黑升麻的雌激素活性及其对人类乳腺癌MCF-7细胞雌激素受体水平的影响

为了检测黑升麻 (CR)的雌激素活性及其对人类乳腺癌 MCF- 7细胞雌激素受体 (ERs)水平的影响 ,未成年雌性小鼠按体重以 75、15 0和 30 0 m g/ kg的 CR灌胃 14d,观察处于动情期的时间 ,第 15天处死后取子宫和卵巢称重。以 MTT实验筛选 CR的最佳细胞作用剂量 ,观察对照组、CR组 (4.75μg/ L )和 17β-雌二醇组 (0 .3nmol/ L)的细胞生长曲线 ,并在流式细胞仪上用间接免疫荧光法测定各组 ERs水平。结果发现子宫重量随 CR剂量增加而增加 ,30 0 mg/ kg的 CR可显著增加动情天数 (P<0 .0 5 )。 4.75 μg/ L CR对 MCF- 7细胞的促增殖作用最强 ,达 6 4.7% ;CR组和 17β-雌二醇组的细胞群体倍增时间 (TD)分别为 32 .1h和31.7h,明显低于对照组 (TD=35 .3h) ;4.75 μg/ L 的 CR可显著增加细胞的 ERs水平 (P<0 .0 1)。结果表明CR具有雌激素活性 ,其升高 ERs水平的作用可能是该物质可治疗更年期综合征的机制之一。

Estrogenicity of black cohosh (cimicifuga racemosa) and its effect on estrogen receptor level in human breast cancer MCF-7 cells

The estrogenicity of Black Cohosh (Cimicifuga racemosa, CR) was tested in vivo and in vitro and its effect on estrogen receptor (ER) level of human breast cancer MCF-7 cells were investigated. Based on the body weight of animals, 75, 150 and 300 mg/kg of CR were administered by tube feeding to immature female mice for 14 days. Estrus was observed and the uterine and ovary weights of mice were measured. The optimal dose of CR for the growth of MCF-7 cells was screened by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay. Subsequently, Growth curves of MCF-7 cells in blank control, 4.75 micrograms/L of CR and 0.3 nmol/L of 17 beta-estradiol groups were observed for 5 days. ER level in MCF-7 cells was analyzed by indirect immunofluorescence assay in flow cytometry. The results showed that uterine weight increased with the increasing dosage of CR and the days of estrus was significantly prolonged in the 300 mg/kg group (P < 0.05). The concentration of CR at 4.75 micrograms/L showed the strongest enhancement effects (64.7%). The doubling time (TD) of cell growth in CR group and 17 beta-estradiol group were 32.1 h and 31.7 h respectively, which were shorter than that of blank control (TD = 35.3 h). Additionally, 4.75 micrograms/L of CR significantly increased ER levels compared with the blank control (P < 0.01). Taking all the results together, CR has an estrogen-like action. The enhancing effect of CR on ER level is one of the potential mechanisms involved with its therapeutic role in climacteric syndrome.