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. 2015 Dec 30:10:93-105.
doi: 10.2147/DDDT.S91247. eCollection 2016.

The gastroprotective effects of hydroalcoholic extract of Monolluma quadrangula against ethanol-induced gastric mucosal injuries in Sprague Dawley rats

Ibrahim Abdel Aziz Ibrahim  1 , Mahmood Ameen Abdulla  2 , Maryam Hajrezaie  2 , Ammar Bader  3 , Naiyer Shahzad  1 , Saeed S Al-Ghamdi  1 , Ahmad S Gushash  4 , Mohadeseh Hasanpourghadi  5
Affiliations

The gastroprotective effects of hydroalcoholic extract of Monolluma quadrangula against ethanol-induced gastric mucosal injuries in Sprague Dawley rats

Ibrahim Abdel Aziz Ibrahim et al. Drug Des Devel Ther. .

Retraction in

Abstract

Monolluma quadrangula (Forssk.) Plowes is used in Saudi traditional medicines to treat gastric ulcers. The hydroalcoholic extract of M. quadrangula (MHAE) was used in an in vivo model to investigate its gastroprotective effects against ethanol-induced acute gastric lesions in rats. Five groups of Sprague Dawley rats were used. The first group was treated with 10% Tween 20 as a control. The other four groups included rats treated with absolute ethanol (5 mL/kg) to induce an ulcer, rats treated with 20 mg/kg omeprazole as a reference drug, and rats treated with 150 or 300 mg/kg MHAE. One hour later, the rats were administered absolute ethanol (5 mL/kg) orally. Animals fed with MHAE exhibited a significantly increased pH, gastric wall mucus, and flattening of the gastric mucosa, as well as a decreased area of gastric mucosal damage. Histology confirmed the results; extensive destruction of the gastric mucosa was observed in the ulcer control group, and the lesions penetrated deep into the gastric mucosa with leukocyte infiltration of the submucosal layer and edema. However, gastric protection was observed in the rats pre-fed with plant extracts. Periodic acid-Schiff staining of the gastric wall revealed a remarkably intensive uptake of magenta color in the experimental rats pretreated with MHAE compared to the ulcer control group. Immunohistochemistry staining revealed an upregulation of the Hsp70 protein and a downregulation of the Bax protein in rats pretreated with MHAE compared with the control rats. Gastric homogenate showed significantly increased catalase and superoxide dismutase, and the level of malondialdehyde (MDA) was reduced in the rats pretreated with MHAE compared to the control group. In conclusion, MHAE exhibited a gastroprotective effect against ethanol-induced gastric mucosal injury in rats. The mechanism of this gastroprotection included an increase in pH and gastric wall mucus, an increase in endogenous enzymes, and a decrease in the level of MDA. Furthermore, protection was given through the upregulation of Hsp70 and the downregulation of Bax proteins.

Keywords: Hsp70; Monolluma quadrangula; catalase; gastric ulcer; gastroprotective; malondialdehyde; superoxide dismutase.

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Figures

Figure 1
Figure 1
Histological investigation of acute toxicity analysis. Note: Effect of MHAE on histological sections of the liver and kidneys in an acute toxicity test (H&E stain). Abbreviations: MHAE, hydroalcoholic extract of Monolluma quadrangula; H&E, hematoxylin and eosin.
Figure 2
Figure 2
Gastric mucosal macroscopic evaluation. Notes: Effect of MHAE on the macroscopic appearance of the gastric mucosa in the ethanol-induced gastric mucosal injuries in rats. (A) The normal control group showed no injury to the gastric mucosa. (B) The ulcer control group showed severe hemorrhagic bands in the gastric mucosa. (C) The omeprazole group revealed mild hemorrhagic injuries and flattening of the gastric mucosa. (D) Rats pretreated with 150 mg/kg MHAE showed mild-to-moderate gastric mucosal injuries and flattening. (E) Rats pretreated with 300 mg/kg MHAE showed mild mucosal injuries and flattening. Abbreviation: MHAE, hydroalcoholic extract of Monolluma quadrangula.
Figure 3
Figure 3
H&E evaluation of the gastric mucosa. Notes: MHAE effect on the gastric mucosa histology in ethanol-induced gastric mucosal injury in rats. (A) The normal control group. (B) The ulcer control group exhibited a remarkably severe disruption to the gastric mucosa, which deeply penetrated into the mucosa with extensive edema and leukocyte infiltration of the submucosal layer. (C) The omeprazole group shows comparably mild disruption of gastric mucosa with mild edema and leukocytes infiltration of the submucosal layer. (D) Rats pretreated with 150 mg/kg showed mild-to-moderate gastric mucosa disruption with edema and leukocyte infiltration of the submucosal layer. (E) Rats pretreated with 300 mg/kg displayed mild disruption of the gastric mucosa with edema and leukocyte infiltration of the submucosal layer (H&E stain). The arrows show the H&E evaluation of the gastric mucosa. Abbreviations: H&E, hematoxylin and eosin; MHAE, hydroalcoholic extract of Monolluma quadrangula.
Figure 4
Figure 4
PAS staining evaluation. Notes: MHAE effect on gastric mucosal glycoprotein – PAS staining in ethanol-induced gastric hemorrhagic injuries in rats. (A) The normal control group. (B) The ulcer control group. (C) Rats pretreated with 20 mg/kg omeprazole. (D) Rats pretreated with 150 mg/kg MHAE. (E) Rats pretreated with 300 mg/kg MHAE. The arrows show the gastric mucosal glycoprotein production. Abbreviations: PAS, periodic acid–Schiff; MHAE, hydroalcoholic extract of Monolluma quadrangula.
Figure 5
Figure 5
Evaluation of Hsp70 protein expression in gastric tissue. Notes: The effect of MHAE on the expression of Hsp70 in ethanol-induced gastric mucosal injury in rats. (A) The normal control group. (B) The ulcer control group. (C) Rats pretreated with 20 mg/kg omeprazole. (D) Rats pretreated with 150 mg/kg MHAE. (E) Rats pretreated with 300 mg/kg MHAE (Hsp70 staining). The arrows show the level of Hsp70 protein expression. Abbreviation: MHAE, hydroalcoholic extract of Monolluma quadrangula.
Figure 6
Figure 6
Evaluation of Bax protein expression in gastric tissue. Notes: Effect of MHAE on the expression of Bax in ethanol-induced gastric mucosal injury in rats. (A) The normal control group. (B) The ulcer control group. (C) Rats pretreated with 20 mg/kg omeprazole. (D) Rats pretreated with 150 mg/kg MHAE. (E) Rats pretreated with 300 mg/kg MHAE (Bax staining). The arrows show the level of Bax protein expression. Abbreviation: MHAE, hydroalcoholic extract of Monolluma quadrangula.
Figure 7
Figure 7
Western blot of Hsp70 and Bax. Notes: (A) Western blot of (a) the normal control group, (b) ulcer control group. (c) rats pretreated with 20 mg/kg omeprazole, (d) rats pretreated with 150 mg/kg MHAE, and (e) rats pretreated with 300 mg/kg MHAE. (B) Relative intensity of each protein which was normalized with β-actin’s intensity. Data were mean ± SD of two independent experiments (P<0.05). Abbreviations: MHAE, hydroalcoholic extract of Monolluma quadrangular; SD, standard deviation.
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