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目的:观察胶艾汤及其加味方参芪胶艾汤的植物雌激素样作用,并利用雌激素受体(ER)(+)MCF7细胞探讨其发挥雌激素样作用的可能靶点和机制。方法:正常昆明种性未成熟雌性小鼠60只,体重9~12 g,按体重均衡和随机的原则分为6组:正常组给予等体积蒸馏水,阳性药己烯雌酚组按0.35 mg.kg-1.d-1给予己烯雌酚,胶艾汤和参芪胶艾汤低,高剂量组分别按2.5,5.0 g.kg-1.d-1给予中药方剂,给药持续4 d。第5天眼眶取血,分离血清,称取子宫并计算子宫系数。通过MTT细胞增殖实验观察2个方剂含药血清对MCF7细胞增殖的影响;运用实时荧光定量RT-PCR技术检测含药血清对雌激素效应基因pS2及ERα,ERβ表达的调控作用;并以雌激素受体拮抗剂ICI182,780干预,探讨其发挥雌激素样作用的可能机制。结果:高剂量胶艾汤及参芪胶艾汤能够明显提高性未成熟小鼠的子宫系数(P<0.05);其含药血清能够显著促进MCF7细胞增殖(P<0.05或0.01),加入ER拮抗剂ICI182,780可抑制其促增殖作用(P<0.05或0.01)。与己烯雌酚组一致,2个方剂含药血清也可提高靶细胞pS2表达水平(P<0.01),加入ICI182,780可抑制pS2表达水平的提高(P<0.01);2个方剂含药血清还可明显提高ERα(P<0.05)和ERβ(P<0.01)的表达水平。结论:胶艾汤及参芪胶艾汤均具有一定的通过ER介导的雌激素样作用,并可提高靶细胞ER亚型(特别是ERβ)mRNA的表达水平。
OBJECTIVE: To observe the estrogen-like effect of Jia AI decoction and its additive Shenqi Jiaotai Decoction and to explore the possible target and mechanism of estrogen-like action by estrogen receptor (ER) (+) MCF7 cells. Methods: Sixty female Kunming normal immature female mice weighing 9-12 g were divided into 6 groups according to the principle of weight balance and randomization: the normal group was given equal volume of distilled water, and the positive drug diethylstilbestrol group was treated with 0.35 mg.kg-1 .d-1 given diethylstilbestrol, glue AI Tang and Astragalus AI Tang low, high-dose group were given 2.5,5.0 g.kg-1.d-1 prescription, dosing for 4 d. On the fifth day, the orbital blood was collected, the serum was separated, the uterus was weighed, and the uterine coefficient was calculated. The proliferation of MCF7 cells was observed by MTT cell proliferation assay. The expression of estrogen-responsive genes pS2, ERα and ERβ was detected by real-time fluorescent quantitative RT-PCR. Estrogen Receptor antagonist ICI182,780 interference, to explore its possible mechanism of estrogen-like role. Results: High-dose Jia AI Decoction and Shenqigao AI Decoction could significantly increase the uterine coefficient of sexual immature mice (P <0.05). The serum containing them can significantly promote the proliferation of MCF7 cells (P <0.05 or 0.01) The antagonist ICI182,780 inhibited its proliferation (P <0.05 or 0.01). Consistent with the diethylstilbestrol group, the serum containing 2 prescriptions also increased the expression level of pS2 in target cells (P <0.01), ICI182,780 inhibited the expression of pS2 (P <0.01) Significantly increased the expression of ERα (P <0.05) and ERβ (P <0.01). CONCLUSION: Jia AI Tang and Shen Qijiao AI Tang all have certain estrogen-like effects mediated by ER, and can increase the expression of ER subtypes (especially ERβ) mRNA in target cells.