基于caspase 3、caspase 9表达探讨桂枝茯苓丸治疗小鼠子宫腺肌病的药效评价

Evaluation of Efficacy of Guizhi Fuling Pill in Treatment of Adenomyosis in Mice Based on Expressions of Caspase 3 and Caspase 9

目的观察经不同处理的桂枝茯苓丸治疗小鼠子宫腺肌病疗效及其子宫内膜细胞凋亡情况,评价桂枝茯苓丸有效成分提取方式及药效表达,为将来开展桂枝茯苓丸治疗子宫腺肌病的药效物质发现和辨识奠定基础。方法选用他莫西芬药物诱导法制备ICR小鼠子宫腺肌病动物模型,随机分为空白组、模型组、桂枝茯苓丸醇提物组、桂枝茯苓丸超微粉组、中成药组和米非司酮组,干预15 d后取材,通过各组子宫一般形态观察、HE病理评分、caspase 3、caspase 9免疫组化蛋白阳性表达情况判断疗效。结果各治疗组HE病理评分皆低于模型组,caspase 3、caspase 9蛋白阳性表达高于模型组,皆有统计学意义,但各治疗组间差异无明显统计学意义。结论桂枝茯苓丸醇提物和超微粉可与中成药及米非司酮一样发挥疗效,通过促进子宫内膜细胞凋亡,抑制异位内膜侵袭性而缓解子宫腺肌病的发展,醇提和超微粉碎技术可作为有效提取方式最大程度保留桂枝茯苓丸中有效成分。

Objective To observe the therapeutic effect of different treatments of Guizhi Fuling Pills on adenomyosis and apoptosis of endometrial cells in mice and evaluate the extraction method of active ingredients of Guizhi Fuling Pills for the treatment of adenomyosis in the future and provide the foundation for the discovery and identification of pharmacodynamic substances.Methods The animal model of adenoma of ICR was prepared by tamoxifen drug induction method,and the micewere randomly divided into blank group,model group,Guizhi Fuling Pill extract group,Guizhi Fuling Pill ultrafine powder group,Chinese patent medicine group and mifepristone group.The drug was obtained after 15 days of intervention,and the positive effects of uterine morphological observation,HE pathological score,immunohistochemical protein expressions of caspase 3 and caspase 9 were evaluated.Results The pathological scores of HE in each treatment group were lower than those in the model group.The positive protein expressions of caspase 3 and caspase 9 was higher than that in the model group,and there was statistical significance,but there was no statistical difference among the treatment groups.Conclusion The alcohol extract and ultrafine powder of Guizhi Fuling Pill can exert the same effect as Chinese patent medicine and mifepristone,and alleviate the development of adenomyosis by promoting the apoptosis of endometrial cells and inhibiting the invasiveness of ectopic endometrium.And ultra-fine pulverization technology can be used as an effective extraction method to retain the active ingredients in Guizhi Fuling Pill.