Division of Life Science, Center for Chinese Medicine and State Key Laboratory of Molecular Neuroscience, The Hong Kong University of Science and Technology, Clear Water Bay Road, Hong Kong, China.
J Ethnopharmacol. 2012 Aug 30;143(1):109-15. doi: 10.1016/j.jep.2012.06.009. Epub 2012 Jun 16.
Er Zhi Wan (EZW), a Chinese medicinal preparation, has been used clinically for treating menopausal syndrome for its kidney-invigorating function, which contains simply two herbs, Ecliptae Herba (EH) and Ligustri Lucidi Fructus (LLF). Although this herbal extract has been used for many years, there is no scientific basis about its effectiveness on menopausal symptom. Here, we aimed to evaluate the estrogenic activities of EZW and to study the compatibilities of two herbs including different processed-LLF in single and mixed preparation of EZW. Moreover, the weight ratio of EH to LLF in EZW was determined according to their estrogenic activities.
The extractions of LLF, processed-LLF and EH were prepared separately by extracting the powders with water, 50% alcohol or 95% alcohol. Steamed-LLF and EH were extracted separately, or together, in preparing EZW extracts. A promoter-reporter construct (pERE-Luc) containing three repeats of estrogen responsive elements (ERE) was stably transfected into MCF-7 cells, and this stable breast cancer cell line was used to determine the estrogenic property. The cell proliferation was measured by MTT assay.
The results showed that EZW could significantly induce the expression of luciferase driven by an estrogen responsive element in a pERE-Luc vector. The proliferation of MCF-7 cells was not altered by this herbal treatment. The best preparation of EZW was from: (i) LLF was firstly steamed over water and then dried to make steamed-LLF; and (ii) steamed-LLF and EH were extracted separately by 95% alcohol and then mixed together according to a weight ratio of 1:1.
Under the optimized extracting method, EZW possessed robust effect in activating the estrogenic activity, but which did not alter the proliferation of cultured MCF-7 cells. Thus, EZW is an effective and safe estrogenic herbal extract.
二至丸（EZW）是一种中药制剂，因其补肾功能，已在临床上用于治疗更年期综合征。它仅包含两种草药，旱莲草（EH）和女贞子（LLF）。虽然这种草药提取物已经使用了很多年，但它对更年期症状的有效性没有科学依据。在这里，我们旨在评估 EZW 的雌激素活性，并研究两种草药包括不同炮制的女贞子在 EZW 的单药和混合制剂中的相容性。此外，根据其雌激素活性，确定 EZW 中 EH 与 LLF 的重量比。
分别用水、50%酒精或 95%酒精提取 LLF、炮制的 LLF 和 EH 的粉末。将蒸制的 LLF 和 EH 分别提取，或一起提取，制备 EZW 提取物。含有三个雌激素反应元件（ERE）重复的启动子-报告基因构建体（pERE-Luc）稳定转染 MCF-7 细胞，该稳定的乳腺癌细胞系用于确定雌激素性质。通过 MTT 测定法测量细胞增殖。
结果表明，EZW 可显著诱导含有雌激素反应元件的 pERE-Luc 载体中荧光素酶的表达。这种草药处理不会改变 MCF-7 细胞的增殖。EZW 的最佳制备方法是：（i）首先将 LLF 用水蒸制，然后干燥制成蒸制的 LLF；（ii）蒸制的 LLF 和 EH 分别用 95%酒精提取，然后按照 1:1 的重量比混合在一起。
在优化的提取方法下，EZW 具有激活雌激素活性的强大作用，但不会改变培养的 MCF-7 细胞的增殖。因此，EZW 是一种有效且安全的雌激素草药提取物。
