来曲唑联合米非司酮对子宫肌瘤模型大鼠的消瘤效果及对TIMPs和PTEN蛋白表达的影响

Antitumor effect of letrozole combined with mifepristone on uterine fibroids model rats and its influence on TIMPs and PTEN protein expression

目的探讨曲唑联合米非司酮对子宫肌瘤模型大鼠的消瘤效果及对基质金属蛋白酶组织抑制因子(TIMPs)和肿瘤抑制蛋白磷酸酶和张力蛋白同源物(PTEN)蛋白表达的影响。方法40只2月龄成年未孕SD大鼠为研究对象,采用雌孕激素双重造模法建立子宫肌瘤模型,选择成模及正常大鼠共36只,按照实验目的分为4组:模型组仅进行生理盐水干预,单一治疗组使用来曲唑进行灌胃治疗,灌胃剂量200 mg·kg^(-1)·d^(-1);联合治疗组同时进行200 mg·kg^(-1)·d^(-1)来曲唑灌胃和2.24 mg·kg^(-1)·d^(-1)米非司酮腹腔内注射,共21 d。对各组大鼠体质量和子宫重量进行称重,并计算子宫系数;采用放射免疫分析法试剂盒检测治疗前后各组大鼠血清中雌二醇和孕激素水平;采用大鼠酶联免疫吸附试验检测治疗前后大鼠血清中TIMPs和PTEN水平;采用蛋白质印迹检测子宫组织TIMPs和PTEN表达。结果(1)与对照组相比,UFs模型组、单一治疗组和联合治疗组大鼠体重显著降低,子宫重量和子宫系数显著增加,差异均有统计学意义(P<0.05);与UFs模型组相比,2个治疗组大鼠体重显著增加,子宫重量和子宫系数显著降低,联合治疗组子宫系数也显著低于单一治疗组,差异均有统计学意义(P<0.05),而两组大鼠体重和子宫重量相比差异无统计学意义(P>0.05)。(2)治疗前,与对照组相比,UFs模型组、单一治疗组和联合治疗组大鼠血清E2、P、TIMPs和PTEN水平均显著降低,差异均有统计学意义(P<0.05)。治疗后,与对照组相比,UFs模型组、单一治疗组和联合治疗组大鼠血清E2、P、TIMPs和PTEN水平均显著降低,差异均有统计学意义(P<0.05);与UFs模型组相比,2个治疗组大鼠血清E2、P、TIMPs和PTEN水平均显著升高,联合治疗组血清E2、P、TIMPs和PTEN水平也显著高于单一治疗组,差异均有统计学意义(P<0.05)。(3)与对照组相比,UFs模型组、单一治疗组和联合治疗组大鼠子宫组织TIMPs和PTEN表达均显著降低,差异均有统计学意义(P<0.05);与UFs模型组相比,2个治疗组子宫组织TIMPs和PTEN表达均显著升高,联合治疗组子宫组织TIMPs和PTEN表达也显著高于单一治疗组,差异均有统计学意义(P<0.05)。结论来曲唑联合米非司酮应用于子宫肌瘤大鼠具有显著治疗效果,可显著减小瘤重,促进大鼠体重恢复,并改善血清雌孕激素水平,其作用机制可能与激活抑癌相关蛋白TIMPs和PTEN表达有关。

Objective To explore the anti-tumor effect of trazole combined with mifepristone on uterine fibroids model rats and its influence on the expression of tissue inhibitor of matrix metalloproteinases(TIMPs)and phosphatase and tensin homolog(PTEN)protein.Methods Forty 2-month-old adult non-pregnant SD rats were used as the research objects.The uterine fibroids model was established by the double modeling method of estrogen and progesterone.A total of 36 model and normal rats were selected and divided into 4 groups according to the experimental purpose:the model group was treated with physiological saline only,and the single treatment group was treated with letrozole for intragastric treatment,with an intragastric dose of 200 mg·kg^(-1)·d^(-1);The combined treatment group received 200 mg·kg^(-1)·d^(-1) letrozole and 2.24 mg·kg^(-1)·d^(-1) mifepristone intraperitoneally for 21 days.The body weight and uterine weight of the rats in each group were weighed,and the uterine coefficient was calculated.The levels of estradiol and progesterone in serum of rats in each group before and after treatment were detected by radioimmunoassay kit.The levels of TIMPs and PTEN in rat serum before and after treatment were detected by rat enzyme-linked immunosorbent assay.The expression of TIMPs and PTEN in uterine tissue were detected by Western Blotting.Results(1)Compared with the control group,the weight of the rats in the UFs model group,the single treatment group and the combination treatment group was significantly reduced,and the uterine weight and uterine coefficient were significantly increased,the differences were statistically significant(P<0.05);Compared with the UFs model group,the weight of rats in the two treatment groups increased significantly,and the uterine weight and uterine coefficient were significantly reduced,the differences were statistically significant(P<0.05).The uterine coefficient of the combined treatment group was also significantly lower than that of the single treatment group,the differences were statistically significant(P<0.05);There was no significant difference in body weight and uterine weight between the two groups(P>0.05).(2)Before treatment,compared with the control group,the levels of serum E2,P,TIMPs and PTEN of rats in the UFs model group,the single treatment group and the combination treatment group were significantly lower,the differences were statistically significant(P<0.05).After treatment,compared with the control group,the levels of serum E2,P,TIMPs and PTEN of rats in the UFs model group,the single treatment group and the combination treatment group were significantly reduced,the differences were statistically significant(P<0.05);Compared with the UFs model group,the levels of serum E2,P,TIMPs and PTEN of rats in the two treatment groups were significantly increased,and the levels of serum E2,P,TIMPs and PTEN of the combined treatment group were also significantly higher than those of the single treatment group,the differences were statistically significant(P<0.05).(3)Compared with the control group,the expressions of TIMPs and PTEN in the uterus tissues of the UFs model group,single treatment group and combination treatment group were significantly reduced,the differences were statistically significant(P<0.05);Compared with the UFs model group,the expressions of TIMPs and PTEN in uterine tissues of the two treatment groups were significantly increased,the differences were statistically significant(P<0.05),and the expressions of TIMPs and PTEN in uterine tissues of the combined treatment group were also significantly higher than those of the single treatment group,the differences were statistically significant(P<0.05).Conclusion The combination of letrozole and mifepristone in rats with uterine fibroids has a significant therapeutic effect,which can significantly reduce tumor weight,promote weight recovery in rats,and improve serum estrogen and progesterone levels.Its mechanism of action may be related to the activation of the expression of tumor suppressor-related proteins TIMPs and PTEN.