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. 2019 Jun 5;10(4):345-353.
doi: 10.1016/j.jtcme.2019.06.001. eCollection 2020 Jul.

Gastroprotective and anti-inflammatory effects of Prunus cerasus phytochemicals and their possible mechanisms of action

Karim Raafat  1 , Nada El-Darra  2 , Fatima A Saleh  3
Affiliations

Gastroprotective and anti-inflammatory effects of Prunus cerasus phytochemicals and their possible mechanisms of action

Karim Raafat et al. J Tradit Complement Med. .

Abstract

Prunus cerasus (P. cerasus) is an alternative-medicine used traditionally for amelioration of chronic-ailments marked by elevation in oxidative-stress like neuropathy. The oxidative-stress control was reported to ameliorate the inflammatory-process. This study aimed to phytochemically-investigate P. cerasus most-active phytochemicals utilizing in-vivo biological models to explore their gastroprotective, anti-inflammatory, and antinociceptive potentials and their possible mechanisms of action. Sonication with EtAc was used to extract P. cerasus fruit (Scf), and seed (Scs). The phytochemical-investigation of Scf was performed by RP-HPLC, while that of Scs was explored utilizing GC-FID. A bio-guided-fraction and isolation method was done utilizing column-chromatography, and have shown that cyanidin-3-glucoside (Cy3G) was the most-active constituent in Scf, while linoleic-acid (LA) was the most-active constituent in Scs. Scf, Scs, Cy3G, and LA significantly (p ˂ 0.05) protected the gastric-mucosa against HCl/EtOH-induced gastric-lesions. Scs (200 mg/kg) has shown the most gastroprotective-potentials, and had comparable-results to ranitidine (50 mg/kg). Scf, Scs, Cy3G, and LA have shown significant anti-inflammatory and antinociceptive potentials against carrageenan induced-edema and nociceptive-pain, respectively, where Scs (200 mg/kg) has shown the most anti-inflammatory and antinociceptive potentials, and had comparable results to ibuprofen (100 mg/kg). Scf, Scs, Cy3G, and LA have counter-acted carrageenan-induced oxidative-stress markers, with increased serum-catalase and reduced-glutathione levels, and decreased lipid-peroxidation. Histopathological-studies demonstrated gastroprotective potentials, regeneration and improvement of the spleen-structural architecture when treated with highest doses of Scs and Scf. The reduction of the pro-inflammatory TNF-alpha and IL-6, and elevation the anti-inflammatory factor IL-10 levels, spleen regenerative-capacity and oxidative-stress amelioration might be the main-mechanism responsible for P. cerasus anti-inflammatory potentials. P. cerasus appears to aid in ameliorating the inflammatory process, and reducing pain-thresholds while preserving the stomach.

Keywords: Anti-inflammatory; Antinociceptive effects; Cy3G, Cyanidin 3-glucoside; EtAc, Ethyl acetate; EtOH, ethanol; FID, flame-ionization detector; GSH, reduced glutathione; Gastroprotective; H and E staining, Hematoxylin and Eosin staining; HAc, acetic acid; IL-10, Interleukin 10; IL-6, Interleukin 6; Ib, Ibuprofen; LA, Linoleic acid; LPO, lipid peroxidation; MeOH, methanol; NSAIDs, Non-steroidal anti-inflammatory drugs; Oxidative stress; P. cerasus, Prunus cerasus; PWT, paw withdrawal threshold; Prunus cerasus; Scf, sour cherry fruit ethyl acetate extract; Scs, sour cherry seed ethyl acetate extract; TBARS, Thiobarbituric acid reactive substances; TNF-alpha, Tumor necrosis factor alpha; VEH, vehicle control; e, edema; er, erosions; h, hemorrhage; ic, infiltration of inflammatory cell in the sub-mucosa; mu, mucosa; sm, sub-mucosa.

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Figures

Image 1
Graphical abstract
Fig. 1
Fig. 1
GC-FID analysis showing the composition of fatty acids (mole percentage) of Sour cherry seed EtAc extract (Scs). Values are means of triplicate determinations ± S.E.M. Values with different letters are significantly different (p < 0.05).
Fig. 2
Fig. 2
Gastroprotective activities against HCl/EtOH-induced gastric lesion in mice: (A) Normal Control (Untreated) (B) Vehicle Control (C) Ranitidine-HCL (D) Sour Cherry Fruit 100 mg/Kg-HCL (E) Sour Cherry Fruit 150 mg/Kg-HCL (F) Sour Cherry Fruit 200 mg/Kg-HCL (G) Sour Cherry Seed 100 mg/Kg-HCL (H) Sour Cherry Seed 150 mg/Kg-HCL (I) Sour Cherry Seed 200 mg/Kg-HCL (J) Cyanidin-3-O glycoside (Cy3G) 10 mg/Kg-HCl (K) Cy3G 15 mg/Kg-HCl (L) Cy3G 20 mg/Kg-HCl (M) Linoleic acid (LA) 10 mg/Kg-HCl (N) LA 15 mg/Kg-Hcl (O) LA 20 mg/Kg-HCl.
Fig. 3
Fig. 3
Histological alterations in stomach sections of HCl/EtOH-induced gastric lesion in mice -induced gastric lesions in mice: (A) Microphotographs of normal control (NC) mice showing normal histological appearance of the mu (mucosa) and sm (sub-mucosa). (B) Stomach from mice treated with ranitidine-HCl, showing normal histological appearance of mu and sm. (C) Microphotographs of vehicle treated (vehicle control) HCl/EtOH, showing edema (e), severe-erosion (er), and hemorrhage (h) in the mu, and ic (infiltration of inflammatory-cells in the sm). (D) Stomach from mice treated with Scf 200 mg/kg 60 min before HCl/EtOH administration, showing mild e, and absence of er, h and ic. (E) Stomach from mice treated with Scs 200 mg/kg before HCl/EtOH administration showing normal histology of the mu and sm. (F) Microphotographs of mice treated with Cy3G 20 mg/kg before HCl/EtOH administration, showing mild e and er, and absence of h and ic. (G) Stomach from mice treated with LA 20 mg/kg before HCl/EtOH administration showing normal histology of the mu and sm.
Fig. 4
Fig. 4
Antinociceptive effects of Scf, Scs, Cy3G and LA against acute carrageenan-induced inflammatory-pain (mean ± SEM). Carrageenan (100 μL, 1%) was injected intraplantarly 2 h prior the pain threshold measurement by the paw pressure test. Sour cherry fruit EtAc extract (Scf), Sour cherry seed EtAc extract (Scs), Cyanidin 3-O-Glycoside (Cy3G), and Linoleic acid (LA) were administered 30 min before the test. Ibuprofen 100 mg/kg (Ib 100) was used as a positive control. “Normal” designates normal mice. “*” means  P < 0 .05, n = 7 compared with vehicle control (VEH).
Fig. 5
Fig. 5
The anti-inflammatory effects on mouse hind paw edema of: (A) Sour cherry Fruit EtAc extract (Scf) and sour cherry seed EtAc extract (Scs), and (B) cyanidin 3-glucoside (CY3G) and linoleic acid (LA). All doses were in mg/Kg. The actual edema volume increase was measured relative to that of standard drug ibuprofen 100 mg/kg. Values are presented as means ± SEM, n = 7. ∗ denotes significant difference from control value at P < 0.05.
Fig. 6
Fig. 6
Representative Hematoxylin and Eosin (H and E) stained sections of spleen-tissue from (A) Normal control (NC) (B) Vehicle control (CTRL) (C) Group administered 200 mg/kg Scs and (D) Group administered 200 mg/kg Scf. Top panel 4× magnification; Lower panel 10× magnification; red pulp (RP); white pulp (WP), central arteriole (arrow).
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