Saffron Aqueous Extract Inhibits the Chemically-induced Gastric Cancer Progression in the Wistar Albino Rat

Abstract

Objective(s): Gastric cancer is the first and second leading cause of cancer related death in Iranian men and women, respectively. Gastric cancer management is based on the surgery, radiotherapy and chemotherapy. In the present study, for the first time, the beneficial effect of saffron (Crocus sativus L.) aqueous extract (SAE) on the 1-Methyl -3- nitro -1- nitrosoguanidine (MNNG)-induced gastric cancer in rat was investigated.

Materials and methods: MNNG was used to induce gastric cancer and then, different concentrations of SAE were administered to rats. After sacrificing, the stomach tissue was investigated by both pathologist and flow cytometry, and several biochemical parameters was determined in the plasma (or serum) and stomach of rats.

Results: Pathologic data indicated the induction of cancer at different stages from hyperplasia to adenoma in rats; and the inhibition of cancer progression in the gastric tissue by SAE administration; so that, 20% of cancerous rats treated with higher doses of SAE was completely normal at the end of experiment and there was no rat with adenoma in the SAE treated groups. In addition, the results of the flow cytometry/ propidium iodide staining showed that the apoptosis/proliferation ratio was increased due to the SAE treatment of cancerous rats. Moreover, the significantly increased serum LDH and decreased plasma antioxidant activity due to cancer induction fell backwards after treatment of rats with SAE. But changes in the other parameters (Ca(2+), tyrosine kinase activity and carcino-embryonic antigen) were not significant.

Conclusion: SAE inhibits the progression of gastric cancer in rats, in a dose dependent manner.

Keywords: Anticancer; Crocus sativus; Flow Cytometry; LDH; MNNG; Saffron.

Figure 1

Histopathologic pictures of stomach of all groups of animals in the study. (A) Normal rat stomach. (B) MNNG-administered rat that show hyperplasia: thickening of stomach glandular mucosa and enhancement of cellular population in gastric glands. (C) MNNG-administered rat that show metaplasia: with mucus-producing cells, presence of goblet cells in stomach that is completely abnormal. The mucus producing cells in stomach are hyperplastic too. (D) MNNG-administered rat that show dysplasia: characteristic difference in shape, size, color and dimension of glandular cells plus increase in number and population of them. (E) MNNG-administered rat that show adenoma: with polymorphonuclear cells, increase in cell population, no mitotic figures and no other signs of malignancy like necrosis, hemorrhage, etc. was observed. The cells are well differentiated and nuclei are not that much active, most of them have heterochromatin, but nucleoli are not obvious

Figure 2

Effect of MNNG administration and SAE treatment on the cell cycle status of the stomach tissue of rats that was determined by flow cytometry. (A). Percentage of the cells placed at G0/G1 phase in different groups. (B). Percentage of the cells at S phase. (C). Percentage of the cells at G2/M phase. a = significant difference between group A with labeled groups; b = significant difference between B₁ with labeled group; i = significant difference between B₂ with labeled group, j = significant difference between B₃ with labeled group. (D). Apoptosis Index/ Proliferation Index (AI/PI) ratio. AI/PI ratio in group A was lower than other groups and there was a significant difference (P <0.05) between these groups

Figure 3

Antioxidant capacity of the plasma of all rats at the end of experiment, which was measured by FRAP method. The significant differences between groups are shown as follows: a = significant difference between group A with B1 and B2; i= significant differences between B₄ with B2

Figure 4

LDH activity in the serum of animals before and after treatment with MNNG and/or SAE. There was a significant difference (P<0.05) between group A with other groups. The significant difference (P<0.05) was also exist between group B1 with other groups at the end of experiment. The significant difference (P<0.05) between serum LDH activity of each group before and after SAE treatment was also observed

Figure 5

Evaluation of the total protein content of stomach tissues in different groups of rats. Protein content in groups B1 to B4 was significantly (P<0.05) higher than group A. All represented data are the Mean±SD of the protein content of tissues on mg/g of stomach