四物汤及组方中药植物雌激素活性的实验研究

Study on the Phytoestrogenic Effects of SiWu Decoction and its Ingredient Herbs

目的:首先考查四物汤及其组方中药对性未成熟雌性SD大鼠子宫系数的影响,然后考查含药血清对雌激素受体亚型ERα阳性人乳腺癌MCα衡和随机的原则分为7组(正常对照组、己烯雌酚组、四物汤组、熟地组、白芍组、当归组和川芎组),分别每天分早、晚两次灌胃给药,给药均持续4天,第5天腹主动脉取血,分离血清,-20℃贮存备用,断颈处死动物后,分离子宫并称重,计算子宫系数;应用MTT法检测药物血清对MCF-7细胞的增值率、流式细胞技术检测细胞周期和ERα蛋白表达。结果:熟地、白芍和当归组的幼鼠子宫系数均高于正常对照组(P<0.05);川芎组和四物汤组的幼鼠子宫系数与正常对照组相比,差异无统计学意义(P>0.05);白芍、当归和川芎等中药的药物血清均可显著促进MCF-7细胞体外增殖(P<0.01),熟地可促进MCF-7细胞体外增殖(P<0.05);熟地、当归和川芎等中药的药物血清均可显著促使MCF-7细胞周期S期细胞数增加(P<0.01),白芍的药物血清可促使MCF-7细胞周期S期细胞数增加(P<0.05);而在本实验条件下四物汤复方的含药血清使MCF-7细胞数略有增加,但无统计学意义(P>0.05);当归和川芎可提高ERα蛋白的表达水平(P<0.05),熟地、白芍和四物汤也可提高ERα蛋白的表达水平,但与正常对照组相比,差异无统计学意义(P>0.05)。结论:熟地、白芍、当归等中药有植物雌激素活性,本实验配伍比例条件下,四物汤复方的植物雌激素活性弱于其组方的各味中药。

Objective：To investigate the phytoestrogenic- like effects of SiWu Decoction and its ingredient herbs. Method：Immature female SD rats weighting （70 ±5 ）g were randomly divided into seven groups：normal control group, positive control group and five Chinese medicine groups. The rats in different groups were treated for four days. On the fifth day, animals were sacrificed and uteri were separated and weighed. The blood was collected, and serum was separated. The effects of the pharmacological serum on proliferation assay with human breast cancer cell line （MCF- 7 ）were observed by MTr assay. Cell-cycle analyses were performed with propidium iodide staining by flow eytometry. The expressions of subtypes-estrogen receptor α（ERα）were detected by flow cytometry. Results：Radix rehmanniae preparata, Radix paeoniae alba and Radix angelieae sinensis could increase the immature rats＇ uterus wet weight and the ratio of uterus to body weight （ P 〈 0. 05 ）. SiWu Decoction and Rhizoma ehuanxiong could not increase the immature rats＇ uterus wet weight and the ratio of uterus to body weight （ P 〉 0.05 ）. The pharmacological serum of Radix paeoniae alba, Radix angelieae sinensis and Rhizoma ehuanxiong stimulated proliferation of MCF - 7 cell respectively compared with normal control group（ P 〈 0.01 ）. The pharmacological serum of Radix rehmanniae preparata stimulated proliferation of MCF - 7 cell compared with normal control group（ P 〈 0.05 ）. The amount of cell in Phase S and proliferation index increased with the serum of Radix rehmauniae preparata,Radix angelieae sinensis and Rhizoma chuanxiong（ P 〈 0. 01 ）. Proliferation index increased with the serum of Radix paeoniae alba（ P 〈 0.05 ）. The pharmacological serum of SiWu Decoction could, to a certain extent, increase the amount of MCF- 7 cells when its ingredients were at the specific ratio. However, no statistical significance was detected（ P 〉 0. 05 ）. The pharmacological serum of Radix angelicae sinensis and Rhizoma ehuanxiong could increase the expressions of subtypes-estrogen receptor α （ERα） （ P 〈 0. 05 ）. Conclusion ： Radix rehmanniae preparata, Radix paeoniae alba and Radix angelicae sinensis possess phytoestrogenic effects. SiWu Decoction possesses less phytoestrogenie effects than each of its ingredients when the ingredients of the decoction were at the specific ratio.