Spasmolytic Effect of Grewia asiatica Fruit Extract on Isolated Smooth Muscles is Mediated via Multiple Pathways

Background. Grewia asiatica Linn, or phalsa, is a commonly consumed fruit in Pakistan. The fruit is employed in the traditional medicine practice of Pakistan as a smooth muscle relaxant in different gastrointestinal (GI) and cardiovascular diseases. In this investigation, we show the antispasmodic and vasorelaxant actions of Grewia asiatica fruit extract. Methods. A 70% methanolic crude extract of the plant material was prepared (Ga.Cr). Different isolated GI tissue preparations and endothelium-intact aortas from rats were utilized to observe the pharmacological actions of the extract. Results. Ga.Cr, in increasing concentrations, inhibited the spontaneously contracting rabbit jejunum. In an effort to determine the mechanism of this relaxant action, contractions were induced in jejunum and ileum tissues with K+ (80 mM). Ga.Cr was able to only partially inhibit these induced contractions indicating that the mechanism might not be completely through a blockade of Ca2+ channels (CCB). When tested on low K+-(25 mM) sustained contractions, Ga.Cr cumulatively suppressed these contractions (0.1–10 mg/ml), indicating an opening of K+ channels (KCO) as the mechanism. Cromakalim, a standard KCO, was also more specific in blocking low K+-induced contractions. For the effect in aorta tissues, Ga.Cr suppressed the agonist-induced contractions from 0.3 mg/ml to 10 mg/ml. Upon challenge with L-NAME, a nitric oxide (NO) blocker, the extract response curve shifted right, indicating vasodilation was mediated via endothelial NO. Conclusion. This study shows that GI antispasmodic and vasodilator activities of Ga.Cr may be mediated via a KCO mechanism in the GI tract and through the release of NO from vascular endothelium.

Antidiarrhoeal potentials of methanol bark extract of Hymenocardia Acida Tul (Euphorbiaceae) in laboratory animals

Background The plant Hymenocardia acida (Euphorbiaceae) is utilized as herbal preparation against diarrhoea, dysentery and other diseases. We aimed to determine the antidiarrhoeal potentials of Hymenocardia acida (MEHA) stem bark in vivo and in vitro. Preliminary phytochemical contents, as well as the acute toxicity effect of the extract, were investigated based on standard experimental methods. The antidiarrhoeal properties of the MEHA at 150, 300 and 600 mg/kg were studied against diarrhoea induced by castor oil, intestinal fluid accumulation, as well as intestinal movement tests using distilled water (10 ml/kg) and loperamide/atropine sulphate as the control groups. Besides, the in vitro effects of the extract (8 × 10−2–640 × 10−2 mg/ml) on the rabbit jejunum and guinea-pig ileum were evaluated. Results Phytochemical screening showed alkaloids, glycoside, saponins, tannins, triterpenes, flavonoids and steroids in the MEHA. The median lethal dose (LD50) of the MEHA after oral administration was approximately greater than 2000 mg/kg. The MEHA declined the diarrhoea onset and remarkably decreased the number of watery stools in the group that received 300 and 600 mg/kg. It also elicited a remarkable and non-dose-dependent reduction in the intestinal fluid volume. At 1000 mg/kg, the MEHA significantly inhibited the charcoal movement. In addition, the MEHA (8 × 10−2–640 × 10−2 mg/ml) elicited a remarkable decrease in the contractility of the rabbit jejunum over time and relaxed the guinea pig ileum. Besides, it showed concentration-dependent attenuation of the acetylcholine and histamine-induced contraction. Conclusion The extract under investigation revealed promising antidiarrhoeal properties that justified its traditional claim for use against diarrhoea.

Anti-ulcer property of methanol fraction of Callichilia subsessilis leaf extract in albino rats

Peptic ulcer is one of the major causes of morbidity and mortality worldwide. It is treated with herbal preparations in developing countries. This study investigated the anti-ulcer property of methanol fraction of Callichilia subsessilis leaf in albino rats. Methanol extract prepared through the cold maceration method was partitioned into chloroform and methanol fraction using a separating funnel. The methanol fraction of Callichilia subsessilis (MFCS) was concentrated in vacuo using a rotary evaporator. The acute toxicity was determined using the brine shrimp lethality test and the up-and-down method at a dose limit of 2000 mg/kg. The anti-ulcer activity of MFCS was evaluated at the doses of 12.5, 25 and 50 mg/kg using indomethacin-and ethanol-induced ulcer models. Misoprostol was used as a reference standard. The ulcer score, index and severity were determined using standard methods. Isolated rabbit jejunum tissue in Tyrode’s solution was used to establish the possible mechanism of anti-ulcer activity of MFCS. The LC50 and LD50 of MFCS were greater than 10,000 ppm and 2,000 mg/kg respectively. The MFCS exhibited significant (p < 0.05) dose-dependent anti-ulcer activity in all the ulcer models used. The MFCS (25 mg/kg) produced 52% and 41.33% inhibition of ulcer index in the indomethacin- and ethanol-induced ulcer models respectively. In the isolated tissue model, MFCS caused significant (p < 0.05) relaxation of the rhythmic contraction of the isolated rabbit jejunum and partially inhibited acetylcholine- and histamine-induced contraction of the jejunal smooth muscles. In conclusion, MFCS exhibited anti-ulcer (antihistaminic and anticholinergic) effects. This study justified the use of Callichilia subsessilis leaves in traditional medicine as an anti-ulcer remedy.

Brief topical and intraluminal use of Carolina rinse solution does not attenuate experimental ischemia and reperfusion injury in rabbit jejunum

ABSTRACT Fifteen New Zealand adult rabbits were randomly allocated into three groups: Sham-operated (group A), Ischemia and Reperfusion (group B) and Carolina Rinse Solution (CRS) (group C). Groups B and C were subjected to one hour of ischemia and two hours of reperfusion. In group C, ten minutes before reperfusion, the bowel lumen was filled with CRS, and the segment immersed in CRS. Necrosis and loss of integrity of the villi were visible in groups B and C. Edema of the submucosa and circular muscle was observed in all groups. Hemorrhage was observed in different layers for groups B and C, but group C showed more severe hemorrhage in different layers during reperfusion. All groups showed polymorphonuclear leukocyte infiltration on the base of the mucosa, submucosa, and longitudinal muscle, in addition to polymorphonuclear leukocytes margination in the mucosal and submucosal vessels. Necrosis of enterocytes, muscles, crypts of Lieberkühn and myenteric plexus was observed in groups B and C during reperfusion. Topical and intraluminal Carolina Rinse Solution did not attenuate the effects of ischemia and reperfusion in the small intestine of rabbits.

Effect of Aqueous Ethanol Stem Extract of Entada africana Guill. Et Perrott. on Castor Oil and Magnesium Sulphate-Induced Diarrhoea Models in Mice

This study investigated the phytochemical, elemental analysis, acute toxicity study of aqueous ethanol stem bark extract of Entada africana, as well as its antidiarrhoeal activity in mice and its effects on isolated rabbit jejunum. The preliminary phytochemical screening revealed the presence of alkaloids, flavonoids, cardiac glycosides, phenols, saponins, steroids, tannins and terpenoids. Elemental analysis of the extract showed the presence of magnesium (Mg), manganese (Mn), iron (Fe), copper (Cu), lead (Pb), zinc (Zn) and sodium (Na) while acute toxicity study revealed intraperitoneal median lethal dose (LD50) values for the extract to be 774.6 mg/kg body weight. The antidiarrheal effect of the extract was studied using castor-oil and magnesium sulphate induced diarrhoeal models (dropping test) and gastrointestinal transit test in mice. The result showed that the extract produced a dose-dependent protection against diarrhoea induced by castor oil and magnesium sulfate, with the highest protection (80 and 100%), obtained at 100 and 200 mg/kg. The extract significantly (p≤0.01) reduced the small intestinal transit of charcoal meal in mice at all doses tested. The extract (0.4-3.2 mg/ml) produced a concentration dependent relaxation of the rabbit jejunum, and the effects were blocked by propranolol (0.04 and 0.64 μg/ml). The results of this study showed that the extract contain pharmacologically active substance with antidiarrhoeal properties mediated through inhibition of hyper secretion and reduced gastrointestinal motility. These properties may explain the rationale for use of it’s stem bark as antidiarrhoeal remedy in traditional medicine.

Antiplatelet Aggregation, Cardiotonic, Anti-Inflammatory, Antioxidant, and Calcium Channel Antagonistic Potentials of Nepeta ruderalis Buch

The objective of this study was to authenticate the ethnobotanical claims of the Nepeta ruderalis Buch.-Ham. (N. ruderalis) extract in the traditional system of medicine. Crude extract was prepared via a simple maceration process. DPPH free radical scavenging and carrageenan-induced rat paw edema models were used to monitor antioxidant and anti-inflammatory responses of the N. ruderalis extract. Furthermore, it was tested for antiplatelet aggregation, cardioprotective, and calcium channel antagonistic activities via standard documented protocols. The N. ruderalis extract exhibited 80.82% antioxidant activity (IC50=207.51±4.36 μg) while the anti-inflammatory response was significant (p<0.05 to p<0.01) at 50 mg/kg (45.58%) and 100 mg/kg (60.90%) doses. Moreover, it was found to inhibit platelet aggregation (IC50=1.06 and 0.91 mg/mL) and, in addition, to increase the force of contraction at the concentration of 3.0-10 mg/mL with a decrease in the heart rate on isolated paired atria (EC50=11.78 mg/mL). Relaxant activity was observed on the isolated rabbit jejunum (EC50=0.96 mg/mL) and trachea (EC50=0.89 mg/mL). However, in a cumulative way, an 80-millimolar potassium-induced contraction was evaluated (EC50=1.31 mg/mL). The N. ruderalis extract exhibited antioxidant, anti-inflammatory, platelet aggregating, cardiotonic, and calcium channel antagonistic activities, therefore proving scientifically its effectiveness in the traditional system of medicine.

Pharmacological and computational evaluation of fig for therapeutic potential in hyperactive gastrointestinal disorders

Background Ficus palmata (Fig), are distributed in different parts of the world, and are used in traditional medicine to treat various ailments including inflammation, tumor, epilepsy, jaundice, influenza and bacillary dysentery. The present study aimed to evaluate the antidiarrheal, antisecretary, antispasmodic, antiulcer and anti motility properties of Ficus palmata. Methods In-vivo, in-vitro and in-silico techniques were used to investigate various gastrointestinal effects of Ficus palmata. Antidiarrheal, antisecretary, antispasmodic, antiulcer, anti motility and molecular docking were performed using castor oil induced diarrhea and fluid accumulation, isolated tissue preparations, ethanol-HCl induced ulcer assay, charcoal meal transit time and Auto Doc Vina. Results Ficus palmata crude extract (Fp.Cr) exhibited protection against castor oil-induced diarrhea in mice and dose-dependently inhibited intestinal fluid secretions. Fp.Cr caused relaxation of spontaneous and K+ (80 Mm)-induced contractions in isolated rabbit jejunum preparations. It showed protective effect against gastric ulcers induced by ethanol-hydrochloric acid in rats. Fp.Cr reduced distance travelled by charcoal meal in the gastrointestinal transit model in mice. The plant constituents: psoralenoside and bergapten showed high binding affinities (E-value ≥ − 6.5 Kcal/mol) against histaminergic H1, calmodulin and voltage gated L-type calcium channels, while showed moderate affinities (E-value ≥7 Kcal/mol) against dopaminergic D2, adrenergic α1, muscranic M3, mu-opioid, whereas revealed lower affinities (E-value ≥9.5 Kcal/mol) vs. muscranic M1, histaminergic H2 and H+/K+ ATPase pump. Germanicol acetate and psoralene exhibited weak affinities against aforementioned targets. Conclusion This study reveals that Ficus palmata possesses anti-diarrheal, anti-secretory, anti-spasmodic, anti-motility and anti-ulcer activities. The various constituents reveal different binding affinities against target proteins, which mediate the gastrointestinal functions.

Preliminary Phytochemical Screening and Gastrointestinal Study on the Leaf Extract of Stachytarpheta angustifolia Mill Vahl (Verbenaceae) in Rabbit Jejunum

angustifolia (Verbenaceae) is mostly prescribed by the folkloric healers for various gastrointestinal disorders. This study was carried out to ascertain the gastrointestinal effect of the ethanol leaf extract and other various fractions (CHCl3, EtOAc, n- BuOH and residual aqueous) on rabbit Jejunum. The ethanol, n-butanol and residual aqueous of the extract exhibited dose concentration at (0.1, 0.2, 0.4 and 0.8 mg/ml) dependent contraction of the rabbit Jejunum which was blocked by atropine suggesting that the observed pharmacological actions was mediated through the muscarinic receptors. In contrast, chloroform and ethylacetate fraction of the leaf extract exhibit dose concentration dependent relaxation of the rabbit jejunum. Intreperitoneal LD50 of the extract in mice was found to be 295.8 mg/kg. Preliminary phytochemical screening of the leaf extract revealed the presence of carbohydrates, tannins, saponins, cardiac glycoside, sterols and terpenoids. The result indicated that, the plant extract possesses some pharmacological activity, hence justifying its use traditionally in alleviating gastrointestinal disorder.

Pharmacological Studies Pertaining to Smooth Muscle Relaxant, Platelet Aggregation Inhibitory and Hypotensive Effects of Ailanthus altissima

Objective. This in vitro and in vivo study was conducted to rationalize some of traditional medicinal uses of Ailanthus altissima in gastrointestinal, respiratory, and cardiovascular systems. Materials. Crude extract of Ailanthus altissima (Aa.Cr) and its fractions were prepared and utilized in in vitro and in vivo studies. For in vitro studies, Aa.Cr was investigated on isolated rabbit jejunum, isolated tracheal strip, and isolated aorta of rat suspended in tissue organ bath. Platelet rich and platelet poor plasma were used to study platelet aggregation inhibitory activity. In vivo antidiarrheal effect of Aa.Cr was investigated on balb/c mice pretreated with castor oil to induce diarrhea and SD rats were used to study hypotensive activity. Results. Concentration dependent spasmolytic effects of Aa.Cr and its DCM fraction (Aa.DCM) were observed on spontaneous and spasmogen induced contractions in jejunum isolated from rabbit, but effect against high potassium (high-K+) induced contractions was more potent. Moreover Aa.Cr showed parallel shifting of calcium response curve to the right side. While its aqueous fraction (Aa.aq) caused spasmogenesis of isolated rabbit jejunum, this effect was blocked partially with prior administration of atropine (1μM). Concentration dependent protection against castor oil induced diarrhea was also observed. Relaxant effect was observed by the application of Aa.Cr and Aa.DCM against high-K+ and carbachol (CCh) induced contractions in tracheal strips isolated from SD rats, while Aa.Aq caused partial relaxation of high-K+ induced contractions, but no effect was observed against CCh induced contractions. Relaxation of rat aorta by the application of Aa.Cr and its fractions was also observed. Inhibition of force of contraction in rabbit atrium was also observed. Inhibition of platelet aggregation was observed against epinephrine and ADP induced aggregation. Conclusion. Keeping in view the observed results, it is concluded that smooth muscle relaxant, platelet aggregation inhibitory and hypotensive effect may be due to the blockage of calcium channels.