地黄苦苷元的雌激素样活性筛选研究

Evaluation of the estrogenic effects of rehmapicrogenin

探讨地黄苦苷元(rehmapicrogenin)的植物雌激素活性及其作用机制。采用小鼠子宫增重实验和MCF-7细胞增殖实验对地黄苦苷元进行雌激素样活性筛选。动物实验操作符合中华人民共和国"实验动物管理条例"。Western blot法检测子宫组织雌激素受体表达量, ELISA法检测小鼠血清雌二醇(E2)、黄体生成素(LH)、卵泡刺激素(FSH)水平的变化。采用特异性雌激素受体拮抗剂(ICI182,780)、雌激素受体α阻断剂(MPP)、雌激素受体β阻断剂(THC)、雌激素膜受体GPR30阻断剂(G15)进行MCF-7细胞增殖拮抗实验探讨其作用机制。结果表明,地黄苦苷元(50 mg·kg-1)显著提高性未成熟小鼠子宫指数(P<0.05),显著降低血清卵泡刺激素(FSH)、黄体生成素(LH)的水平(P<0.01);地黄苦苷元(6和8μmol·L-1)能极显著促进MCF-7细胞增殖(P<0.01),且其对MCF-7细胞的促增殖作用可被ICI182,780、MPP、G15拮抗; Western blot结果显示,地黄苦苷元能显著增加小鼠子宫ERα、GPR30表达。本研究结果表明地黄苦苷元具有雌激素样活性,且通过ERα、GPR30共同介导。

This study offers preliminary insight into the phytoestrogen activity and mechanism of rehmapi‐crogenin. 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.