Anat Cell Biol.  2020 Sep;53(3):301-312. 10.5115/acb.20.055.

Gastroprotective effects of nebivolol and simvastatin against cold restraint stressinduced gastric ulcer in rats

• Kamar SS
• Latif NSA
• Elrefai MFM
• Amin SN

• KMID: 2507643
• DOI: http://doi.org/10.5115/acb.20.055

Abstract

Gastric ulcer is one of the most serious diseases. Nebivolol (Neb), a β1-blocker, exhibits vasodilator and antioxidative properties, simvastatin (Sim) antihyperlipidemic drug, exhibits anti-oxidative, anti-inflammatory properties and promote endogenous nitric oxide (NO) production. The aim of this study was to evaluate the gastroprotective effects of Neb and Sim against cold restraint stress (CRS)-induced gastric ulcer in rats. Rats were restrained, and maintained at 4°C for 3 hours. Animals were divided into six groups; control group, CRS group, and four treatment groups received ranitidine (Ran), Neb, Sim and both Neb and Sim. Treatments were given orally on a daily basis for 7 days prior to CRS. The gastroprotective effects of Neb and Sim were assessed biochemically by measuring variations in prostaglandins E2, NO, reduced glutathione and malondialdehyde, and functionally by estimating force of contractions of isolated rat fundus in the studied groups in response to acetylecholine stimulation and morphologically using hematoxylin and eosin staining, periodic acid Schiff ’s reaction and immunohistochemistry for cyclooxygenase 2 in gastric mucosa. CRS caused significant ulcerogenic effect. Alternatively, pretreatment with Ran, Neb, and Sim significantly corrected biochemical findings, pharmacological and histological studies.

Keyword

Gastric ulcer; Cold restraint stress; Ranitidine; Nebivolol; Simvastatin

Figure

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  Fig. 1 Force of contractions in the studied groups in response to acetylecholine stimulation. CRS, cold restraint stress; Neb, nebivolol; Ran, ranitidine; Sim, simvastatin.

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  Fig. 2 Comparison of Mean±SD of contraction force generated by acetyle choline stimulation in different studied groups. CRS, cold restraint stress; Neb, nebivolol; Ran, ranitidine; Sim, simvastatin. *Significant compared to Control group at P-value≤0.05; #Significant compared to CRS group at P-value≤0.05; @Significant compared to CRS+Ran group at P-value≤0.05; $Significant compared to CRS+Sim group at P-value≤0.05; &Significant compared to CRS+Neb group at P-value≤0.05.

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  Fig. 3 Photomicrograph of H&E stained sections in gastric mucosa (×100) showing: (A) Control group: intact mucosa with surface mucus layer (arrowheads) covering the gastric pits, gastric glands. (B) CRS group: diffuse mucosal abrasions, hemorrhage (asterisk), hemosiderin precipitation (arrow), patches of sloughed mucosal epithelium (curved arrows) and disturbed glands. (C) Ran+CRS group: intact gastric mucosa with intact surface epithelium. (D) Sim+CRS group: intact gastric mucosa with mild congestion in the lamina propria (curved arrow). (E) Neb+CRS group: ameliorated mucosal erosion with slight disturbance in the surface and cell desquamation (curved arrows). (F) Sim+Neb+CRS group: intact gastric mucosa with surface mucus layer (arrowheads) covering the gastric pits. CRS, cold restraint stress; Neb, nebivolol; Ran, ranitidine; RS, cold restraint stress; Sim, simvastatin. Scale bars=100 µm (A–F).

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  Fig. 4 Photomicrograph of PAS stained sections in gastric mucosa (×400) showing: (A) Control group: continuous strong PAS+ve reaction in the surface and neck region of the gastric glands. (B) CRS: mucosal abrasion with alternating absent to few PAS+ve reaction. (C) Ran+CRS: continuous strong PAS+ve reaction. (D) Sim+CRS: preserved PAS+ve reaction. (E) Neb+CRS: preserved PAS+ve reaction. (F) Sim+Neb+CRS: marked increase in the mucosal PAS+ve reaction. CRS, cold restraint stress; Neb, nebivolol; PAS, periodic acid Schiff’s; Ran, ranitidine; Sim, simvastatin. Scale bars=20 µm (A–F).

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  Fig. 5 Quantification analysis of the mean PAS+ve area %. CRS, cold restraint stress; Neb, nebivolol; PAS, periodic acid Schiff’s; Ran, ranitidine; Sim, simvastatin. *Significant decrease (P<0.05) when compared with control. #Significant increase (P<0.05) when compared with CRS.

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  Fig. 6 Photomicrograph of COX-2 immunostaining (×400) showing: (A) Control group: minimal COX-2+ve immunostaining in gastric mucosa. (B) CRS: minimal+ve immunostaining at the edge of the ulcer. (C) Ran+CRS: enhanced COX-2+ve immunostaining in the glandular epithelium and stromal cells. (D) Sim+CRS: moderate COX-2+ve immunostaining in the gastric mucosa. (E) Neb+CRS: moderate COX-2+ve immunostaining in the gastric mucosa. (F) Sim+Neb+CRS: marked increase COX-2+ve immunostaining in gastric mucosa. COX-2, cyclooxygenase 2; CRS, cold restraint stress; Neb, nebivolol; PAS, periodic acid Schiff’s; Ran, ranitidine; Sim, simvastatin. Scale bars=20 µm (A–F).

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  Fig. 7 Quantification analysis of the mean COX-2+ve immunostaining area %. COX-2, cyclooxygenase 2; CRS, cold restraint stress; Neb, nebivolol; Ran, ranitidine; Sim, simvastatin. *Significant increase (P<0.05) when compared with control and CRS groups. #Significant increase (P<0.05) when compared with Sim+CRS, @Significant increase (P<0.05) when compared with Neb+CRS groups.

Reference

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