Aim To study the phytoestrogenic effect and its mechanism of isopsoralen in estrogen receptor (ER) positive T47D and ER negative MDA-MB231 breast cancer cell line.Methods The proliferation rate and cell cycle distribution of T47D and MDA-MB231 cells influenced by isopsoralen were analyzed by MTT and flow cytometry assay respectively.PS2 mRNA level in T47D cells was quantified by Real-time RT-PCR assay.PS2 expression was measured by flow cytometry.Estrogen receptor antagonist ICI180,782 was employed as a tool.Results The proliferation rates and indexes of T47D cells treated with 1?10-6 mol?L-1 and 1?10-7 mol?L-1 isopsoralen were increased.These effects could be blocked by ICI182,780.Meanwhile,isopsoralen didn't show stimulative effects on proliferation rate of MDA-MB231 cells.Real-time RT-PCR and flow cytometry results showed that isopsoralen at 1?10-6 mol?L-1 and 1?10-7 mol?L-1 could induce elevation of pS2 mRNA and protein expression level in T47D cells.These effects could be partly blocked by ICI182,780.Conclusion Isopsoralen has phytoestrogenic effect and its effect is attained mainly via ER pathway.

This study was aimed to observe the influence of β-sitosterol (BSS) on estrogen receptor (ER) positive the human breast cancer cell line T47D and to study its mechanisms. ER antagonist ICI182 780 was employed to observe the influence on the proliferation. Proliferations of T47D cells influenced by different concentrations of BSS were analyzed by MTT assay. Cell cycle analyses were examined by flow cytometry. The protein expression of cyclin D1 was measured by western blot analysis and cyclin D1 mRNA was quantified by real-time PCR assay. The results showed that BSS in high dose exhibited significant inhibitory effects that were partly antagonized by ICI182 780 and decreased the proliferative index on T47D cells. However, BSS in low dose obviously promoted the proliferation that was completely inhibited by ICI182 780 and increased the proliferative index on T47D cells. The mRNA and protein levels of cyclin D1 were increased in low-dose BSS. The effect was blocked by ICI182 780. It was concluded that BSS in low concentration had phytoestrogenic effect by up-regulating the expression of cyclin D1 via ER pathway.

OBJECTIVEResearch on the phytoestrogenic effect and its possible mechanism of formononetin.

METHODTo evaluate the estrogenic effect and mechanisms of formononetin through the test of its influence on proliferation and ER subtype expression of T47D cells.

RESULTThe proliferation rates of T47D cells treated with 1 x 10(-7) -1 x 10(-6) mol x L(-1) formononetin were not increased. On the influence of ICI182, 780, the proliferation rates of T47D cells treated with 1 x 10(-7) 1 x 10(-6) mol x L(-1) formononetin were decreased. Formonenetin could induce the augment of ERalpha expression significantly of T47D.

CONCLUSIONFormonenetin has phytoestrogenic effect Formonenetin can not accelerate ER(+) T47D cell proliferation. But the expression level of ERalpha subtype in T47D cells change significantly with certain concentrations of formonenetin.

Breast Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Humans ; Isoflavones ; pharmacology ; Phytoestrogens ; pharmacology ; Receptors, Estrogen ; genetics ; metabolism

OBJECTIVETo study the phytoestrogenic effects and their possible mechanisms of Jiaoai tang and Shenqi Jiaoai tang through the tests in mice and ER (+) MCF7 cells.

METHODSixty kunming mice weighing 9-12 g were randomly divided into 6 groups: solvent control group (administrated equal dose of diswater), diethylstilbestrol control group (administrated diethylstilbestrol at a dose of 0.35 mg x kg(-1) x d(-1) and 4 Chinese medicine treated groups (administrated low or high doses of Jiaoai tang and Shenqi Jiaoai tang at 2.5 g x kg(-1) x d(-1) or 5.0 g x kg(-1) x d(-1) respectively). After administration for 4 days, the mice were sacrificed; uterus was removed and weighed, uterus rate was calculated. The blood serum was also separated. The proliferation rate of MCF7 cells influenced by Jiaoai tang and Shenqi Jiaoai tang was determined by MIT assay. PS2, ERalpha and ERbeta mRNA expression was quantified by Real-time PCR assay. Estrogen receptor antagonist ICI182, 780 was employed as a tool.

RESULTAdministration of Jiaoai tang and Shenqi Jiaoai tang at high dose significantly increased uterus rate in mice (P < 0. 05). The pharmacological serum from two high-dosage groups of Chinese herbal medicine decoction significantly enhanced proliferation rate of MCF7 cells (P < 0.05 or 0.01), while their effects were blocked by ICI182, 780 (P < 0.05 or 0.01). The pharmacological serum could cause elevation of pS2 level (P < 0.01) which would be obviously inhibited by ICI182, 780 (P < 0.01). ERalpha and ERbeta mRNA levels were also elevated significantly (P < 0.05 and P < 0.01 respectively).

CONCLUSIONJiaoai tang and Shenqi Jiaoai tang have phytoestrogenic effects, which were attained via ER pathway. They can also increase the mRNA levels of estrogen receptor subtypes, especially ERbeta.

Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; Estrogen Receptor alpha ; genetics ; metabolism ; Estrogen Receptor beta ; genetics ; metabolism ; Female ; Gene Expression ; drug effects ; Mice ; Phytoestrogens ; pharmacology ; Random Allocation ; Signal Transduction ; drug effects ; Uterus ; drug effects ; metabolism

OBJECTIVETo evaluate the protective effects of siwuheji on cisplatin-induced ovarian impairing and explore its possible mechanisms in mice.

METHODThe mice received cisplatin ip (7 mg kg(-1)) for 7 days, while fed with siwuheji for 14 days. The changes of estrous cycle, ovary and uterus coefficient, levels of estrogen in serum and morphology of ovary were recorded.

RESULTFrom the third day of cisplatin injection, the estrous cycle of the model group stagnated in interphase. On the thirteenth day, part of the mice fed with siquheji started with change of estrous cycle. The ovary coefficient of the model group was less than the control group, the experimental groups were no differences compared with the control group and had significant differences with the model group. The levels of E2 and P of model group were lower than the control group, but no statistical significance. The level of E2 with high-dose Siwuheji group was higher than model group, and the level of P of the low-dose group was higher than model group, but both no statistical significances. The FSH/LH ratio of model group was significantly higher than the control group, low-dose group and medium-dose group of siwuheji resumed to normal levels, and had a significantly change compared with the model group. Ovarian biopsy of model group showed the decrease in the number of developing follicle at all levels, and increase in the number of Atresia of ovarian. There were more growing follicte in the large, medium-dose group.

CONCLUSIONCisplatin can induce mice ovarian injury and functional decline, on the other hand, siwuheji can improve cisplatin-induced ovarian dysfunction.

Animals ; Cisplatin ; adverse effects ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Mice ; Mice, Inbred ICR ; Ovarian Diseases ; chemically induced ; drug therapy ; physiopathology ; Ovary ; drug effects ; physiopathology ; Random Allocation ; Uterus ; drug effects ; physiopathology

OBJECTIVETo explore the phytoestrogenic-like effects of four kinds of Chinese medicine including Radix rehmanniae preparata, Radix Paeoniae Alba, Radix Angelicae Sinensis, Rhizoma Chuanxiong.

METHODSixty immature female SD rats weighting (70 +/- 5) g were randomly divided into six groups: normal control group, positive control group and 4 Chinese medicine groups. The rats in different groups were treated for 4 days. On the fifth day, animals were sacrificed and uteri were separated solely and weighed. The blood was collected, and serum was separated. The effect of the pharmacological serum on proliferation assay with human breast cancer cell line (MCF7) by MTT method. Cell-cycle analyses were carried out with propidium iodide staining by flow cytometer. The expressions of subtypes-estrogen receptor-alpha (ERalpha) were detected by flow cytometry.

RESULTRadix Rehmanniae Preparata, Radix Paeoniae Alba and Radix Angelicae Sinensis could increase the immature rat's uterus wet weight and the ratio of uterus to body weight (P < 0.05). The pharmacological serum of the four kinds of Chinese medicine stimulated proliferation of MCF7 cell respectively compared with normal control group (P < 0.01). The cell cycle was impulsed from G1 to S, DNA synthesizing was inhanced, and PI was also increased. The pharmacological serum of Radix Angelicae Sinensis and Rhizoma Chuanxiong could increase the expressions of (ERalpha) (P < 0.05).

CONCLUSIONRadix Rehmanniae Preparata, Radix Paeoniae Alba and Radix Angelicae sinensis have phytoestrogenic effects. But the data werent consistent with in vitro and in vivo assay of Rhizoma Chuanxiong.

Animals ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Estrogen Receptor alpha ; genetics ; metabolism ; Female ; Phytoestrogens ; administration & dosage ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Uterus ; cytology ; drug effects ; metabolism

OBJECTIVETo investigate phytoestrogenic effects of ferulic acid in ER-positive T47D and ER-negative MDA-MB231 cells in culture.

METHODT47D and MDA-MB231 human breast cancer cells were treated with ferulic acid and examined cell proliferation by means of MTT assay. Cell cycle distribution, ERalpha and ERbeta expression were treated by flow cytometer. The pS2 mRNA expressions were detected by real-time fluorescence quantitative PCR.

RESULTThe proliferations were enhanced significantly by treatment with ferulic acid on T47D cells and the proliferation effects were inhibited by adding Faslodex (1 x 10(-8) mol x L(-1)). However, there was no significant difference on the proliferation in MDA-MB-231 cells compared with solvent control group by both treatment with ferulic acid and co-treatment with Faslodex (1 x 10(-8) mol x L(-1)). Ferulic acid stimulated the amount of T47D cells in phase S and proliferation index increased significantly. The effects were inhibited by treatment with Faslodex (1 x 10(-8) mol x L(-1)), and the amount of cells in phase S and proliferation index decreased, the amount of cells in G0/G1 phase increased, cell cycle of T47D was arrested in G0/G1 phase. Ferulic acid up-regulated pS2 mRNA expressions and increased the level of ERalpha protein expression in T47D cells. Ferulic acid did not show remarkable effect to the level of ERbeta protein expression in T47D cells.

CONCLUSIONFerulic acid possessed phytoestrogenic effect by up-regulating pS2 gene expression and the receptor subtype of ERalpha.

Breast Neoplasms ; chemistry ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Coumaric Acids ; pharmacology ; Estrogen Receptor alpha ; analysis ; Estrogen Receptor beta ; analysis ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; RNA, Messenger ; analysis ; Trefoil Factor-1 ; Tumor Suppressor Proteins ; genetics

The rapid and accurate authentication of traditional Chinese medicines(TCMs)has always been a key scientific and technical problem in the field of pharmaceutical analysis.Herein,a novel heating online extraction electrospray ionization mass spectrometry(H-oEESI-MS)was developed for the rapid and direct analysis of extremely complex substances without the requirement for any sample pretreatment or pre-separation steps.The overall molecular profile and fragment structure features of various herbal medicines could be completely captured within 10-15 s,with minimal sample(＜0.5 mg)and solvent consumption(＜20 μL for one sample).Furthermore,a rapid differentiation and authentication strategy for TCMs based on H-oEESI-MS was proposed,including metabolic profile characterization,characteristic marker screening and identification,and multivariate statistical analysis model validation.In an analysis of 52 batches of seven types of Aconitum medicinal materials,20 and 21 key compounds were screened out as the characteristic markers of raw and processed Aconitum herbal medicines,respectively,and the possible structures of all the characteristic markers were comprehensively identified based on Com-pound Discoverer databases.Finally,multivariate statistical analysis showed that all the different types of herbal medicines were well differentiated and identified(R2X＞0.87,R2Y＞0.91,and Q2＞0.72),which further verified the feasibility and reliability of this comprehensive strategy for the rapid authentication of different TCMs based on H-oEESI-MS.In summary,this rapid authentication strategy realized the ultra-high-throughput,low-cost,and standardized detection of various complex TCMs for the first time,thereby demonstrating wide applicability and value for the development of quality standards for TCMs.