Synergistic Effect of Rubus crataegifolius and Ulmus macrocarpa Against Helicobacter pylori Clinical Isolates and Gastritis

Abstract

Helicobacter pylori is one of the most widespread infections involved in the pathogenesis of chronic gastritis, peptic ulcer, and gastric cancer. Hence, there is an urgent need to develop medications against H. pylori. This study aimed to evaluate synergistic effect of Rubus crataegifolius (RF) and Ulmus macrocarpa Hance (UL) against H. pylori. Antibacterial susceptibility of each extract either separately or in combination was studied against two H. pylori standard strains and 11 clinical isolates using agar dilution method. The effect of the extracts on H. pylori inoculated Balb/c mice model was also studied using single dosing (100 mg/kg each) approach. The MIC₅₀ of RF and UL were more than 100 and 200 µg/ml, respectively, against the tested strains. However, simultaneous treatment with RF and UL at 75 and 50 µg/ml, respectively, showed decreased viable cell number, MIC₇₀, and at 75 µg/ml each showed synergic effect with MIC₉₀. On H. pylori inoculated Balb/c mice model, RF and UL separately (100 mg/kg each) showed moderate anti-H. pylori effect, while simultaneous treatment of RF and UL with same dose showed significant synergistic anti-gastric effects in stomach. The results showed a significant synergistic effect of plants extract against H. pylori infection and eventually gastric mucosal damage. Our finding could be considered a valuable support in the treatment of H. pylori induced gastritis and may contribute to the development of new and safe combined herbal product as anti-H. pylori regimens.

Keywords: Helicobacter pylori; MIC; Rubus crataegifolius; Ulmus macrocarpa; gastritis.

Figure 1

HPLC-DAD chromatogram for standardization of sample. (A) Rubus crataegifolius extracts and standard ellagic acid (overlaid) at 254 nm, (B) Ulmus macrocarpa extracts and standard catechin-7-O-β-D-apiofuranoside (overlaid) at 280 nm.

Figure 2

Synergistic effects of plant extracts on the colonization of H. pylori. (A) Effects of plant extracts (Rubus crataegifolius, Ulmus macrocarpa) on the colonization of 13 (represented as alphabet) H. pylori strains determined by agar dilution method, (B) Percentage of inhibition of colonization of each extract represented as graph. RF combined with UL at 75 µg/ml each showed complete inhibition of H. pylori growth (MIC₉₀) on complex treated agar plates. CON indicated normal control and 75 indicated 75 µg/ml.

Figure 3

Histological analysis for H & E staining (magnification × 40). (A) H. pylori infected control mouse 4 weeks, (B) H. pylori infected control mouse 8 weeks, (C) Treatment with triple therapy (OAC), (D) Treatment with R. crataegifolius, (E) Treatment with U. macrocarpa, (F) Treatment with R. crataegifolius combined U. macrocarpa. Each plant extract suppresses immune cells against inflammation in mice stomach. Histological scoring of inflammation: normal (0), mild (1), moderate (2), and severe (3).