Figure 1.
Chemical structure of Cu(BrHAP)2.
Table 1.
Elemental analysis and spectral characterization for the ligand and its metal complex.
Figure 2.
Histological sections in acute toxicity test (H & E staining, 20x).
Histological sections of liver (first row) and kidney (second row) in acute toxicity test. Rats treated with 5 mL/kg vehicle (10% Tween 20) (A and D). Rats treated with 500 mg/kg (5 mL/kg) the Copper (II) complex (B and E). Rats treated with 2000 g/kg (5 mL/kg) the Copper (II) complex (C and F). There is no significant differences in structures of liver and kidney between treated and control groups.
Figure 3.
Macroscopical appearance of the gastric mucosa in rats.
The negative control group (A) has no injury of gastric mucosa. The ulcer control group (B) shows sever injuries in the gastric mucosa. Absolute ethanol produced extensive visible hemorrhagic necrosis of gastric mucosa. The reference control group (omeprazole, 20 mg/kg) (C) shows mild injuries in the gastric mucosa comparing to the injuries seen in group 2. Rats received 10 mg/kg of the complex (D) has moderate injuries in the gastric mucosa. The extract reduces the formation of gastric lesions induced by absolute ethanol. Rats received 20 mg/kg of the complex (E) shows mild injuries in the gastric mucosa. Rats received 40 mg/kg of the complex (F) shows mild injuries in the gastric mucosa. Rats received 80 mg/kg of the complex (G) does not show any injuries of the gastric mucosa instead flattening of gastric mucosa is visible (white arrow). Black arrows point to the superficial hemorrhagic mucosal lesions.
Figure 4.
Effects of the complex on gastric ulcer area, inhibition percentage and alcian blue binding capacity.
Alcian blue binding capacity is defined as Gastric wall mucus (GWM). Groups 1 to 3 represent the negative control group, the ulcer control group and the reference control group (omeprazole, 20 mg/kg), respectively. The experimental groups received 10, 20, 40 and 80 mg/kg of the complex are presented as groups 4–7, respectively. All values are expressed as mean ± standard error mean. Mean difference is significant, in GWM, at the p<0.05 level (one-way between groups ANOVA with post-hoc analysis). * significant when compared with the group 2. # significant when compared with the group 3. Inhibition of gastric lesions (%) is indicated in brackets above the columns.
Figure 5.
Effect of the complex on glutathione (GSH), superoxide dismutase (SOD) and nitric oxide (NO) (A,B and C).
Groups 1 to 3 represent the negative control group, the ulcer control group and the reference control group (omeprazole, 20 mg/kg), respectively. The experimental groups received 10, 20, 40 and 80 mg/kg of the complex are presented as groups 4–7, respectively. All values are expressed as mean ± standard error mean. Mean difference is significant at the p<0.05 level (one-way between groups ANOVA with post-hoc analysis). * significant when compared with the group 2. # significant when compared with the group 3.
Figure 6.
Effects of the complex on malondialdehyde (MDA), prostaglandin E2 (PGE2) and protein concentration (A, B and C).
Groups 1 to 3 represent the negative control group, the ulcer control group and the reference control group (omeprazole, 20 mg/kg), respectively. The experimental groups received 10, 20, 40 and 80 mg/kg of the complex are presented as groups 4–7, respectively. All values are expressed as mean ± standard error mean. Mean difference is significant at the p<0.05 level (one-way between groups ANOVA with post-hoc analysis). * significant when compared with the group 2. # significant when compared with the group 3.
Figure 7.
Histological study of gastric mucosal damage in rats (20x).
In the negative control group 1 (A), no disruption to the surface epithelium is observed. The ulcer control group (B) has severe disruption to the surface epithelium (black arrows) and necrotic lesions penetrate deeply into mucosa and extensive edema of submucosal layer (yellow arrows) and leucocyte infiltration (blue arrows) are present. The reference control group (omeprazole, 20 mg/kg) (C), mild disruption of the surface epithelium mucosa is present but deep mucosal damage is absent. Reduction of submucosal edema and leucocytes infiltration is seen. Rats received 10 mg/kg of the complex (D) has moderate disruption of surface epithelium with submucosal edema and leucocytes infiltration of submucosal layer. Rats received 20 mg/kg of the complex (E) shows mild to moderate disruption of surface epithelium. In rats received 40 mg/kg of the complex (F), there is mild disruption to the surface epithelium. Reduction of submucosal edema and leucocytes infiltration of the submucosal layer are shown. Rats received 80 mg/kg of the complex (G) has no disruption to the surface epithelium.
Figure 8.
Effect of the complex on gastric tissue glycoprotein-PAS staining (20x).
The negative control group (A), the ulcer control group (B), the reference group (omeprazole, 20 mg/kg) (C), rats received 10 mg/kg of the complex (D), rats received 20 mg/kg of the complex (E), rats received 40 mg/kg of the complex (F), and rats received 80 mg/kg of the complex (G). Magenta color in the apical epithelial cells in the treated groups with the compound (groups 4–6) shows gradual increase in mucosal secretion of gastric glands. The intense secretion of mucus in gastric glands is demonstrated in group 7. The arrow points to the glycoprotein accumulation.
Figure 9.
Immunohistochemical analysis of expression of Hsp70 proteins (20x).
The negative control group (A), the ulcer control group (B), the reference group (omeprazole, 20 mg/kg) (C), rats received 10 mg/kg of the complex (D), rats received 20 mg/kg of the complex (E), rats received 40 mg/kg of the complex (F), and rats received 80 mg/kg of the complex (G). Immunohistochemistry staining of Hsp70 shows over-expression of Hsp70 protein in the experimental groups (D-G). The arrow points to the Hsp70 protein accumulation.
Figure 10.
Immunohistochemical analysis of expression of Bax proteins (20x).
The negative control group (A), the ulcer control group (B), the reference group (omeprazole, 20 mg/kg) (C), rats received 10 mg/kg of the complex (D), rats received 20 mg/kg of the complex (E), rats received 40 mg/kg of the complex (F), and rats received 80 mg/kg of the complex (G). Immunohistochemistry staining of Bax proteins shows down-expression of Bax protein in D-G. The arrow points to the Bax protein accumulation.