Achillea millefolium Essential Oil Mitigates Peptic Ulcer in Rats through Nrf2/HO-1 Pathway

Abstract

Extreme ethanol ingestion is associated with developing gastric ulcers. Achillea millefolium (yarrow) is one of the most commonly used herbs with numerous proven pharmacological actions. The goal of the hereby investigation is to explore the gastroprotective action of yarrow essential oil against ethanol-induced gastric ulcers and to reveal the unexplored mechanisms. Rats were distributed into five groups (n = 6); the control group administered 10% Tween 20, orally, for two weeks; the ethanol group administered absolute ethanol (5 mL/kg) to prompt gastric ulcer on the last day of the experiment. Yarrow essential oil 100 or 200 mg/kg + ethanol groups pretreated with yarrow oil (100 or 200 mg/kg, respectively), orally, for two weeks prior to gastric ulcer induction by absolute ethanol. Lanso + ethanol group administered 20 mg/kg lansoprazole, orally, for two weeks prior to gastric ulcer induction by ethanol. Results of the current study showed that ethanol caused several macroscopic and microscopic alterations, amplified lipid peroxidation, pro-inflammatory cytokines, and apoptotic markers, as well as diminished PGE₂, NO, and antioxidant enzyme activities. On the other hand, animals pretreated with yarrow essential oil exhibited fewer macroscopic and microscopic modifications, reduced ulcer surface, and increased Alcian blue binding capacity, pH, and pepsin activity. In addition, yarrow essential oil groups exhibited reduced pro-inflammatory cytokines, apoptotic markers, and MDA, restored the PGE₂ and NO levels, and recovered the antioxidant enzyme activities. Ethanol escalated Nrf2 and HO-1 expressions, whereas pretreatment of yarrow essential oil caused further intensification in Nrf2 and HO-1. To conclude, the current study suggested yarrow essential oil as a gastroprotective agent against ethanol-induced gastric lesions. This gastroprotective effect could be related to the antioxidant, anti-inflammatory, and anti-apoptotic actions of the essential oil through the instigation of the Nrf2/HO-1 pathway.

Keywords: Achillea millefolium; Nrf2/HO-1 pathway; anti-inflammatory; antioxidant; apoptosis.

Figure 1

The actions of YEO (100 and 200 mg/kg) administration in acute gastric mucosal injuries induced by ethanol in rats on (a) demonstrative images of the separated gastric and (b) gastric injury scores. All data are quantified as mean ± SE, (n = 6). (#) defines statistically significant related to the control, (*) signifies statistically significant related to the ethanol-induced gastric mucosal injuries, and (€) designates statistically significant related to YEO 200 + ethanol via one-way ANOVA afterward Tukey’s post hoc test (p < 0.05).

Figure 2

The actions of YEO (100 and 200 mg/kg) administration in acute gastric mucosal injuries induced by ethanol in rats on (a) histological assessment of the gastric tissue staining with H&E and the scores for (b) hemorrhagic injury, (c) epithelial cell loss, and (d) edema with leucocytes. Blue arrows show necrotic lesions, black arrowheads display disrupted surface epithelium and green arrowheads indicate leucocyte infiltration. (#) defines statistically significant related to the control, (*) signifies statistically significant related to the ethanol-induced gastric mucosal injuries, and (€) designates statistically significant related to YEO 200 + ethanol.

Figure 3

Effects of YEO (100 and 200 mg/kg) administration in acute gastric mucosal injuries induced by ethanol in rats on immunohistochemical analysis of (a) Nrf2 and HO-1 expressions, immunohistochemical scores for (b) Nrf2 and (c) HO1 and on the gene expression of (d) Nrf2 and (e) HO1. All data are quantified as mean ± SE, (n = 6). (#) defines statistically significant related to the control, (*) signifies statistically significant related to the ethanol-induced gastric mucosal injuries, and (€) designates statistically significant related to YEO 200 + ethanol via one-way ANOVA after Tukey’s post hoc test (p < 0.05).

Figure 4

Effects of YEO (100 and 200 mg/kg) administration in acute gastric injuries induced by ethanol on serum inflammatory cytokines markers including (a) IL-1β, (b) IL-6, and (c) TNF-α and on gastric mucosa defensive factors including (d) PEG2, (e) serum NO, and (f) gastric NO. All data are quantified as mean ± SE, (n = 6). (#) defines statistically significant related to the control, (*) signifies statistically significant related to the ethanol-induced gastric mucosal injuries, and (€) designates statistically significant related to YEO 200 + ethanol via one-way ANOVA after Tukey’s post hoc test (p < 0.05).

Figure 5

Effects of YEO (100 and 200 mg/kg) administration in acute gastric mucosal injuries induced by ethanol in rats on gastric and serum of (a,b) MDA, (c,d) GSH, (e,f) SOD and (g,h) catalase, respectively. All data are quantified as mean ± SE, (n = 6). (#) defines statistically significant related to the control, (*) signifies statistically significant related to the ethanol-induced gastric mucosal injuries, and (€) designates statistically significant related to YEO 200 + ethanol via one-way ANOVA afterward Tukey’s post hoc test (p < 0.05).

Figure 6

Effects of YEO (100 and 200 mg/kg) administration in acute gastric mucosal injuries induced via ethanol in rats on apoptotic indicators including (a) caspase-3, (b) caspase-3 and 9, and the gene expression of (c) Bax and (d) Bcl2. All data are quantified as mean ± SE, (n = 6). (#) defines statistically significant related to the control, (*) signifies statistically significant related to the ethanol-induced gastric mucosal injuries, and (€) designates statistically significant related to YEO 200 + ethanol via one-way ANOVA afterwards Tukey’s post hoc test (p < 0.05).