Effect of Rumex dentatus on Gastrointestinal Protection and Toxicology in Rodents via Investigating H+/K+-ATPase, Calcium Channels, and PDE Mediated Signaling

Abstract

This present study aims to delineate Rumex dentatus crude extract (Rd.Cr), n-Hexane, ethyl acetate, aqueous fractions (Rd.n-Hex, Rd.ETAC, and Rd.Aq), and emodin for antidiarrheal, antisecretory effects, anti-spasmodic, gastrointestinal transient time, anti-H. pylori, antiulcer effects, and toxicology. Plant extracts attributed dose-dependent protection against castor oil-induced diarrhea and dose-dependently inhibited intestinal fluid secretions in mice. They decreased the distance transverse by charcoal in the gastrointestinal transit model in rats. In rabbit jejunum preparations, it causes a concentration-dependent relaxation of both spontaneous and K+ (80 mM)-induced contraction, Rd.n-Hex and verapamil were relatively potent against K+-induced contractions and shifted the Ca2+ concentration-response curves (CRCs) to the right, Rd.Cr and Rd.ETAC shifted the isoprenaline-induced inhibitory CRCs to the left, showing potentiating effect similar to papaverine. Rd.n-Hex showed anti-H. pylori effect. Extracts and emodin also show an inhibitory effect against H+/K+-ATPase. Rumex dentatus showed a gastroprotective and antioxidant effect. Histopathological evaluation showed improvement in cellular architecture and decrease in the expression of inflammatory markers such as cyclooxygenase (COX2), tumor necrosis factor (TNF-α), and phosphorylated nuclear factor kappa B (p-NFƙB), validated through immunohistochemistry, ELISA, and western blot techniques. In RT-PCR, it decreases H+/K+-ATPase mRNA levels. Rumex dentatus was analyzed for certain safety aspects and exhibited a relative safety profile as no impairment was observed in kidneys, heart, liver, and brain further assisted by biochemical and hematological analysis. Docking studies revealed that emodin against H+/K+-ATPase pump and voltage gated L-type calcium channel showed E-value of −7.9 and −7.4 kcal/mol, respectively. MD simulations and molecular mechanics Poisson Boltzmann surface area and molecular mechanics Generalized Born surface area MMPBSA/GBSA findings are consistent with the in-vitro, in-vivo, and docking results. In conclusion, Rumex dentatus extracts and its phytoconstituent could be considered a potent antioxidant and anti-inflammatory drug candidates that possess anti-diarrheal, anti-secretary, antispasmodic, anti-H. pylori, and anti-ulcer potential. Toxicity studies were done according to OECD standards 425. It belongs to group 5 (LD50 > 2000 mg/kg), which suggests that it is in the lower toxicity class.

Pharmacological basis of bergapten in gastrointestinal diseases focusing on H+/K+ ATPase and voltage-gated calcium channel inhibition: A toxicological evaluation on vital organs

“…For charcoal meal transient time activity, bergapten in the small intestinal produced suppression of propulsion of charcoal marker in test doses just like standard drug atropine sulfate which has anticholinergic effect on GI transit leading to reduced motility ( Ansari et al, 2019 ). The reduced motility causes sufficient reabsorption of water and contents through the intestine which can also be correlated to antidiarrheal findings of bergapten in this study ( Qazi et al, 2022 ). Other functional regulators involved in GI motility include acetylcholine, histamine, prostaglandins and ionic exchanges involving K + and Ca ++ ions ( Sadraei et al, 2022 ).…”

“…To assess the effect of bergapten in gastrointestinal disorders such as castor oil-induced diarrhea, this study was design to measure the antidiarrheal effect of bergapten in rats similar to effect produced by loperamide, a standard drug while its possibly underlying mechanism were estimated through isolated tissue preparations also associated with reduction in gastric motility ( Qazi et al, 2022 ). Castor oil is responsible for increasing intestinal fluid as well as diarrheal effect through its active metabolite, that is, ricinoleic acid.…”

“…LPO (lipid peroxidation) values are measured in TBARS (thiobarbituric acid reactive substances) nmoles/min/mg protein. The end product of LPO is malondialdehyde, which is measured using a microplate reader and absorbance at 532 nm ( Qazi et al, 2022 ). For SOD (superoxide dismutase), an assay was performed according to Dengiz et al (2007 ).…”

“…The end product of LPO is malondialdehyde, which is measured using a microplate reader and absorbance at 532 nm (Qazi et al, 2022). For SOD (superoxide dismutase), an assay was performed according to Dengiz et al (2007) activity was estimated by inhibition of the aforementioned reaction and inhibition determined at 560 nm and expressed as millimole per minute per milligram tissue (mmol/min/mg/tissue).…”

Anti-diabetic Activity of Brucine in Streptozotocin-Induced Rats:In Silico,In Vitro, andIn VivoStudies

“…LPO values were calculated using TBARS nmoles/min/mg of proteins. 40 4.11. Hematoxylin and Eosin (H&E) Staining.…”