Contraction Of Intestinal Smooth Muscle Research Articles

Revealing the material basis and mechanism for the inhibition of intestinal peristalsis by Zingiber officinale Roscoe through integrated metabolomics, serum pharmacochemistry, and network pharmacology.

Abnormal relaxation and contraction of intestinal smooth muscle can cause various intestinal diseases. Diarrhea is a common and important public health problem worldwide in epidemiology. Zingiber officinale Roscoe (fresh ginger) has been found to treat diarrhea, but the material basis and mechanism of action that inhibits intestinal peristalsis remain unclear. Metabolomics and serum pharmacology were used to identify differential metabolites, metabolic pathways, and pharmacodynamic substances, and were then combined with network pharmacology to explore the potential targets of ginger that inhibit intestinal peristalsis during diarrhea treatment, and the targets identified were verified using molecular docking and molecular dynamic simulation. We found that 25 active components of ginger (the six most relevant components), 35 potential key targets (three core targets), 40 differential metabolites (four key metabolites), and four major metabolic pathways were involved in the process by which ginger inhibits intestinal peristalsis during diarrhea treatment. This study reveals the complex mechanism of action and pharmacodynamic material basis of ginger in the inhibition of intestinal peristalsis, and this information helps in the development of new Chinese medicine to treat diarrhea and lays the foundation for the clinical application of ginger.

Stroke Alters the Function of Enteric Neurons to Impair Smooth Muscle Relaxation and Dysregulates Gut Transit.

Gut dysmotility is common after ischemic stroke, but the mechanism underlying this response is unknown. Under homeostasis, gut motility is regulated by the neurons of the enteric nervous system that control contractile/relaxation activity of muscle cells in the gut wall. More recently, studies of gut inflammation revealed interactions of macrophages with enteric neurons are also involved in modulating gut motility. However, whether poststroke gut dysmotility is mediated by direct signaling to the enteric nervous system or indirectly via inflammatory macrophages is unknown. We examined these hypotheses by using a clinically relevant permanent intraluminal midcerebral artery occlusion experimental model of stroke. At 24 hours after stroke, we performed invivo and exvivo gut motility assays, flow cytometry, immunofluorescence, and transcriptomic analysis. Stroke-induced gut dysmotility was associated with recruitment of muscularis macrophages into the gastrointestinal tract and redistribution of muscularis macrophages away from myenteric ganglia. The permanent intraluminal midcerebral artery occlusion model caused changes in gene expression in muscularis macrophages consistent with an altered phenotype. While the size of myenteric ganglia after stroke was not altered, myenteric neurons from post-permanent intraluminal midcerebral artery occlusion mice showed a reduction in neuronal nitric oxide synthase expression, and this response was associated with enhanced intestinal smooth muscle contraction exvivo. Finally, chemical sympathectomy with 6-hydroxydopamine prevented the loss of myenteric neuronal nitric oxide synthase expression and stroke-induced slowed gut transit. Our findings demonstrate that activation of the sympathetic nervous system after stroke is associated with reduced neuronal nitric oxide synthase expression in myenteric neurons, resulting in impaired smooth muscle relaxation and dysregulation of gut transit.

Pico145 inhibits TRPC4-mediated mICAT and postprandial small intestinal motility

In intestinal smooth muscle cells, receptor-operated TRPC4 are responsible for the majority of muscarinic receptor cation current (mICAT), which initiates cholinergic excitation-contraction coupling. Our aim was to examine the effects of the TRPC4 inhibitor Pico145 on mICAT and Ca2+ signalling in mouse ileal myocytes, and on intestinal motility. Ileal myocytes freshly isolated from two month-old male BALB/c mice were used for patch-clamp recordings of whole-cell currents and for intracellular Ca2+ imaging using Fura-2. Functional assessment of Pico145’s effects was carried out by standard in vitro tensiometry, ex vivo video recordings and in vivo postprandial intestinal transit measurements using carmine red. Carbachol (50 µM)-induced mICAT was strongly inhibited by Pico145 starting from 1 pM. The IC50 value for the inhibitory effect of Pico145 on this current evoked by intracellularly applied GTPγS (200 µM), and thus lacking desensitisation, was found to be 3.1 pM, while carbachol-induced intracellular Ca2+ rises were inhibited with IC50 of 2.7 pM. In contrast, the current activated by direct TRPC4 agonist (-)-englerin A was less sensitive to the action of Pico145 that caused only ∼43 % current inhibition at 100 pM. The inhibitory effect developed rather slowly and it was potentiated by membrane depolarisation. In functional assays, Pico145 produced concentration-dependent suppression of both spontaneous and carbachol-evoked intestinal smooth muscle contractions and delayed postprandial intestinal transit. Thus, Pico145 is a potent GI-active small-molecule which completely inhibits mICAT at picomolar concentrations and which is as effective as trpc4 gene deficiency in in vivo intestinal motility tests.

Mechanisms of the White Sea cod Gadus morhua marisalbi (Gadidae) intestinal smooth muscle cholinergic contraction: the contribution of various subtypes of M-cholinergic receptors and Rho-kinase

The study of the functioning of various body systems in diff erent vertebrates is one of the key tasks of comparative physiology. Teleost fi sh are a large group of vertebrates, however, the mechanisms of functioning of their digestive tract have been studied little and mainly only for such a model object as Danio rerio. The aim of this work was to characterize the participation of various subtypes of M-cholinergic receptors and the enzyme Rho-kinase in the cholinergic contraction of the intestinal smooth muscles of the White Sea cod Gadus morhua marisalbi (Gadidae). A longitudinal strip was excised from the proximal cod intestine and placed in an apparatus for recording contractile responses in the isometric mode. Contractile responses to acetylcholine were ompletely blocked by atropine. Blockade of M3 cholinergic receptors with 4-DAMP resulted in a decrease in acetylcholine-induced contraction compared with the control response. Blockade of M1 receptors with pirenzepine led to a weakening of contraction, less pronounced than with blockade of M3 cholinergic receptors. Blockade of M2-cholinergic receptors with methoctramine did not affect the magnitude of the contractile response. Incubation of preparations with the Rho-kinase inhibitor fasudil was accompanied by a significant decrease in contractile responses compared with the control, as well as a faster decrease in the contraction force after reaching the “peak” of the reaction. Thus, acetylcholine causes contraction of cod intestinal smooth muscle by activating M3- and M1- but not M2-cholinergic receptors. The activity of the Rho-kinase enzyme contributes to the development and maintenance of cod intestinal smooth muscle contraction under the action of acetylcholine. The results obtained are of interest for comparative physiology, may be important for understanding the mechanisms of the damaging effect of environmental factors on the bony fish’ body, as well as for the use of fish as objects of preclinical studies of drugs.

Trans-ε-viniferin as an inhibitor of TMEM16A preventing intestinal smooth muscle contraction

TMEM16A regulator is an important tool to study the physiological functions and pathogenesis related to TMEM16A. In the present study, trans-ε-viniferin (TV) was identified as a TMEM16A inhibitor with inhibitory activity against TMEM16A mediated Cl− currents, which was reversible, without affecting intracytoplasmic Ca2+ concentration and TMEM16A protein expression. TV inhibited intestinal peristalsis and prolonged gastrointestinal transport time. TV could inhibit autonomic and Eact-stimulated intestinal contractility, and was equally effective in ACh- and HA-induced high contractile states. The results indicate that TV significantly inhibits the intestinal smooth muscle contraction, which may be applied in the treatment of TMEM16A-related intestinal dynamic abnormalities.

Integrating Network Pharmacology and In Vivo Model to Investigate the Mechanism of Biheimaer in the Treatment of Functional Dyspepsia.

Objective Biheimaer (BHM) is a hospital formulation for clinical treatment of dyspepsia and acid reflux, based on Compatibility Theory of Traditional Chinese Medicine. This study anticipated to elucidate the molecular mechanism of BHM against Functional dyspepsia via combined network pharmacology prediction with experimental verification. Methods Based on network pharmacology, the potential active components and targets of BHM in the treatment of functional dyspepsia were explored by prediction and molecular docking technology. The results of protein–protein interaction analysis, functional annotation, and pathway enrichment analysis further refined the main targets and pathways. The molecular mechanism of BHM improving functional dyspepsia mice induced by L-arginine + atropine was verified on the basis of network pharmacology. Results In this study, 183 effective compounds were screened from BHM; moreover, 1007 compound-related predicted targets and 156 functional dyspepsia-related targets were found. The results of enrichment analysis and in vivo experiments showed that BHM could regulate intestinal smooth muscle contraction to play a therapeutic role in functional dyspepsia by reducing the expression of NOS3, SERT, TRPV1, and inhibiting the inflammatory cytokine (IL-1β, TNF-α) to intervene the inflammatory response in mice. Conclusions This study revealed the molecular biological mechanisms of the Traditional Chinese Medicine formulation of BHM in functional dyspepsia by network pharmacology and experimental verification, meanwhile provided scientific support for subsequent clinical medication.

An Experimental Study on Intestine Transmission Function in SD Rats With Slow Transit Constipation After Electro-acupuncture Treatment in BL32 Acupoints

Objective: This study aims to use loperamide by gavage in SD rats to establish the model of STC, and evaluate the degree of similarity between the model and clinical diseases. Furthermore, the effect of electro-acupuncture when stimulating BL32 acupoints in SD rats with slow transit constipation were observed to determine changes in intestinal transmission function. Methods: Thirty SD rats were selected and divided randomly into three groups. Rats in the model group were given 2 mg/kg/d of loperamide tablets by gavage for 15 days. The morphology of the feces of rats was observed, and the dry weight of the feces and the time of first black feces movement with black ink by gavage after molding were measured. After 20 days of electro-acupuncture treatment at the BL32 acupoint, the time of first black feces movement, the ratio of black ink length of the entire length of the intestine, and contraction amplitude and frequency of contraction were measured. Results: The time of black feces movement in the two model groups was significantly prolonged (P <0.05). Furthermore, fecal grains, dry weight and moisture content were significantly reduced (P <0.05). On the 10th and 20th day of treatment, the time of black feces movement in the treatment group significantly decreased (P <0.05). The ratio of black ink length of the entire length of the intestine in the electro-acupuncture group was significantly higher than that in the model group (P <0.05). Moreover, contraction amplitude was significantly higher than in the model control group (P <0.05). Conclusion: Through loperamide gavage, the model of slow transit constipation in SD rats was successfully established in a relatively short period of time. Electro-acupuncture treatment had a rapid onset of action, and its mechanism might have been caused through strengthening the ability of intestinal smooth muscle contraction, and are independent with increased smooth muscle contraction frequency.

Cinobufagin Exerts Anticancer Activity in Oral Squamous Cell Carcinoma Cells through Downregulation of ANO1.

Anoctamin1 (ANO1), a calcium-activated chloride channel, is frequently overexpressed in several cancers, including oral squamous cell carcinoma (OSCC). OSCC is a highly aggressive cancer and the most common oral malignancy. ANO1 has been proposed as a potential candidate for targeted anticancer therapy. In this study, we performed a cell-based screening to identify novel regulators leading to the downregulation of ANO1, and discovered cinobufagin, which downregulated ANO1 expression in oral squamous cell carcinoma CAL-27 cells. ANO1 protein levels were significantly reduced by cinobufagin in a dose-dependent manner with an IC50 value of ~26 nM. Unlike previous ANO1 inhibitors, short-term (≤10 min) exposure to cinobufagin did not alter ANO1 chloride channel activity and ANO1-dependent intestinal smooth muscle contraction, whereas long-term (24 h) exposure to cinobufagin significantly reduced phosphorylation of STAT3 and mRNA expression of ANO1 in CAL-27 cells. Notably, cinobufagin inhibited cell proliferation of CAL-27 cells expressing high levels of ANO1 more potently than that of ANO1 knockout CAL-27 cells. In addition, cinobufagin significantly reduced cell migration and induced caspase-3 activation and PARP cleavage in CAL-27 cells. These results suggest that downregulation of ANO1 by cinobufagin is a potential mechanism for the anticancer effect of cinobufagin in OSCC.

Clinical Applications of Substance P (Neurokinin-1 Receptor) Antagonist in Canine Medicine.

Substance P binds to the Neurokinin-1 (NK-1) receptors found in the emetic center of the central nervous system (CNS) to induce emesis. Maropitant is a selective NK-1 receptor antagonist that inhibits the binding of substance P to NK-1 receptors and is commonly used to prevent and treat vomiting in dogs. This review study aimed to discuss and analyze the therapeutic potential of substance P (Neurokinin-1 receptor) antagonist with a particular focus on the drug maropitant in canine medicine. A systematic literature review was performed to identify the existing literature on the subject during the past 20 years (2001-2021) using such databases as ScienceDirect, PubMed, Scopus, and Google Scholar. The initial search identified 173 articles; however, 41 articles were selected for further analysis, based on the specific inclusion and exclusion criteria. Studies have already confirmed the role of substance P and NK-1 receptors in central pain processing, intestinal smooth muscle contraction, vasodilation, and neurogenic inflammation. Maropitant is one of the most effective veterinary antiemetic drugs that work well against peripheral and central stimuli that trigger the vomiting center. It has been already demonstrated that the therapeutic efficacy of maropitant for managing acute vomiting in dogs is associated with pancreatitis, gastritis, and parvoviral enteritis. It can also prevent and treat chemotherapy-induced emesis and delay the signs of nausea and adverse gastrointestinal effects. Regarding the broad-spectrum antiemetic activity of maropitant, it can be recommended for managing uremic vomiting in dogs. In addition, it has also exhibited an anesthetic sparing effect since the dogs treated with maropitant require a slightly lower percentage of isoflurane as an inhalational anesthetic. The NK-1 receptors are also identified in different areas of the pain pathways. Therefore, NK-1 receptor antagonists might be effective for managing visceral pain. However, further studies are required to establish the broad therapeutic potential of NK-1 receptor antagonist drugs, such as maropitant in canine medicine. It has been shown that the pain associated with the subcutaneous administration of maropitant is due to metacresol, a preservative used in some formulations. Therefore, the side effects can be eliminated by developing novel maropitant formulations specifically for dogs.

Enhanced Store-Operated Ca2+ Signal of Small Intestinal Smooth Muscle Cells Accelerates Small Bowel Transit Speed in Type 1 Diabetic Mouse.

Aims: The underlying mechanism of diabetic enteropathy, a common complication of type 1 diabetes, remains unclear. Store-operated Ca2+ entry (SOCE) is a ubiquitous type of Ca2+ influx involved in various cellular functions. Here, we show that SOCE-related stromal interaction molecule 1 (STIM1) and Orai1 participate in inappropriate cellular Ca2+ homeostasis, augmenting agonist-induced small intestinal smooth muscle contraction and small bowel transit speed in a mouse model of type 1 diabetes.Methods and Results: We used small interfering (si)RNA to suppress STIM1 and Orai1 proteins, and employed intracellular Ca2+, small intestinal contraction and intestinal transit speed measurement to investigate the functional change. We found that SOCE activity and Orai1 and STIM1 expression levels of small intestinal smooth muscle were significantly increased in cells cultured in high glucose medium or in diabetic mice. Gastrointestinal transit speed and SOCE-mediated contractions were markedly increased in diabetic mice; Knocking down Orai1 or STIM1 with siRNA rescued both alterations in diabetic mice. However, the Orai1-large conductance Ca2+-activated K+ (BKCa) channel interaction was decreased in diabetic mice, and suppressing Orai1 expression or inhibiting the BKCa channel increased agonist-induced small intestinal contractions in normal mice.Conclusion: We concluded that the increased SOCE caused by excessive STIM1 and Orai1 expression and decreased Orai1-BKCa interaction augmented small intestinal smooth muscle contraction and accelerated small bowel transit speed in diabetic mice. This finding demonstrates a pathological role for SOCE in diabetic enteropathy and provides a potential therapeutic target for diabetic enteropathy.

Nimodipine inhibits intestinal and aortic smooth muscle contraction by regulating Ca2+-activated Cl− channels

Nimodipine is a clinically used dihydropyridine L-type calcium channel antagonist that effectively inhibits transmembrane Ca2+ influx following the depolarization of smooth muscle cells, but the detailed effect on smooth muscle contraction is not fully understood. Ca2+-activated Cl− channels (CaCCs) in vascular smooth muscle cells (VSMCs) may regulate vascular contractility. We found that nimodipine can inhibit transmembrane protein 16A (TMEM16A) activity in a concentration-dependent manner by cell-based fluorescence-quenching assay and short-circuit current analysis, with an IC50 value of ~5 μM. Short-circuit current analysis also showed that nimodipine prevented Ca2+-activated Cl− current in both HT-29 cells and mouse colonic epithelia accompanied by significantly decreased cytoplasmic Ca2+ concentrations. In the absence of extracellular Ca2+, nimodipine still exhibited an inhibitory effect on TMEM16A/CaCCs. Additionally, the application of nimodipine to CFTR-expressing FRT cells and mouse colonic mucosa resulted in mild activation of CFTR-mediated Cl− currents. Nimodipine inhibited basolateral CCh-activated K+ channel activity with no effect on Na+/K+-ATPase activity. Evaluation of intestinal smooth muscle contraction showed that nimodipine inhibits intestinal smooth muscle contractility and frequency, with an activity pattern that was similar to that of non-specific inhibitors of CaCCs. In aortic smooth muscle, the expression of TMEM16A in thoracic aorta is higher than that in abdominal aorta, corresponding to stronger maximum contractility in thoracic aorta smooth muscle stimulated by phenylephrine (PE) and Eact. Nimodipine completely inhibited the contraction of aortic smooth muscle stimulated by Eact, and partially inhibited the contraction stimulated by PE. In summary, the results indicate that nimodipine effectively inhibits TMEM16A/CaCCs by reduction transmembrane Ca2+ influx and directly interacting with TMEM16A, explaining the mechanisms of nimodipine relaxation of intestinal and aortic smooth muscle contraction and providing new targets for pharmacological applications.

Anti-diarrheal therapeutic potential of diminazene aceturate stimulation of the ACE II/Ang-(1–7)/Mas receptor axis in mice: A trial study

The angiotensin (Ang) II converting enzyme (ACE II) pathway has recently been shown to be associated with several beneficial effects on the body, especially on the cardiac system and gastrointestinal tract. ACE II is responsible for converting Ang II into the active peptide Ang-(1–7), which in turn binds to a metabotropic receptor, the Mas receptor (MasR). Recent studies have demonstrated that Diminazene Aceturate (DIZE), a trypanosomicide used in animals, activates the ACE II pathway. In this study, we aimed to evaluate the antidiarrheal effects promoted by the administration of DIZE to activate the ACE II/Ang-(1–7)/MasR axis in induced diarrhea mice models. The results show that activation of the ACE II pathway exerts antidiarrheal effects that reduce total diarrheal stools and enteropooling. In addition, it increases Na+/K+-ATPase activity and reduces gastrointestinal transit and thus inhibits contractions of intestinal smooth muscle; decreases transepithelial electrical resistance, epithelial permeability, PGE2-induced diarrhea, and proinflammatory cytokines; and increases anti-inflammatory cytokines. Enzyme-linked immunosorbent assay (ELISA) demonstrated that DIZE, when activating the ACE II/Ang-(1–7)/MasR axis, can still interact with GM1 receptors, which reduces cholera toxin-induced diarrhea. Therefore, activation of the ACE II/Ang-(1–7)/MasR axis can be an important pharmacological target for the treatment of diarrheal diseases.

Effect of the leaves aqueous extract of Jodina rhombifolia (Hook. & Arn.) Reissek (Santalaceae) on intestinal function and its acute toxicity

Ethnopharmacological relevanceIn Argentine traditional medicine it is recorded the use of Jodina rhombifolia (Hook. & Arn.) Reissek (Santalaceae) leaves for treating various affections that compromise the intestinal normal functioning. Aim of the studyThe aim of this study was to assess the effect of J. rhombifolia leaves lyophilized aqueous extract on the intestinal function by means of in vivo and ex vivo experimental assays for determinate the antidiarrheal and antispasmodic capacity. Furthermore, was to evaluate its acute toxicity potential by oral and intraperitoneal administration of extract. Materials and methodsThe in vivo assays were conducted by the experimental techniques of intestinal transit in mice, intestinal fluid accumulation in rats and Castor oil-induced diarrhea in mice. In the ex vivo assays, isolated rat duodenum and ileum segments were used for to evaluate the antispasmodic activity through contractile concentration-response curves induced by Carbachol and CaCl2. The acute toxicity of the extract was also investigated by oral and intraperitoneal administration. ResultsThe extract intraperitoneal administration at the doses 125, 250 and 500 mg/kg, caused a marked reduction in the normal intestinal transit and in the number of diarrheal episodes in a dose-dependent manner. However, the extract did not produce significant changes in the intestinal fluid accumulation with any of the tested dose. The extract demonstrated a non-competitive inhibitory effect on the contractions of intestinal smooth muscle experimentally provoked by the spasmogenic agents Carbachol and CaCl2 in a dose-dependent manner (IC50 = 10.57 ± 1.38 and 6.29 ± 1.54 mg extract/ml Tyrode solution in the isolated organ bath, respectively). ANOVA indicated a significative effect of treatment (p < 0.001). ConclusionsThe scientific evidence achieved based in the used experimental models allows us to indicate that J. rhombifolia leaves lyophilized aqueous extract manifest an antispasmodic activity on intestinal smooth muscle without observation of apparent toxicity symptoms.

SUPPRESSION OF THE SMALL INTESTINE SHRINKABLE ACTIVITY AFTER USAGE OF THE ANESTHETIC KETAMINE

The work is devoted to the problem of general anesthetics side effects during surgical interventions. In particular, some of the common complications of anesthetics usage are intestinal motility disorders (eg, ileus), nausea and vomiting. We analyzed a clinical data of 60 patients who underwent analgosedation (AS) during coronary artery stenting. They were divided into 2 groups (1 group - AS with Diazepam and Fentanyl, 2 group - AS with Ketamine, Fentanyl and Propofol). Significantly more frequent manifestations of nausea in the perioperative period were identified in the group where ketamine was used. This formed the basis of the study of the molecular and cellular mechanisms of the effects of ketamine on the contractile activity of the smooth muscles of the small intestine.Anesthetics are known to interact with receptors, G-proteins and ion channels, including transient receptor potential channels (TRP channels). The member of this superfamily, TRPC4 channel, which is coupled to muscarinic receptors (M2/M3 type) through G-protein activation and causes cholinergic excitation and contraction of small intestinal smooth muscles, may be a potential target for ketamine.Thus, we aimed to investigate the effects of ketamine (100 μM) on the muscarinic cation current (mICAT), which underlies cholinergic excitation-contraction coupling of visceral smooth muscles. All experiments were performed on single ileal myocytes freshly isolated from the longitudinal layer of the mouse ileum using patch-clamp techniques in the whole-cell configuration. mICAT was recorded using symmetrical Cs+ solutions (containing Cs+125 mM). [Ca2+]i was ‘clamped’ at 100 nM using 10 mM BAPTA/4.6 mM CaCl2 mixture. Measurements of isometric contractile force of the small intestinal smooth muscles were recorded using tensiometry techniques. It was showed that 100 μM ketamine inhibits mICAT . mICAT initiated by the application of CCh (50 μM) was suppressed on 64% (n=5) and mICAT, induced by intracellular GTP γ s (200 μM) that interacts directly with the G-proteins (when muscarinic receptors are bypassed) was inhibited on 42% (n=5). Ketamine inhibited intestinal smooth muscle contractions evoked by carbachol (50 μM) by about 40 % (n=5). Thus, we can conclude that both muscarinic receptors and G-proteins (or their coupling) are affected by ketamine, but the main sites of ketamine action appear to be the G-proteins. These data will provide basis for the molecular mechanisms of postoperative motility disorders and thus may be important for the development of novel approaches to the correction of such states.

Se Deficiency Induced Inflammation Resulting to a Diminished Contraction of the Small Intestinal Smooth Muscle in Mice.

Selenium (Se) is an essential trace element for both humans and animals. Se deficiency leads to myocardial injury, reproductive disorder, increased exudation, inflammatory injury, and other diseases. The present study analyzed the relationships of Se deficiency, inflammation, and smooth muscle contraction in the small intestine, which is the main tissue that digests and absorbs Se. The model was established by feeding the animals diets with different concentrations of Se. The results showed that the dietary Se content was positively correlated with the blood Se concentration and the intestinal Se concentration. ROS and MPO activity increased with the lack of Se. TNF-α, IL-1β, and IL-6 expression was increased at both the mRNA and protein levels with Se deficiency. The pathways tested showed that the IκBα, NF-κB p65, p38, ERK, and JNK phosphorylation levels were significantly increased with the lack of Se. Moreover, the contractility analysis confirmed that contraction of the intestinal smooth muscle was attenuated by Se deficiency, as shown by the MedLab data acquisition system. These results further illuminated the relationship between inflammation and inhibition of smooth muscle contraction under Se deficiency in the small intestine. The Ca2+ concentration was decreased, and RhoA phosphorylation and ROCK expression were also inhibited by Se deficiency. The results also showed that MLC protein phosphorylation decreased with Se deficiency. In conclusion, the present study indicated that inflammation under Se deficiency leads to the inhibition of smooth muscle contraction in the small intestine.

Biopolymer Extracted from Anadenanthera colubrina (Red Angico Gum) Exerts Therapeutic Potential in Mice: Antidiarrheal Activity and Safety Assessment.

Anadenanthera colubrina var. cebil (Griseb.) Altschul (Fabaceae family), commonly known as the red angico tree, is a medicinal plant found throughout Brazil’s semi-arid area. In this study, a chemical analysis was performed to investigate the antidiarrheal activity and safety profile of red angico gum (RAG), a biopolymer extracted from the trunk exudate of A. colubrina. Upon FT-IR spectroscopy, RAG showed bands in the regions of 1608 cm−1, 1368 cm−1, and 1029 cm−1, which relate to the vibration of O–H water molecules, deformation vibration of C-O bands, and vibration of the polysaccharide C-O band, respectively, all of which are relevant to glycosidic bonds. The peak molar mass of RAG was 1.89 × 105 g/mol, with the zeta potential indicating electronegativity. RAG demonstrated high yield and solubility with a low degree of impurity. Pre-treatment with RAG reduced the total diarrheal stool and enteropooling. RAG also enhanced Na+/K+-ATPase activity and reduced gastrointestinal transit, and thereby inhibited intestinal smooth muscle contractions. Enzyme-Linked Immunosorbent Assay (ELISA) demonstrated that RAG can interact with GM1 receptors and can also reduce E. coli-induced diarrhea in vivo. Moreover, RAG did not induce any signs of toxicity in mice. These results suggest that RAG is a possible candidate for the treatment of diarrheal diseases.

Pediatric Intestinal Pseudo-obstruction in the Era of Genetic Sequencing.

The purpose of this review is to discuss current knowledge on pediatric intestinal pseudo-obstruction. We will also review new mutations that have been identified through advancement in genetic testing, allowing for a better understanding of the underlying mechanisms of intestinal dysmotility and potential etiologies. With the advancements in genetic testing, new mutations have been identified in the diagnosis of megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS), a disorder leading to pediatric pseudo-obstruction. MYLK, LMOD1, MYL9, and MYH11 encode for various proteins within smooth muscle cells; abnormalities within these proteins lead to abnormal intestinal smooth muscle contractions. Chronic intestinal pseudo-obstruction (CIPO) is defined by symptoms of bowel obstruction in the absence of a lumen-occluding lesion. CIPO is a heterogeneous group of disorders caused by abnormalities in the enteric neurons, intestinal smooth muscle, and/or the interstitial cells of Cajal (ICC). Symptoms can be non-specific and etiologies include both primary and secondary causes of CIPO that contribute to the delay in recognizing this condition and making the correct diagnosis. Chronic intestinal pseudo-obstruction has been recognized in both adults and children with fundamental differences in the etiology, symptom onset, clinical features and natural history of this disorder. For this reason, it has been considered a separate entity referred to as pediatric intestinal pseudo-obstruction (PIPO).

Interstitial cell network volume is reduced in the terminal bowel of ageing mice.

Ageing is associated with impaired neuromuscular function of the terminal gastrointestinal (GI) tract, which can result in chronic constipation, faecal impaction and incontinence. Interstitial cells of cajal (ICC) play an important role in regulation of intestinal smooth muscle contraction. However, changes in ICC volume with age in the terminal GI tract (the anal canal including the anal sphincter region and rectum) have not been studied. Here, the distribution, morphology and network volume of ICC in the terminal GI tract of 3‐ to 4‐month‐old and 26‐ to 28‐month‐old C57BL/6 mice were investigated. ICC were identified by immunofluorescence labelling of wholemount preparations with an antibody against c‐Kit. ICC network volume was measured by software‐based 3D volume rendering of confocal Z stacks. A significant reduction in ICC network volume per unit volume of muscle was measured in aged animals. No age‐associated change in ICC morphology was detected. The thickness of the circular muscle layer of the anal sphincter region and rectum increased with age, while that in the distal colon decreased. These results suggest that ageing is associated with a reduction in the network volume of ICC in the terminal GI tract, which may influence the normal function of these regions.

Study on effects of electrical stimulation on rabbit esophageal body motility in vivo.

Electric stimulation (ES) could induce contraction of intestinal smooth muscle. The aim of this study was to analyze the effects of ES on esophageal motility and the underlying mechanism in vivo. Twenty-eight rabbits were equipped with a pair of subserosa electrodes (connected to an electrical stimulator) in the lower segment of the esophagus. The ES signal consisted of bipolar rectangular pulse trains, lasting for 3 s, with different amplitudes (1 mA, 3 mA, 5 mA and 10 mA), and frequencies (10 Hz, 20 Hz and 50 Hz). The amplitude of the contraction was recognized by high-resolution manometry. The effect of ES was tested under anesthesia and following administration of atropine, phentolamine or L-NAME. ES induced esophageal contraction at the stimulated site. A statistically significant increase in esophageal pressure was observed when the stimulation amplitude was above 3 mA. The increase in esophageal pressure was associated with the amplitude of stimulus as well as the frequency. During stimulation, atropine, phentolamine and L-NAME had no effect on the increase of esophageal pressure induced by ES. These findings implied that ES induced esophageal contraction were not mediated via the NANC, adrenergic or cholinergic pathway. The amplitude of esophageal contraction was current and frequency dependent.

The IgCAM CLMP regulates expression of Connexin43 and Connexin45 in intestinal and ureteral smooth muscle contraction in mice.

ABSTRACTCAR-like membrane protein (CLMP), an immunoglobulin cell adhesion molecule (IgCAM), has been implicated in congenital short-bowel syndrome in humans, a condition with high mortality for which there is currently no cure. We therefore studied the function of CLMP in a Clmp-deficient mouse model. Although we found that the levels of mRNAs encoding Connexin43 or Connexin45 were not or were only marginally affected, respectively, by Clmp deficiency, the absence of CLMP caused a severe reduction of both proteins in smooth muscle cells of the intestine and of Connexin43 in the ureter. Analysis of calcium signaling revealed a disordered cell-cell communication between smooth muscle cells, which in turn induced an impaired and uncoordinated motility of the intestine and the ureter. Consequently, insufficient transport of chyme and urine caused a fatal delay to thrive, a high rate of mortality, and provoked a severe hydronephrosis in CLMP knockouts. Neurotransmission and the capability of smooth muscle cells to contract in ring preparations of the intestine were not altered. Physical obstructions were not detectable and an overall normal histology in the intestine as well as in the ureter was observed, except for a slight hypertrophy of smooth muscle layers. Deletion of Clmp did not lead to a reduced length of the intestine as shown for the human CLMP gene but resulted in gut malrotations. In sum, the absence of CLMP caused functional obstructions in the intestinal tract and ureter by impaired peristaltic contractions most likely due to a lack of gap-junctional communication between smooth muscle cells.