A reporter gene assay for the detection of phytoestrogens in traditional Chinese medicine.
Bupleurum & Peony Formula (Jia Wei Xiao Yao San) is a herbal formula which possesses a clinical history for the treatment of menopausal syndrome and menstrual irregularity. The present investigation reports the ability to monitor the formula's phytoestrogen content that will allow for the implementation of a standardization protocol that is based on a quantifiable biological response. Utilizing an oestrogen-sensitive chimeric receptor/reporter gene element which has been stably transfected into HeLa cells, the botanical formula was shown to induce the expression of the reporter gene, luciferase, in a dose dependent manner. Pretreatment of the HeLa cells with the botanical formula produced a 5-fold increase in bioluminescence compared with the control. Additionally, our studies showed that the response of the cells, when challenged by the botanical formula, was oestrogen specific. Pretreatment of the cells with tamoxifen effectively blocked the activation of the chimeric oestrogen receptor by the botanical formula. The cell line provides a sensitive assay that can easily detect the presence of phytoestrogens in complex botanical formulas.
Miller-Martini
D M
DM
The Biotechnology Research Institute, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China.
Chan
R Y
RY
Ip
N Y
NY
Sheu
S J
SJ
Wong
Y H
YH
D000276:Adjuvants, Immunologic; D029972:Bupleurum / Q000737:chemistry*; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D004968:Estrogens, Non-Steroidal / Q000302:isolation & purification*; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D007529:Isoflavones; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements; D008516:Medicine, Chinese Traditional; D008593:Menopause / Q000187:drug effects*; D048789:Phytoestrogens; D008517:Phytotherapy; D010936:Plant Extracts / Q000737:chemistry; D028321:Plant Preparations; D011960:Receptors, Estrogen / Q000235:genetics* / Q000378:metabolism; D012680:Sensitivity and Specificity; D013629:Tamoxifen / Q000494:pharmacology; D014162:Transfection
D000276:Adjuvants, Immunologic; D004365:Drugs, Chinese Herbal; D004968:Estrogens, Non-Steroidal; D007529:Isoflavones; D048789:Phytoestrogens; D010936:Plant Extracts; D028321:Plant Preparations; D011960:Receptors, Estrogen; D013629:Tamoxifen; D004958:Estradiol; D008156:Luciferases