Experimental Study Of Inhibitory Effect Of Growth On Rats Bladder Tumor And Apoptosis-Induced By Allicin

Objective: explore the treatment on bladder cancer of Allicin, reveal the relationship with apoptosis, further find the mechanism of its anti-tumor, provide a new method for the treatment of bladder tumor.Methods:1. The establishment of rat modes: rat bladder tumor models were established by perfusing N-methyl-N-nitrosourea into bladder.2. 60 tumor-bearing rats were randomly divided into four groups: saline group, cisplatin group, low concentration of allicin group, high concentration of allicin group; every group was composed of 15 rats. Route of administration: intraperitoneal injection.one time every other day, seven times in all. During the raised periods, observed the life of rats and weight changes, all rats were sacrificed after injection of four times, examined the changes of bladder total weights,HE staining was apllied to bladder tumors of four groups.3.Apoptosis were detected in bladder tumor tissues by TUNEL method, RT-PCR and immunohistochemistry methods were apllied to detect caspase-3 mRNA and protein expression in the tumor tissues, revealed the apoptosis mechanism of allicin in bladder tumor.Results:1. Histopathological changes of MUN induced rats bladder tumors:l2 weeks after MNU perfusion, five rats were selected randomly,and killed, we found that bladder walls were thickening, pale, new cauliflower-shaped formation, the overwhelming majority of the tumor for more serious cases, reached the level of full bladder. HE staining showed transitional cell carcinoma with myometrial invasion. No other organs of the groups were abnormal. Accordingly, we selected tumor-bearing rats as the following experimental animals.2. Anti-tumor effect of allicin:After administration, there were no differences of rats weight in the normal saline group, low concentrations of allicin group (p>0.05), but rats weight increased with the age in high concentrations of allicin group ,compared to administration before, the differences were statistically significant (P <0.05), and rats had good appetites, normal activities, glossy fur in the above-mentioned groups. But rats weight of cisplatin group decreased with the growth of age, contrasted to other groups, the differences were significant (P<0.05), and rats had shown significant anorexia, slow activities, abusive dark fur. Rats bladder total weight of saline group were higher than other groups, it was caused by MNU intravesical bladder cancer to increase the weight of the bladder; low concentration and high concentration of allicin group, differences were significant (p<0.05); there were no significant differences between cisplatin group and high concentration of allicin group (p>0.05). Before and after treatment, there were little histopathological changes in bladder tumor tissue of saline group , and tumor cells growed brisk, no death; and different degrees bladder tumor necrosis occurred in cisplatin group and different concentrations of allicin group. Apoptosis were difficult to monitor in tumor cells by light microscopy . 3. Apoptosis and caspase-3 mRNA and protein expression in bladder tumors of four groups:All the groups were detected in the nuclei of tumor cells in varying degrees with brown staining by situ TUNEL method. Compared to saline group, the AI of other groups were significantly higher (P<0.05). And compared to cisplatin group,low concentration of allicin group, there was more obvious role in apoptosis of high concentration allicin group , the differences were statistically significant (P<0.05) .We found that allicin could upregulate the level of casapse-3 mRNA in rats bladder tumors.We used Immunohistochemical SP method to detect caspase-3 protein expression in rat bladder tumor tissues, we observed that the caspase-3 positive products were brown in the membrane and cytoplasm by optical microscopy. we also found that allicin could upregulate the level of casapse-3 protein in bladder tumors.Conclusion:1. MNU intravesical induced bladder tumors in rats, the method is reliable, it is a good way to establish bladder tumor model.2. Allicin is not only reliable security, but also have significant inhibitory effect on bladder tumors of rats.3. Inducing tumor cells apoptosis is one of the ways for anti-tumor of allicin, and increasing expression levels of caspase-3 mRNA and protein may be one of ways for tumor cells apoptosis.