| Purpose: To study the effects of SIN on tumor immunity and apoptosis of MNU cancer in male SD rats induced by N-methyl nitrosa(N-methyl-nitrosourea,MNU)by bladder perfusion of Sinomenine and preliminary discuss the anti-tumor effect of SIN in bladder cancer transition.Materials and methods: In this study,male SD rats were used as experimental subject and male SD rats were divided into 5 groups on average,with 10 in each group,using randomized contrast principle.The groups were divided into contrast principle(Group A),model group(Group B),Sinomenine group(Group C),Sinomenine and half-volume BCGG group(Group D),BCG group(Group E)and other 5 groups.In addition to Group A,the bladder perfusion was carried out for male SD rats with MNU solution the remaining groups to establish a model of bladder cancer in rats.After 10 weeks of modeling,the bladder perfusion was carried out,0.2 ml of physiological saline was perfusion in both A and B groups and 0.2 ml of aqueous solution was perfusion in group C.On the basis of group C,1.25mg/ml bcg0.2ml was added to group D and 2.5mg/ml bcg0.2ml was added to group E.The above operations are carried out once a week for 4 weeks.Rats were executed immediately after extraction of vein blood and bladder tissue was used immuno-printing method to detect each group of cytotoxic T lymphocyte-related antigen4(CTLA-4),programmed cytokine die factor 1(programmed cell body-1,PD-1)and caspace-3.The ELISA method detected the level of serum soluble co-stimulator B7-H3(co-stimulus B7-H3,B7-H3)and the HE dying method assessed the bladder pathology results of rats.Results: 1,the expression level of CTLA-4 in each group was 12.30±2.65,33.54±4.92,26.04±4.57,23.85±4.71 and 17.97±4.48;where,CTLA-4 in Group A has the lowest expression level and that in Group B has the highest expression;The expression level of CTLA-4 in Group A was significantly lower than that in the remaining 4 groups(P less than 0.05);The expression level of CTLA-4 in Group B was significantly higher than that in the remaining 4 groups(P less than 0.05);the expressed level of CTLA-4 in Group E was significantly higher than group C(P 0.05).The expression level of PD-1 in each group was 15.53 ± 4.72,36.54 ± 6.45,28.30 ± 5.18,21.59 ± 7.04 and 19.51 ± 5.74,respectively.The expression of PD-1 in group A was the lowest and that in Group B was the highest(P < 0.05).The expression level of PD-1 in Group B was significantly higher than that in the other four groups(P < 0.05);the expression level of PD-1 in Group C was significantly lower than that in Group D and E(P < 0.05).The expression level of B7-H3 in each group was 12.30 ± 2.65,33.54 ± 4.92,26.04 ± 4.57,23.85 ± 4.71 and 17.97 ± 4.48,respectively.Among them,the expression level of B7-H3 was the lowest in Group A and the highest in Group B;the expression level of B7-H3 in Group A was significantly lower than that in the other four groups(P < 0.05);the expression level of B7-H3 in Group B and C was significantly higher than that in Group D and E(P < 0.05).2.The expression of Caspase-3 was 0.84 ± 0.26,1.83 ± 0.51,1.28 ± 0.35,1.21 ± 0.32 and 1.08 ± 0.37 in each group,among which the expression level of Caspase-3 was the lowest in Group A and the highest in Group B;the expression of Caspase-3 in Group A was significantly lower than that in the other four groups(P < 0.05);the expression of Caspase-3 in Group B was significantly higher than that in the other four groups(P < 0.05);the expression of Caspase-3 in Group C was significantly higher than that in Group D and Group E(P < 0.05)05).3.The results of Group A were normal bladder tissue and that in Group B,C,D and E were diagnosed as bladder cancer.The average level of pathological examination in each group were 0.79 ± 0.31,3.18 ± 1.33,2.31 ± 0.84,1.39 ± 0.65 and 1.16 ± 0.52,respectively.Among the five groups,the HE score of bladder inflammation and disease examination in Group A was the lowest and that in Group B was the highest;the HE score of bladder inflammation and disease examination in Group A was significantly lower than that in the other four groups(P < 0.05);the HE score of bladder inflammation and disease examination in Group B was significantly higher than that in the other four groups(P < 0.05);the HE score of bladder inflammation and disease examination in Group C was significantly higher than that in Group D and Group E respectively(P < 0.05).Conclusion: 1.The SIN can significantly the expression of inhibit MNU-induced bladder cancer CTLA-4 and PD-1 and may inhibit bladder cancer by lowering the level of immune checkpoint molecular expression of bladder cancer;the SIN can significantly inhibit the expression of MNU-induced bladder cancer caspase-3,accelerate the apoptosis process of bladder cancer cells,which may also be a mechanism for SIN to inhibit the progression of bladder cancer;... |
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