ABSTRACT
Objective: To study the effective substance foundation of Ephedrae Herba and explore its mechanism, in order to further enrich the theory of drug resistance of Ephedrae Herba.Method: In this experiment, a compound model was used to establish rat model of Harmful Fluid Retention in upper Jiao. The Rats were randomly divided into model group, captopril group (4.38 mg·kg-1), Ephedrae Herba decoction group(468 mg·kg-1), polysaccharide group (265.36 mg·kg-1), volatile oil group (2.34 mg·kg-1), alkaloid group(40.71 mg·kg-1) and phenolic acid group (210.60 mg·kg-1), and normal group (10 mL·kg-1). The normal group and the model group were given the same volume of normal saline for four weeks. The 24 h urine volume of rats was collected by metabolic cage method. The changes of heart and lung tissue morphology were observed under light microscope. The heart index, lung index, left ventricular ejection fraction(LVEF), left ventricular short axis shortening rate(LVFS) and pulmonary permeability index, number(LPI), lung dry-wet ratio(W/D), creatine kinase isoenzyme(CK-MB), angiotensin Ⅱ(Ang Ⅱ), aldosterone(ALD), cardiac aquaporin 1(AQP1), lung AQP1, aquaporin-3(AQP3) and kidney AQP1, aquaporin-2(AQP2), interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) change were detected.Result: Compared with the normal group, heart and lungs of the model group were significantly damaged. The amount of 24 h urine, LVEF, LVFS of model rats were significantly reduced(Pα were significantly increased(PPα were significantly increased (PPα were significantly reduced (PPConclusion: Alkaloid components "Wen" and "Xin" are the effective substance basis of its action. The mechanism may be related to the inhibition of renin angiotensin aldosterone system (RAAS) and the anti-inflammatory effect.
ABSTRACT
This study offers preliminary insight into the phytoestrogen activity and mechanism of rehmapicrogenin. In this study, we characterized the estrogenic activity of rehmapicrogenin using immature female mice in vivo and MCF-7 cell proliferation assay in vitro. All the procedures for the care of the mice were conducted in accordance with the Regulations of Experimental Animal Administration issued by the State Committee of Science and Technology of the People’s Republic of China. Uterine wet weight/body mass ratios, Western blot assay for estrogen receptor, and serum estrogen levels of estradiol (E2), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were investigated. The effects of rehmapicrogenin, and the estrogen receptor antagonist ICI182,780, the estrogen receptor alpha antagonist MPP, the estrogen receptor beta antagonist THC, the G-protein coupled receptor 30 antagonist G15 combined with rehmapicrogenin on cell proliferation were examined in MCF-7 cells. Rehmapicrogenin (50 mg·kg-1) treatments demonstrated significant estrogenic activity by promoting the development of uterus in immature female mice, as well as increasing the expression of estrogen receptor alpha (ERα) and G-protein coupled receptor 30 (GPR30) at the protein level in uterus, and decreasing FSH and LH compared with the control group. Meanwhile, rehmapicrogenin (6 and 8 μmol·L-1) promoted the proliferation of MCF-7 cells, which were significantly antagonized by ICI182,780, MPP and G15. This study demonstrates rehmapicrogenin exerts estrogenic effects through ERα and GPR30.
ABSTRACT
Aim To evaluate the estrogen-like activity of Semen Descurainiae aqueous extracts (SD-ae), to deter-mine its effective chemical separation components and to study the mechanisms. Methods The estrogen-like ac-tivity of SD-ae and its effective chemical separation com-ponents were evaluated by the animal experiment, uterine weight test and cell experiment, namely E-SCREEN ex-periment. Estrogen receptor antagonist ICI182,780 inter-vention blocking experiment was carried out to detect the pathway of estrogen-like action; the HEK293 cells were co-transfected with the report gene carrier and the ERα, ERβ expression vector by cationic liposome, the report gene carrier was constructed via the estrogen-responsive component (ERE) and the report gene luciferase (Luc), then the estrogen-like signaling pathway was evaluated with standardized Luc activity; the expression of estrogen receptor ERα, ERβ and estrogen-induced gene PR mR-NA was detected by real-time fluorescence quantitative PCR. Results Compared with normal control, SD-ae low and high dose could significantly improve the uterine coefficient of immature female mice(P<0.05), and the oligosaccharides composition of Semen Descurainiae aque-ous extracts(SD-ae-Oli) and the polysaccharide composi-tion of Semen Descurainiae aqueous extracts(SD-ae-Pol) also significantly improved the uterine coefficient of im-mature female mice (P<0.01 or P<0.05); SD-ae, SD-ae-Oli and SD-ae-Pol had a significant proliferative effect on MCF-7 cells ( P <0.01 or P <0.05), while ICI182,780 intervened to block its proliferative effect. The reporter gene technology showed that the standardized Luc activities of SD-ae, SD-ae-Oli and SD-ae-Pol were significantly higher than those of the normal control when they were induced by ERβ respectively (P<0.01); and the SD-ae significantly increased the expression of ERβ mRNA in mouse uterus than the normal control, but no effect was found on the expression of ERα and PR mR- NA. Conclusions The estrogenic effect of SD-ae may be found at the first time, and its effective chemical sepa-ration components are SD-ae-Oli and SD-ae-Pol. Their estrogenic effects are mediated by ERβ. The molecular mechanism of the estrogenic effects is probably that SD-ae promotes the expression of ERβ mRNA.