Pharmacological investigation of brucine anti-ulcer potential

Abstract

Gastric ulcer is one of the most common chronic gastrointestinal diseases characterized by a significant defect in the mucosal barrier. The current study has been conducted to evaluate the brucine anti-ulcer effect. Brucine has binding energy values ranging from -2.99 to -8.11 kcal/mol against chosen targets, according to in silico research. Brucine exhibits an inhibitory effect against Helicobacter pylori. In vivo findings revealed that brucine (3 mg/kg) showed effective results in healing ethanol-induced ulcer lesions of the gastric region in rats. Brucine showed an inhibitory effect against H⁺/K⁺-ATPase. Levels of glutathione, glutathione-s-transferase, and catalase were enhanced in the gastric rat tissue with the use of brucine, while a significant decrease in lipid peroxide levels was seen. Histopathological evaluation showed improvement in cellular architecture and a decrease in inflammatory indicators like cyclooxygenase, tumor necrosis factor, and nuclear factor kappa B expression, validated through immunohistochemistry, enzyme-linked immunosorbent assay, and Western blot techniques. In the reverse transcription-polymerase chain reaction, brucine decreased H⁺/K⁺-ATPase mRNA levels. This study reveals that brucine possesses stable binding affinities against selected targets. Brucine exhibits an anti-ulcer effect, mediated via anti-H. pylori, H⁺/K⁺-ATPase inhibition, and antioxidant and anti-inflammatory pathways.

Keywords: H+/K+-ATPase; anti-H. pylori; anti-inflammatory; anti-ulcer; antioxidant; brucine.

FIGURE 1

(A, B) represent 2D and 3D structures of brucine.

FIGURE 2

Gross appearance of gastric mucosa in rats: (A) pretreated with saline, (10 mL/kg). (B) Severe injuries are seen, extensive visible hemorrhagic necrosis of gastric mucosa was produced by absolute ethanol (1 mL/100g), (C, D) pretreated with brucine at doses of 1 and 3 mg/Kg, and (E) pretreated with omeprazole 20 mg/Kg.

FIGURE 3

Inhibitory effect of brucine and omeprazole against H+/K+-ATPase in ethanol-induced ulcer rat gastric tissues. Values expressed as mean ± SEM (n = 5). One-way ANOVA with post hoc Tukey’s test was applied. ^###p < 0.001 vs. saline group; ∗∗∗p < 0.001 vs. ethanol group.

FIGURE 4

Effect of brucine and omeprazole against glutathione sulfotransferases (GST), reduced glutathione (GSH), catalase, and lipid peroxidase (LPO) in ethanol-induced ulcer rat gastric tissues. Data presented as mean ± SEM (n = 5). One-way ANOVA with post hoc Tukey’s test. ^### p < 0.001 vs. saline group; ^∗∗∗ p < 0.001 vs. ethanol group.

FIGURE 5

Histopathological slides represent effect of brucine and omeprazole in ethanol- induced ulcer rats gastric tissues. Saline group showing normal histological features. Ethanol ulcer group showing marked histopathological deformities by loss of stomach architecture, vacuolization and cloudy swelling. Brucine and omeprazole treatment groups showing near normal architecture with mild to moderate deformities.

FIGURE 6

Slides represent the effect of brucine and omeprazole against the expressions of cyclooxygenase (COX-2), nuclear factor kappa B (p-NFκB), and tumor necrosis factor alpha (TNF-α) in ethanol-induced ulcer rat gastric tissues, using the immunohistochemical technique.

FIGURE 7

Effects of brucine and omeprazole against cyclooxygenase (COX-2), nuclear factor kappa B (p-NFκB), and tumor necrosis factor alpha (TNF-α) in ethanol-induced ulcer rat gastric tissues, using the immunohistochemical technique. Values expressed as mean ± SEM (n = 5). Data were analyzed by one-way ANOVA, followed by post hoc Tukey’s test. ^### p < 0.001 vs. saline group. ^∗∗∗ p < 0.001 vs. ethanol group.

FIGURE 8

Effects of brucine and omeprazole against nuclear factor kappa B (p-NFκB), prostaglandin (PGE₂), and tumor necrosis factor (TNF-α) levels in ethanol-induced ulcer rat gastric tissues, using the enzyme-linked immunosorbent assay technique. Values expressed as mean ± SEM (n = 5). One-way ANOVA with post hoc Tukey’s test. ^### p < 0.001 vs. saline group. *p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 vs. ethanol group.

FIGURE 9

Bands (A) and graphical (B) representation of effects of brucine and omeprazole against phosphorylated nuclear factor kappa B (p-NFκB) and tumor necrosis factor (TNF-α) expressions in ethanol-induced ulcer rat gastric tissues, using the Western blot technique. Values expressed as mean ± SEM (n = 5). One-way ANOVA with post hoc Tukey’s test. ^### p < 0.001 vs. saline group. ***p < 0.001 vs. ethanol group.

FIGURE 10

Effect of brucine and omeprazole against H⁺/K⁺-ATPase mRNA expression in ethanol-induced ulcer-treated rat gastric tissues, using the reverse transcription–polymerase chain reaction (RT-PCR) technique. One-way ANOVA followed by post hoc Tukey’s test. Values expressed as mean ± SEM (n = 5). ^### p < 0.001 vs. saline group; ∗∗∗p < 0.001 vs. ethanol group.