Gastroprotective Effects of Plants Extracts on Gastric Mucosal Injury in Experimental Sprague-Dawley Rats

Abstract

Rubus crataegifolius (black raspberry, RF), Ulmus macrocarpa (elm, UL), and Gardenia jasminoides (cape jasmine, GJ) are well known for hundreds of years as folk medicines in China and Korea to treat various gastrointestinal disturbance. The present study evaluated the gastroprotective effects of these plants either single or in combination against HCl/EtOH-induced gastritis and indomethacin-induced ulcer in rat model. Stomach ulcer was induced by oral ingestions of HCl/EtOH or indomethacin. Treatment with RF, UL, and GJ separately or in combination was done 1 h before ulcer induction. On HCl/EtOH-induced gastritis RF, UL, and GJ at a dose of 150 mg/kg showed comparable antigastritis effect (less than 50% inhibition) with lesion index of 94.97±8.05, 108.48±11.51, and 79.10±9.77 mm compared to cimetidine (45.33±23.73 mm). However, the combination of RF, UL, and GJ at a dose of 150 mg/kg with a ratio of 50:50:50 showed remarkable antigastritis effect with 77% inhibition. The observed lesion index at a ratio of 50:50:50 was 23.34±9.11 mm similar to cimetidine (18.88±19.88 mm). On indomethacin-induced ulcer, RF and GJ showed 38.28% and 51.8% inhibition whereas UL showed around 17.73% inhibition at 150 mg/kg. Combination of RF, UL, and GJ at 150 mg/kg showed strong antigastritis effect with 83.71% inhibition. These findings suggest strong gastroprotective effect of combined extract. In addition, these plants showed significant antioxidant activity in DPPH scavenging assay and antilipid peroxidation activity. Combination of black raspberry, elm, and cape jasmine might be a significant systemic gastroprotective agent that could be utilized for the treatment and/or protection of gastritis and gastric ulcer.

Figure 1

HPLC-DAD chromatogram for standardization of sample. (a) ellagic acid standard at 254 nm, (b) Rubus crataegifolius extracts, (c) geniposide standard at 254 nm, (d) Gardenia jasminoides extracts, (e) catechin-7-O-ß-D-apiofuranoside at 280 nm, and (f) Ulmus macrocarpa extracts. The contents were standardized to contain 14.2 mg/g, 15.6 mg/g, and 30.5 mg/g, respectively.

Figure 2

Inhibitory effects of plants extract on HCl-EtOH-induced gastritis. (a) HCl-EtOH; (b) Cimetidine (150 mg/kg); (c) RF (150 mg/kg); (d) RF (300 mg/kg); (e) UL (150 mg/kg); (f) UL (300 mg/kg); (g) GJ (150 mg/kg); (h) GJ (300 mg/kg). n=7 for each group. The sum of erosion length (lesion index) per rat was determined using UTHSCSA Image Tool 3.00. Each value represents the mean ±SEM. ∗ p<0.05, ∗∗ p<0.01 vs. control rat. % inhibition of gastritis formation of each sample was determined.

Figure 3

Inhibitory effect of combined plant extracts on HCl-EtOH-induced gastritis. (a) HCl-EtOH, (b) Cimetidine (100 mg/kg), (c) RF combined UL (75+75 mg/kg), (d) RF combined GJ (75+75 mg/kg); (e) RF combined UL and GJ (50+50+50 mg/kg), and (f) RF combined UL and GJ (75+25+50 mg/kg). n=7 for each group. Gastric index was determined using UTHSCSA Image Tool 3.00. Each value represents the mean ±SEM. ∗ p<0.05, ∗∗ p<0.01 vs. control rat. % inhibition of gastritis formation of the combined samples was determined.

Figure 4

Inhibitory effects of plants extract on indomethacin-induced gastric ulcer. (a) Indomethacin treated group, (b) Cimetidine (100 mg/kg), (c) RF (150 mg/kg), (d) RF (300 mg/kg), (e) UL (150 mg/kg), (f) UL (300 mg/kg), (g) GJ (150 mg/kg), and (h) GJ (300 mg/kg). n=7 for each group. Gastric lesion index was determined using UTHSCSA Image Tool 3.00. Each value represents the mean ±SEM. ∗ p<0.05, ∗∗ p<0.01 vs. control rat. % inhibition of gastric value formation of each sample was determined.

Figure 5

Inhibitory effect of combined plant extracts on indomethacin-induced gastric ulcer. (a) Indomethacin, (b) Cimetidine (100 mg/kg), (c) RF combined UL (75+75 mg/kg), (d) RF combined GJ (75+75 mg/kg), (e) RF combined UL and GJ (50+50+50 mg/kg), and (f) RF combined UL and GJ (75+25+50 mg/kg). n=7 for each group. Gastric index was determined using UTHSCSA Image Tool 3.00. Each value represents the mean ±SEM. ∗ p<0.05, ∗∗ p<0.01 vs. control rat.

Figure 6

Effect of RF+UL+GJ plants extract on histological evaluation in HCl/ethanol and indomethacin-induced ulcer model. Section of hematoxylin and eosin (HE) staining of HCl/ethanol 5(a) and IMC-induced 5(b) gastric mucosa. Microscopic appearance of lesions induced by HCl/ethanol 5(a)(B) and IMC 5(b)(B) and lesion pretreated with 150 mg/kg of RUG-complex 5(a)(C) and 5(b)(C). Pathophysiological examination of the tissue sections was performed under light microscopy with 200x magnification.

Figure 7

Effect of RF+UL+GJ plants extract on lipid peroxidations. Effect of RF+UL+GJ complex on TBARS content on in vitro mouse stomach homogenate (Figure 6(a)). n=7 for each group. Each value represents the mean ±SEM. ∗ p<0.05, ∗∗ p<0.01 vs. control (Dunnett's multirange test). Effect of RF+UL+GJ complex in mice on HCl/ethanol-induced gastric damage (Figure 6(b)). n=7 for each group. 30, 90, 150, 300 mg/kg of the plants complex (at a dose of 1:1:1 ratio of RF, UL, and GJ) were used. Each value represents the mean ±SEM. ∗ p<0.05, ∗∗ p<0.01 vs. control (Dunnett's multirange test).