Castor Oil-induced Diarrhea In Mice Research Articles

Pickle water ameliorates castor oil-induced diarrhea in mice by regulating the homeostasis of the gut microbiota and intestinal mucosal barrier

Pickle water ameliorates castor oil-induced diarrhea in mice by regulating the homeostasis of the gut microbiota and intestinal mucosal barrier

Antispasmodic and antidiarrheal effects of Juniperus oxycedrus L. on the jejunum in rodents.

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.

Antidiarrheal Activities of the Methanol Leaf Extracts of Olinia rochetiana (Oliniaceae) Against Castor Oil-Induced Diarrhea in Mice.

Olinia rochetiana has been used traditionally to cure diarrheal disease. Therefore, this study aimed to investigate the acute toxicity and antidiarrheal effect of O. rochetiana leaf extracts. Cold maceration was used to extract plant leaf powder with 80% methanol. The extract's antidiarrheal action was tested against a castor oil-induced diarrheal model, a charcoal meal test, and enteropooling tests at doses of 100, 200, and 400 mg/kg. Negative controls received the vehicle at 10 mL/kg, while positive controls received loperamide at 3 mg/kg. From the study, no apparent toxicity was observed when a single dose of 2000 mg/kg was administered. In the castor oil-induced model, the extract delayed the onset of diarrhea, reduced stool frequency, and decreased wet feces weight and number in a dose-dependent manner at 200 mg/kg (p < 0.05) and 400 mg/kg (p < 0.01). The percent reduction in moist feces at 100, 200, and 400 mg/kg was 54.2, 23.97, and 18.26%, respectively, indicating a significant dose-dependent decrease. In a charcoal meal test, the extracts at 200 and 400 mg/kg revealed a peristaltic index of 65 and 46%, respectively, with considerable inhibition of charcoal transport at 23 and 39%. The weight and volume of intestinal contents dropped significantly at a dose of 400 mg/kg (p < 0.01), which is 0.43 mg/kg, in the enteropooling test when compared with the tested dose. The computed in vivo antidiarrheal index revealed diarrheal inhibition values of 46.06 and 71.06% at 200 and 400 mg/kg, respectively. In the current investigation, O. rochetiana showed significant antidiarrheal activity with no symptoms of toxicity in mice.

Phytochemical and in Vivo Biological Studies of Tanaecium Bilabiatum (Sprague) L.G. Lohmann

Phytochemical analysis of Tanaecium bilabiatum leaf led to the isolation and identification of salacinin C, daturadiol and a mixture of triterpenes, α-amyrin and β-amyrin. Besides chemical analysis, the methanol extract of T. bilabiatum leaf, METBL was evaluated for analgesic, anti-diarrheal and hypoglycemic potential in mice model. For analgesic activity test by tail immersion technique, the extract significantly extended the percent elongation time of thermal nociception. METBL (400 mg/kg bw) also established the highest inhibition of formalin-induced abdominal writhing (69.39%) in mice, which was comparable with the standard aspirin (75.51%). Likewise, dose-dependent antidiarrheal activity was observed in this test. After oral administration, METBL demonstrated maximum inhibition (40.93%) of castor oil-induced diarrhea in mice. In oral glucose tolerance test, the plant extract produced significant (p &lt; 0.05) hypoglycemic activity. Therefore, it is concluded that T. bilabiatum is a promising source of bioactive compounds with analgesic, antidiarrheal and hypoglycemic potential. Bangladesh J. Bot. 52(1): 119-127, 2023 (March)

Antidiarrheal activity of the extracts of Valeriana jatamansi Jones on castor oil-induced diarrhea mouse by regulating multiple signal pathways

Ethnopharmacological relevanceValeriana jatamansi Jones, a traditional medicine, is used for various medicinal purposes worldwide. This species is popular for its gastro-protective properties and has been verified to exert antidiarrheal effects. Qiuxieling mixture, an oral liquid preparation used to treat diarrhea in children in clinical practice, was extracted from V. jatamansi Jones.Aim of the study: Although Qiuxieling mixture has a good preventive effect on diarrhea children, the disgusting smell makes it intolerable. Therefore, we extracted odorless products from V. jatamansi Jones and Qiuxieling mixture. The present study is aimed to investigate the protective effects of two ethanolic extracts of V. jatamansi Jones and Qiuxieling mixture against castor oil-induced diarrhea and their possible mechanisms in mice. Materials and methodsThe two extracts of V. jatamansi Jones and Qiuxieling mixture were detected by HPLC. A castor oil-induced diarrheal model was used to evaluate the antidiarrheal effects. The expression of Occludin in the small intestine was measured by IHC. Western blotting and immunofluorescence were used to detect the expression of proteins related to the oxidative stress and GSDMD-mediated pyroptosis signaling pathways. ELISA was used to detect the expression of IL-6 and IL-1β in the small intestine of mice with diarrhea. ResultsThe two extracts of V. jatamansi Jones and Qiuxieling mixture dose-dependently reduced the diarrhea index and the diarrhea rate, delayed the onset of diarrhea, and decreased the weight of the intestinal content. Meanwhile, they reversed the decreased expression of Occludin and restored the activity of Na+-K+-ATPase in the intestines of diarrheal mice. In addition, they reversed the depletion of GSH, attenuated the activation of the ERK/JNK pathway, promoted the Nrf2/SOD1 signaling pathways, and decreased the release of ROS in the intestines of diarrheal mice. Moreover, they suppressed GSDMD-mediated pyroptosis by downregulating the NLRP3/caspase-1/GSDMD signaling pathway. ConclusionsThe two extracts of V. jatamansi Jones and Qiuxieling mixture exerted protective effects on castor oil-induced diarrhea in mice through a variety of mechanisms, including antioxidant stress, restoration of tight junctions between intestinal mucosal cells and regulation of the GSDMD-mediated pyroptosis pathway.

EVALUATION OF THE IN VIVO AND IN VITRO ANTIDIARRHEAL EFFECTS OF METHANOL LEAF EXTRACTS OF Boscia salicifolia IN MICE AND ISOLATED RABBIT JEJUNUM

Diarrhoea is one of the leading causes of preventable death in developing countries andmainly affects children and infants. The leaves of Boscia salicifolia has been used as anherbal remedy in the treatment of diarrhea in many parts of Northern Nigeria and WestAfrica. Thus, this study was aimed at investigating this enthnomedical claims of the plantextract on the gastrointestinal tracts. The fresh leaves were collected from Kalshingi townof Akko Local Government area Gombe State in August. They were dried for a weekunder the sun and the grounded into powder. Six hundred (600 g) grams of the powderedleaves were subjected to successive cold extraction with 80 % methanol. The extract wasused for acute toxicity, phytochemical screening, antidiarrheal and spasmolyticevaluation of acetylcholine, histamine and serotonin induced contractions on rabbitjejunum. The preliminary phytochemical screening revealed the presence of alkaloids,flavonoids, tannins, saponins, terpenes, steroid and glycosides. The effect methanol leafextract of B. salicifolia on castor oil-induced diarrhoea in mice showed a significantreduction (p&lt;0.05) in the diarrheal drops with increase percentage inhibition of diarrhea45.95, 40.54, and 29.72% at 380, 760, and 1140 mg/kg doses of the extract, respectively.The standard drug (loperamide 5 mg/kg) inhibited diarrhoeal droppings by 51 %. Theextract significantly (p&lt;0.001) decreases the force of rabbit ileum contraction induced byacetylcholine, histamine and serotonin at 7 x 10-10 g/ml, 1 x 10-7 g/ml and 1 x 10-7 g/mlrespectively. The methanol leaf extract of Boscia salicifolia was found to be relativelysafe and possess significant anti-diarrhoeal activity, and this justifies the use of this plantin the treatment of diarrhoea in the traditional settings.Keywords: Boscia salicifolia, diarrhea, in vivo, in vitro, spasmolytic, castor oil

Limonitum Ameliorates Castor Oil-Induced Diarrhoea in Mice by Modulating Gut Microbiota

Diarrhoea is a common clinical condition; its pathogenesis is strongly associated with gut microbiota dysbiosis. Limonitum is a well-known traditional Chinese medicine that exerts appreciable benefits regarding the amelioration of diarrhoea. However, the mechanism through which Limonitum ameliorates diarrhoea remains unclear. Here, the efficacy and underlying mechanism of Limonitum decoction (LD) regarding diarrhoea were explored from the aspect of gut microbiota. Castor oil (CO) was used to induce diarrhoea in mice, which were then used to evaluate the effects of LD regarding the timing of the first defecation, diarrhoea stool rate, degree of diarrhoea, diarrhoea score, intestinal propulsive rate, and weight of intestinal contents. The concentrations of short-chain fatty acids (SCFAs), including acetic, propionic, isobutyric, butyric and valeric acids, were analysed by gas chromatography-mass spectrometry (GC-MS). The 16S rRNA high-throughput sequencing technology was applied to evaluate changes in the gut microbiota under exposure to LD. LD was found to effectively ameliorate the symptoms of diarrhoea, and the diversity and relative abundance of gut microbiota were restored to normal levels following LD treatment. Additionally, LD significantly restored the observed reductions in SCFAs. These results provide strong evidence that LD can sufficiently ameliorate diarrhoea in mice by regulating their gut microbiota. The findings presented here highlight that Limonitum may constitute a prospective remedy for diarrhoea.

In vivo anti-diarrheal activity of jujube honey on castor oil-induced diarrhea in mice.

To evaluate the antidiarrheal effect of jujube honey on experimentally castor oil-induced diarrhea in mice by using different testing models (diarrhea, enteropooling and gastrointestinal motility). The mice intragastric administration castor oil was post-treated after 30 min with jujube honey, diluted jujube honey and loperamide or vehicles in different experimental groups. The onset and number of wet defecation on the absorbent paper was recorded for each animal for 4 h. Plasma was examined for C-reactive protein (CRP) and nitric oxide (NO) for clinical inflammation evaluation. The oxidative stress was investigated by superoxide dismutase (SOD) and catalase levels. The diluted jujube honey exhibited an important antidiarrheal activity manifested by significant delay in onset of diarrhea (P < 0.05), loss in number of wet stools (P < 0.001), total number of stools (P < 0.001) and total stool weight of fecal output (P < 0.001) in 4 h in castor oil-induced diarrheal groups. The inhibition of intestinal transit of charcoal meal (P < 0.01) is a most likely mechanism that may account for antidiarrheal effect of jujube honey. It also significantly increased SOD and catalase levels (P < 0.001) and significantly decreased CRP and NO in plasma diarrheic mice (P < 0.01). The results of this research reveal that the jujube honey contains pharmacologically active compounds with antidiarrheal properties. However, diluted honey is more effective in the treatment of diarrhea and imbalance of intestinal motility. In conclusion, these findings illustrated the antidiarrheal activity effect of jujube honey and which has the strongest evidence supporting its use in the treatment of diarrhea in traditional medicine.

Preparations from Campomanesia reitziana reduce the gastrointestinal motility and castor oil-induced diarrhea in a non-opioid and non-dopaminergic pathway in mice and display antimicrobial activity in vitro.

This study investigated the antidiarrheal potential of the aqueous extract (AECR) and hydroalcoholic extract ofCampomanesia reitzianaleaves (HECR), its ethyl acetate (EAF) and dichloromethane fractions (DCMF), and myricitrin isolated from EAF. The total phenols and flavonoids were measured, followed by chromatography and myricitrin isolation. The 2,2-diphenyl-1-picryl-hydrazyl scavenger activity, the cytotoxicity, and the effects on LPS-induced nitrite production in intestinal epithelial cells (IEC-6) were quantified. The effect of HECR, EAF, DCMF, and AECR on intestinal motility (IT), gastric emptying (GE), and castor oil-induced diarrhea in mice was determined, as well as its antimicrobial activity. The administration of AECR 10% (10ml/kg, p.o), but not HECR (300mg/kg), reduced the GE and IT by 52 and 51%. The EAF and DCMF at 300mg/kg also reduced IT but did not change GE. Moreover, AECR and EAF, but not DCMF, inhibited the castor oil-induced diarrhea and naloxone or metoclopramide pretreatment did not change these effects. Myricitrin did not change IT and the evacuation index of mice. Finally, the dry residue of AECR inhibited bacterial growth and EAF showed bacteriostatic activity againstS.aureus,E.coli,andS.typhimuriumand antifungal forC.albicans. However, none of the preparations alter the viability ofGiardia spp. trophozoites. The AECR and EAF can be effective to treat diarrhea acting through opioid- or dopaminergic type 2 receptor-independent mechanisms and by its antimicrobial actions.

Evaluation of Analgesic, Antidiarrheal and Hypoglycemic Activities of &lt;i&gt;Wendlandia paniculata&lt;/i&gt; (Roxb.) DC Leaves Extract using Mice Model

The leaves of Wendlandia paniculata (Roxb.) DC is a rare folk medicine among tribal people even though there is no scientific literature available regarding its pharmacological potentials. This study aimed to evaluate the effects of crude methanol extract of W. paniculata (Roxb.) DC. Leaves in mice model. The in vivo glucose-lowering capacity of methanolic crude extract was analyzed utilizing the well-known tail tipping method. The central and peripheral analgesic activity was evaluated using the ‘tail flick' and writhing assay respectively. The anti-diarrheal activity was evaluated using the method of castor oil-induced diarrhea in mice. As a result, oral administration (400 mg/ kg) of the extract resulted significant (P&lt;0.001) delay in response to pain and inhibition of acetic acid-induced writhing response. The results were comparable to the respective standards morphine (2 mg/kg) and diclofenac (50 mg/kg). Likewise, maximum reduction (p&lt;0.05) of blood glucose (32.3%) was observed 180 min after oral intake(400 mg/kg) of the extract compared to the 47% of the standard glibenclamide. Moreover, a reduction of diarrheal feces was observed 45.83% (p&lt;0.001) compared to standard loperamide 70.83%. The in vivo bioassays confirmed that the crude methanol extract of leaves of W. paniculata possesses significant analgesic (central, peripheral) and antidiarrheal activity with mild glucose-lowering activity. Isolation of bioactive compounds is needed to justify the molecule responsible for the activity.

Tannins extract from Galla Chinensis can protect mice from infection by Enterotoxigenic Escherichia coli O101

BackgroundEnterotoxigenic Escherichia coli (ETEC) is classically associated with acute secretory diarrhea, which induces 2 million people death in developing countries over a year, predominantly children in the first years of life. Previously, tannins (47.75%) were extracted from Galla Chinensis and prepared as Galla Chinensis oral solution (GOS) which showed significant antidiarrheal activity in a castor oil-induced diarrhea in mice. Whether the tannins extract were also effective in treatment of ETEC-induced diarrhea was determined in this study.MethodsMice were randomly divided into 6 groups (n = 22). The mice in the normal and untreated groups were given normal saline. Three GOS-treated groups were received different concentrations of GOS (5, 10 and 15%, respectively) at a dose of 10 mL/kg. Mice in the positive control group were fed with loperamide (10 mg/kg). The treatment with GOS started 3 days before infection with ETEC and continued for 4 consecutive days after infection. On day 3, mice were all infected with one dose of LD50 of ETEC, except those in the normal group. Survival of mice was observed daily and recorded throughout the study. On days 4 and 7, samples were collected from 6 mice in each group.ResultsGOS could increase the survival rate up to 75%, while in the untreated group it is 43.75%. The body weights of mice treated with 15% GOS were significantly increased on day 7 in comparison with the untreated group and the normal group. GOS-treatment recovered the small intestine coefficient enhanced by ETEC-infection. The diarrhea index of mice treated with GOS was significantly decreased. GOS increased the levels of IgG and sIgA in the terminal ileum and decreased the levels of pro-inflammatory cytokines (IFN-γ, TNF-α, IL-1β, IL-6 and IL-8) in serum. GOS could increase the amount of intestinal probiotics, Lactobacilli and Bifidobacteria. GOS could alleviate colon lesions induced by ETEC-infection. GOS showed higher potency than loperamide.ConclusionsGOS could be a promising drug candidate for treating ETEC infections.

Antidiarrhoeal activity of the 80% methanol root extract of &lt;i&gt; Thalictrum ryhnchocarpum&lt;/i&gt; Dill. &amp; A. Rich and its major constituent against castor oil-induced diarrhoea in mice

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Phytochemical and Pharmacological Investigations of Lannea grandis (Dennst.) Engl. Leaves Extracts

Medicinal plants can serve as a prominent source of bioactive secondary metabolites which can exert beneficial effects to combat against many human diseases. Lannea grandis (Dennst.) Engl, a member of Anacardiaceae family has several applications in the ethnomedical practices. In this study, the ethanol extract of L. grandis leaf (ELG) along with its petroleum-ether and chloroform fractions were subjected to phytochemical analysis along with the evaluation of antimicrobial, anti-diarrheal, and sedative activities using standard protocols. Phytochemical screening was performed according to common phytochemical tests. Antimicrobial assay was carried out by disc diffusion method where the ethanol extract of L. grandis showed significant activity against tested bacterial species (zone of inhibitions = 10.8±0.85 to 13.8±1.84 mm). The same extract also exhibited the highest antifungal activity against Blastomyces dermatitidis (zone of inhibition = 20.5±0.35 mm). The chloroform fraction of L. grandis at the dose of 400 mg/kg body weight produced a significant (p&lt;0.05) anti-diarrheal effect with 40.69% inhibition of castor oil-induced diarrhea in mice. During sedative activity assay in mice, the plant extract at the dose of 400 mg/kg body weight produced mild reduction in the time of onset of sleep and increased the length of the sleeping time induced by a sedative, phenobarbitone. In conclusion, the plant L. grandis can be considered as a base for the development of new drugs and phytomedicine.&#x0D; Keywords: Lannea grandis, phytochemical, antimicrobial, anti-diarrheal, sedative activity.

Effects of hydroethanolic extract of Cameroonian propolis (Promax-c) on castor oil-induced diarrhea in mice

The aim of our work was to evaluate the effect of hydroethanolic extract of Cameroonian propolis (Promax-c) on castor oil-induced diarrhea in mice. Diarrhea was induced in mice by oral administration 0.5 mL of castor oil in all mice. To determine the effective doses, each mouse received, 30 minutes after the administration of castor oil, one of the single oral doses of Promax-c: 0, 37.5, 75, 150, and 300 mg/kg bw. The mass and frequency of stool were measured and recorded per hour for five hours. The effect of Promax-c on the intestinal motility was evaluated by measuring the distance traveled by the charcoal meal in thirty minutes. The effects of Promax-c on intestinal secretion were evaluated by measuring the volume of the intestinal content and by dosing the electrolytes (Na+, K+, and Cl-) in the intestinal content by the colorimetric method. Promax-c produced significant (P

Effects of hydroethanolic extract of Cameroonian propolis (Promax-c) on castor oil-induced diarrhea in mice

Effects of hydroethanolic extract of Cameroonian propolis (Promax-c) on castor oil-induced diarrhea in mice

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.

Analgesic, Antidiarrheal and CNS-depressant Activities of Flemingia macrophylla (Willd.)

Plants are the priceless sources of bioactive natural compounds. Medicinal plants have been used since time immemorial in both developing and developed countries. Flemingia macrophylla (Willd.) is widely used as a hereditary medicines. The current study was designed to observe the analgesic, antidiarrheal and CNS depressant activities of methanol extract of the leaves of F. macrophylla (MEFM) and its petroleum ether (PEFM), dichloromethane (DFM), chloroform (CFM) and aqueous (AFM) fractions. The analgesic activities was assessed by acetic acid induced writhing method at doses 200- and 400- mg/kg body weight. The CNS-depressant effect was assayed by phenobarbitone sodium-induced sleeping time test. The anti-diarrheal activity of the extract was evaluated using castor oil-induced diarrhea in mice. The crude extract displayed significant peripheral analgesic activity at both test doses with 56.72- to 59.70-% inhibition of writhing responses, respectively. In CNS-depressant test, the extract revealed its activity in a dose dependent manner. In screening for antidiarrheal activity, the extract exhibited 20.83 and 41.67 % inhibition of defecation at 200- and 400-mg/kg bw, respectively whereas the standard loperamide (50 mg/kg bw) displayed 70.83 % inhibition of defecation. Bangladesh Pharmaceutical Journal 23(2): 141-145, 2020

Multiple Pathway-Mediated Gut-Modulatory Effects of Maerua subcordata (Gilg) DeWolf.

BackgroundGastrointestinal disorders are often poorly managed, especially in developing countries, where there are limited resources and therapeutic options. Despite the rich diversity of medicinal plants that offer effective treatment options with fewer side effects, studies that provide scientific verification are lacking. Maerua subcordata (Gilg) DeWolf is among the plants claimed to have wide traditional medicine, use, including as a remedy against gastrointestinal problems. Therefore, this work aimed to evaluate the gut-modulatory effects of a crude leaf extract of M. subcordata (MSL.Cr), as well as its possible mechanism of action.MethodsA castor oil (10 mL/kg)–induced diarrheal mouse model was used to evaluate the antidiarrheal effect of MSL.Cr, and the spasmodic/antispasmodic effect of the extract was assessed using isolated rabbit jejunum with and without addition of standard cholinergic agonists/antagonists to predict the possible mechanism of action.ResultsMSL.Cr exhibited 40% and 80% protection against castor oil–induced diarrhea in mice at doses of 500 and 1,000 mg/kg, respectively. In isolated rabbit jejunum, the extract increased spontaneous contractions at low doses (0.01–0.1 mg/mL), and was sensitive to atropine, whereas it showed complete inhibition at higher doses (0.3–1 mg/mL). It was shown that the relaxant effect was possibly mediated by the involvement of phosphodiesterase-enzyme inhibition and K+-channel activation. The extract potentiated the control concentration–response curve of carbachol, shifting it to the left, similarly to the control drug papaverine. The potassium-channel opening–like activity of MSL.Cr was possibly mediated by the involvement of aspecific K+-channels inhibition, since tetraethylammonium, anunselective antagonist of K+ channels, significantly reversed its inhibitory effect.ConclusionThis study showed that the M. subcordata leaf extract demonstrated gut-modulatory effects, possibly mediated by a combination of muscarinic-receptor stimulation, phosphodiesterase inhibition, and aspecific K+-channel activation.

In Silico, Ex Vivo and In Vivo Studies of Roflumilast as a Potential Antidiarrheal and Antispasmodic agent: Inhibition of the PDE-4 Enzyme and Voltage-gated Ca++ ion Channels.

The aim of the present study was to evaluate the possible gut inhibitory role of the phosphodiesterase (PDE) inhibitor roflumilast. Increasing doses of roflumilast were tested against castor oil-induced diarrhea in mice, whereas the pharmacodynamics of the same effect was determined in isolated rabbit jejunum tissues. For in silico analysis, the identified PDE protein was docked with roflumilast and papaverine using the Autodock vina program from the PyRx virtual screening tool. Roflumilast protected against diarrhea significantly at 0.5 and 1.5 mg/kg doses, with 40% and 80% protection. Ex vivo findings from jejunum tissues show that roflumilast possesses an antispasmodic effect by inhibiting spontaneous contractions in a concentration-dependent manner. Roflumilast reversed carbachol (CCh, 1 µM)-mediated and potassium (K+, 80 mM)-mediated contractile responses with comparable efficacies but different potencies. The observed potency against K+ was significantly higher in comparison to CCh, similar to verapamil. Experiments were extended to further confirm the inhibitory effect on Ca++ channels. Interestingly, roflumilast deflected Ca++ concentration–response curves (CRCs) to the right with suppression of the maximum peak at both tested doses (0.001-0.003 mg/mL), similar to verapamil. The PDE-inhibitory effect was authenticated when pre-incubation of jejunum tissues with roflumilast (0.03-0.1 mg/mL) produced a leftward deflection of isoprenaline-mediated inhibitory CRCs and increased the tissue level of cAMP, similar to papaverine. This idea was further strengthened by molecular docking studies, where roflumilast exhibited a better binding affinity (-9.4 kcal/mol) with the PDE protein than the standard papaverine (-8.3 kcal/mol). In conclusion, inhibition of Ca++ channels and the PDE-4 enzyme explains the pharmacodynamics of the gut inhibitory effect of roflumilast.

Ellagic Acid Activated PPAR Signaling Pathway to Protect Ileums Against Castor Oil-Induced Diarrhea in Mice: Application of Transcriptome Analysis in Drug Screening.

BackgroundAcute diarrhea is still a common and serious disease. The causes of acute diarrhea are very complicated. Therefore, we need to find a medicine to control diarrhea symptoms, save time for diagnosis of pathogens, and prevent drug abuse. Ellagic acid (EA), a natural polyphenol drug, has anti-diarrhea effects. However, the action mechanisms of EA for non-specific diarrhea have not been characterized.Materials and MethodsTo study the mechanisms of EA, mice were divided into four groups. Group C were intraperitoneally injected with 0.1 ml physiological saline and orally given 0.2 ml physiological saline, and then after experiment began 0.5 h, orally administered 0.3 ml physiological saline. Group D were intraperitoneally injected with 0.1 ml physiological saline and orally given 0.2 ml castor oil, and then after experiment began 0.5 h, orally administered 0.3 ml physiological saline. Group E were intraperitoneally injected with 0.1 ml physiological saline and orally given 0.2 ml castor oil, and then after experiment began 0.5 h, orally administered 0.3 ml EA (10 mg/ml). Group V were intraperitoneally injected with 0.1ml GW9662 (1m g/ml) and orally given 0.2 ml castor oil, and then after experiment began 0.5 h, orally administered 0.3 ml EA (10 mg/ml). Transcriptome were performed on ileum tissues of mice in group D and E. Histological examination and qRT-PCR were performed on ileum tissues of mice in group C, D, E, and V.ResultsWe found that a total of 273 differentially expressed genes (DEGs) were obtained, including 160 up-regulated DEGs and 113 down-regulated DEGs. The DEGs were enriched in 458 Gene Ontology (GO) terms and 15 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, respectively. The peroxisome proliferator activated receptor (PPAR) signaling pathway was the most significantly enriched in KEGG pathways. We used the PPAR-specific antagonist GW9662 to validate the anti-diarrhea and anti-inflammatory effect of EA in group V compared with group E. Conclusively, EA protected ileums against castor oil-induced inflammation and diarrhea by activating the PPAR signaling pathway and a method was used to study the mechanism of EA.