muscledystrophin and provide anti-fibrotic and anti-inflammatory effects, may play a key role inreducing the pathogenesis of DMD. According to the evaluation of biomarkers, this BRMG,which is safe and side-effect-free, reduces the pathogenesis of DMD. We describe thepossible mechanisms of action by which this BRMG helps in alleviating the symptoms of DMDby targeting smooth muscle dystrophin, in addition to its advantages over othertherapeutic modalities, as well as how it can serve as a valuable adjunct to existingtherapies. We suggest that using BRMG adjuncts that target smooth muscle dystrophin wouldbe a potential therapeutic approach that prolongs the lifespan and extends the duration ofambulation from the onset of DMD. Further studies are needed to validate thishypothesis.

In the lower urinary tract, transient receptor potential (TRP) channels are primarilyinvolved in physiological function, especially in cellular sensors responding to chemicaland physical stimuli. Among TRP channels, TRP melastatin 8 (TRPM8) channels, responding tocold temperature and/or chemical agents, such as menthol or icilin, are mainly expressedin the nerve endings of the primary afferent neurons and in the cell bodies of dorsal rootganglia innervating the urinary bladder (via Aδ- and C-fibers); this suggests that TRPM8channels primarily contribute to bladder sensory (afferent) function. Storage symptoms ofoveractive bladder, benign prostatic hyperplasia, and interstitial cystitis are commonlyrelated to sensory function (bladder hypersensitivity); thus, TRPM8 channels may alsocontribute to the pathophysiology of bladder hypersensitivity. Indeed, it has beenreported in a pharmacological investigation using rodents that TRPM8 channels contributeto the pathophysiological bladder afferent hypersensitivity of mechanosensitive C-fibers.Similar findings have also been reported in humans. Therefore, a TRPM8 antagonist would bea promising therapeutic target for bladder hypersensitive disorders, including urinaryurgency or nociceptive pain. In this review article, the functional role of the TRPM8channel in the lower urinary tract and the potential of its antagonist for the treatmentof bladder disorders was described.

Background/Aims: Gastroprokinetic agents are used for patients withpostoperative ileus (POI), and the Japanese traditional herbal medicine daikenchuto (DKT)is one such agent used in the clinical setting. POI is caused by inflammation. DKT andrikkunshito have anti-inflammatory abilities in addition to their gastroprokineticeffects. The efficacy of Kampo formulations, including hangekobokuto (HKT), in patientswith POI has been reported recently. Several authors have described the efficacy ofhonokiol, the primary component of Magnoliae Cortex, in HKT in mousemodels of POI. We therefore analyzed the effect of HKT on POI model mice to determine thesimilarities in the mechanism of action between HKT and DKT. Methods: HKT was administered orally to each mouse before and afterintestinal manipulation was performed on the distal ileum. The gastrointestinal transitin vivo, leukocyte infiltration, and levels of inflammatory mediators,such as cytokines and chemokines, were analyzed. Results: HKT significantly inhibited the infiltration of neutrophils andmacrophages and led to the recovery of delayed intestinal transit. In addition, itsignificantly decreased inducible nitric oxide synthase (iNOS) as well as honokiol levels,suggesting anti-inflammatory activity. However, it did not inhibit the increase in levelsof interleukin (IL)-1beta and IL-6, which are related to iNOS induction. In contrast, HKTincreased levels of nerve growth factor (NGF) and suppressed those of nuclear factor-κB(NFκB), which are related to iNOS induction, suggesting the possibility of a neuronalanti-inflammatory mechanism. Conclusions: HKT exerted a POI-relieving effect similar to DKT in a murinePOI model, and findings suggest that it may exert its anti-inflammatory activity throughNGF.

Vascular smooth muscle cells (VSMCs) and endothelial cells (ECs) are major cell typesthat control vascular function, and hence dysfunction of these cells plays a key role inthe development and progression of vasculopathies. Abnormal vascular responsiveness tovasoactive substances including vasoconstrictors and vasodilators has been observed invarious arteries in diseases including diabetes, hypertension, chronic kidney diseases,and atherosclerosis. Several substances derived from ECs tightly control vascularfunction, such as endothelium-derived relaxing and contracting factors, and it is knownthat abnormal vascular signaling of these endothelium-derived substances is often observedin various diseases. Derangement of signaling in VSMCs and altered function influencevascular reactivity to vasoactive substances and tone, which are important determinants ofvascular resistance and blood pressure. However, understanding the molecular mechanismsunderlying abnormalities of vascular functions in pathological states is difficult becausemultiple substances interact in the development of these processes. Advanced glycation endproducts (AGEs), a heterogeneous group of bioactive compounds, are thought to contributeto vascular dysfunction, which in turn cause the development of several diseases includingdiabetes, hypertension, stroke, and atherosclerosis. A growing body of evidence suggeststhat AGEs could affect these cells and modulate vascular function. This study is focusedon the link between AGEs and functions of ECs and VSMCs, particularly the modulativeeffects of AGEs on vascular reactivities to vasoactive substances.

The characteristic mechanical activities of the smooth muscles found in all organs of thebody are highly variable and depend mainly on the spatial arrangement of the muscle cellsand the stroma: mass, orientation, relationships, links, constraints, which are deployedin various configurations. These structural features are examined here for theirmechanical relevance, in light and electron microscopic views of several muscles ofviscera and blood vessels, in a selection of mammalian species. Smooth muscles areincompressible and therefore maintain constant volume. They do not have available spaceand any movement of a part requires displacement of another part. Most of them have noterminations or points of attachment, and in hollow organs such as intestines, bloodvessels and uro-genital tract they usually form structures closed onto themselves, such asrings or bag-like containers In these situations, changes in the size of the lumen isachieved very efficiently by a concentric inward enlargement that accompanies musclecontraction. The longitudinal arrangement of collagen blocks an elongation of small bloodvessels upon contraction, further enhancing the efficiency of lumen reduction. In othermuscles, links between layers and special arrangements of the stroma allow both shorteningand elongation of a tubular organ to occur. The mechanics of smooth muscles has manycharacteristic features (some unique, some shared with those of hydrostats, some atvariance with other muscles) and histological data are a contribution to our understandingof these properties.

The c-Kit receptor tyrosine kinase regulates the development and differentiation ofseveral progenitor cells. In the gastrointestinal (GI) tract, the c-Kit regulates thedevelopment of the interstitial cells of Cajal (ICC) that are responsible for motilityregulation of the GI musculature. W-sash(Wsh) is an inversion mutation upstream of thec-kit promoter region that affects a key regulatory element, resultingin cell-type-specific altered gene expression, leading to a decrease in the number of mastcells, melanocytes, and ICC. We extensively examined the GI tract ofWsh/Wsh mice usingimmunohistochemistry and electron microscopy. Although the musculature of theWsh/Wsh mice did not show anyc-Kit immunoreactivity, we detected intensive immunoreactivity for transmembrane member16A (TMEM16A, anoctamin-1), another ICC marker. TMEM16A immunopositive cells were observedas ICC-MY in the gastric corpus-antrum and the large intestine, ICC-DMP in the smallintestine, and ICC-SM in the colon. Electron microscopic analysis revealed these cells asICC from their ultrastructural features, such as numerous mitochondria and caveolae, andtheir close contact with nerve terminals. In the developmental period, we examined 14.5and 18.5 day embryos but did not observe c-Kit immunoreactivity in theWsh/Wsh small intestine. Fromthis study, ICC subtypes developed and maturated structurally without c-Kit expression.Wsh/Wsh mice are a new modelto investigate the effects of c-Kit and unknown signaling on ICC development andfunction.

Background: Ex situ analyses of human myometrial tissue has been used to investigate the regulation of uterine quiescence and transition to a contractile phenotype. Following concerns about the validity of cultured primary cells, we examined whether myometrial tissue undergoes culture-induced changes ex situ that may affect the validity of in vitro models. Objectives: To determine whether human myometrial tissue undergoes culture-induced changes ex situ in Estrogen receptor 1 (ESR1), Prostaglandin-endoperoxide synthase 2 (PTGS2) and Oxytocin receptor (OXTR) expression. Additionally, to determine whether culture conditions approaching the in vivo environment influence the expression of these key genes. Methods: Term non-laboring human myometrial tissues were cultured in the presence of specific treatments, including; serum supplementation, progesterone and estrogen, cAMP, PMA, stretch or NF-κB inhibitors. ESR1, PTGS2 and OXTR mRNA abundance after 48 h culture was determined using quantitative RT-PCR. Results: Myometrial tissue in culture exhibited culture-induced up-regulation of ESR1 and PTGS2 and down-regulation of OXTR mRNA expression. Progesterone prevented culture-induced increase in ESR1 expression. Estrogen further up-regulated PTGS2 expression. Stretch had no direct effect, but blocked the effects of progesterone and estrogen on ESR1 and PTGS2 expression. cAMP had no effect whereas PMA further up-regulated PTGS2 expression and prevented decline of OXTR expression. Conclusion: Human myometrial tissue in culture undergoes culture-induced gene expression changes consistent with transition toward a laboring phenotype. Changes in ESR1, PTGS2 and OXTR expression could not be controlled simultaneously. Until optimal culture conditions are determined, results of in vitro experiments with myometrial tissues should be interpreted with caution.

Type 2 diabetic men commonly experience erectile dysfunction for whichphosphodiesterase-5 (PDE5) inhibitors like sildenafil (Viagra) are often recommended. Bypreventing degradation of cyclic guanosine monophosphate (cGMP) in vascular smooth muscle,these inhibitors also enhance arterial vasorelaxant effects of nitric oxide donors (whichstimulate cGMP synthesis). In the present work, we confirmed this enhancing effect afterco-administration of sildenafil with nitroprusside to freshly-isolated rat tail arterialtissues. However, in the same tissues we also observed that sildenafil does not enhancebut rather attenuates vasorelaxant effects of three commonly-used antidiabetic drugs, i.e.the biguanide metformin and the thiazolidinediones pioglitazone and rosiglitazone. Indeed,sildenafil completely blocked vasorelaxant effects of low concentrations of these drugs.In addition, we found that this same novel anti-vasorelaxant interaction of sildenafilwith these agents was abolished by either 1) omitting extracellular glucose or 2)inhibiting specific smooth muscle glycolytic pathways; pathways known to preferentiallyutilize extracellular glucose to fuel certain adenosine triphosphate (ATP)-dependent iontransporters: e.g. ATP-sensitive K channels, sarcoplasmic reticulum Ca-ATPase, plasmamembrane Ca-ATPase and Na/K-ATPase. Accordingly, we suspect that altered activity of oneor more of these ion transporters mediates the observed attenuating (anti-vasorelaxant)interaction of sildenafil with the antidiabetic drugs. The present results are relevantbecause hypertension is so common and difficult to control in Type 2 diabetes. The presentdata suggest that sildenafil might interfere with the known antihypertensive potential ofmetformin and the thiazolidinediones. However, they do not suggest that it will interactwith them to cause life-threatening episodes of severe hypotension, as can occur when itis co-administered with nitrates.