四物汤基于雌激素受体亚型介导的雌激素样效应及其分子机理的研究
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摘要
目的:利用在体动物实验和体外雌激素受体(ER)阳性MCF7细胞培养实验研究四物汤的雌激素样效应及其分子机理。方法:40只未性成熟SD大鼠,体质量50 g,21 d,按照随机分组的原则分为4组:正常对照组、雌激素组、四物汤高剂量组和四物汤低剂量组。灌胃给药,2次/d,持续4 d后取材,称量子宫重量,计算子宫系数,并制备药理血清。用四物汤药理血清作用于MCF7细胞,并分别加入ERα和ERβ特异性拮抗剂MPP和PHTPP作为工具药,通过MTT法检测四物汤对MCF7细胞增殖的影响; Western blot法检测四物汤对MCF7细胞ERα、ERβ蛋白表达的影响。结果:四物汤可发挥雌激素样效应,增加未性成熟大鼠的子宫系数(P <0. 05或0. 01);四物汤药理血清可促进细胞增殖(P <0. 05或0. 01),该促增殖作用在加入MPP或PHTPP后均被减弱,且加入PHTPP后拮抗作用更显著(P <0. 01)。Western blot检测发现四物汤药理血清可提高MCF7细胞ERα、ERβ的表达水平,并可降低ERα与ERβ比值(P <0. 01)。结论:四物汤在整体动物水平可表现出雌激素样效应;体外实验进一步证实四物汤可通过ER通路,特别是ERβ途径影响ER阳性细胞的增殖活性,并可改变ERα与ERβ比值。
Objective: To study the estrogenic effects and its molecular mechanism of Siwu Decoction via in vivo animal experiment and in vitro ER( +) MCF7 cell test. Methods: Forty immature SD rats,weighing 50 g,21 days old,were randomly divided into 4 groups according to the principle of random distribution law: normal control group,estrogen group,high dose and low dose Siwu Dedoction groups. Gavage administration was given for 4 days and twice a day. The rats were sacrificed on the fifth day. The uterus was weighed and uterus coefficient was calculated. The blood was collected for the preparation of serum pharmacological research. The proliferation rate of MCF7 cells influenced by Siwu Decoction was determined by MTT assay with the application of ERα and ERβ antagonists MPP and PHTPP as tools. The protein expressions of ERα and ERβ in MCF7 cells affected by Siwu Decoction were detected by western blotting. Results: Siwu Decoction could increase the uterus coefficient of rats( P < 0. 05 or P <0. 01). Siwu Decoction could also promote MCF7 cell proliferation rate( P < 0. 05 or P < 0. 01) and it could be inhibited by MPP or PHTPP,especially by PHTPP( P < 0. 01). The pharmacological serum of Siwu Decoction could improve the protein expressionsof ERα and ERβ in MCF7 cells and reduce the ratio of ERα and ERβ( P < 0. 01). Conclusion: Siwu Decoction showed estrogenic effects in animal experiment. It could affect the proliferation of ER positive cells through ER,mainly ERβ pathway and change the ratio of ERα and ERβ.
Objective: To study the estrogenic effects and its molecular mechanism of Siwu Decoction via in vivo animal experiment and in vitro ER( +) MCF7 cell test. Methods: Forty immature SD rats,weighing 50 g,21 days old,were randomly divided into 4 groups according to the principle of random distribution law: normal control group,estrogen group,high dose and low dose Siwu Dedoction groups. Gavage administration was given for 4 days and twice a day. The rats were sacrificed on the fifth day. The uterus was weighed and uterus coefficient was calculated. The blood was collected for the preparation of serum pharmacological research. The proliferation rate of MCF7 cells influenced by Siwu Decoction was determined by MTT assay with the application of ERα and ERβ antagonists MPP and PHTPP as tools. The protein expressions of ERα and ERβ in MCF7 cells affected by Siwu Decoction were detected by western blotting. Results: Siwu Decoction could increase the uterus coefficient of rats( P < 0. 05 or P <0. 01). Siwu Decoction could also promote MCF7 cell proliferation rate( P < 0. 05 or P < 0. 01) and it could be inhibited by MPP or PHTPP,especially by PHTPP( P < 0. 01). The pharmacological serum of Siwu Decoction could improve the protein expressionsof ERα and ERβ in MCF7 cells and reduce the ratio of ERα and ERβ( P < 0. 01). Conclusion: Siwu Decoction showed estrogenic effects in animal experiment. It could affect the proliferation of ER positive cells through ER,mainly ERβ pathway and change the ratio of ERα and ERβ.
引文
[1]太平惠民和剂局.陈庆平,陈冰鸥,校注.太平惠民和剂局方[M].北京:中国中医药出版社,1996:207.
[2]郝庆秀,王继峰,牛建昭,等.四物汤及组方中药植物雌激素活性的实验研究[J].中华中医药学刊,2009,27(4):739-740.
[3]陈悦,冯放,裴育莹,等.四物汤治疗更年期综合征的作用机制研究[J].现代中西医结合杂志,2015,24(8):799-804.
[4]戴方伟,陈文文,毛栋森,等. SD雌性大鼠性发育早期性器官等脏器和性激素的动态变化[J].中国比较医学杂志,2009,19(7):35-36.
[5]杜惠兰.中西医结合妇产科学[M]. 2版.北京:中国中医药出版社,2012:25.
[6]李菡君.雌激素通过抑制P2Y2受体促进乳腺癌细胞的增殖作用[D].上海:第二军医大学,2010:28-30.
[7]赵锋,韩琳,伍会健.雌激素受体α的翻译后修饰与乳腺癌[J].生理科学进展,2012,43(3):229.
[8] Zhao Y,Deng C,Lu W,et al. let-7 microRNAs induce tamoxifen sensitivity by downregulation of estrogen receptor alpha signaling in breast cancer[J]. Mol Med,2011,17(11-12):1233-1241.
[9] Li G,Zhang J,Jin K,et al. Estrogen receptor-alpha36 is involved in development of acquired tamoxifen resistance via regulating the growth status switch in breast cancer cells[J]. Mol Oncol,2013,7(3):611-624.
[10]刘训碧,张绪良,王钢胜. ERβ在乳腺癌及癌旁组织中的表达及其意义[J].现代肿瘤医学,2017,25(14):2247-2248.
[11]刘瑞磊,陈晓萍,龙梅珺,等.乳腺癌发生发展与雌激素受体β的表达的关系[J].中华实验外科杂志,2015,32(10):2517-2518.
[12] Van Kruchten M,Van Der Marel P,De Munck L,et al. Hormone receptors as a marker of poor survival in epithelial ovarian cancer[J].Gynecol Oncol,2015,138(3):634-639.
[13] Bardin A,Boulle N,Lazennec G,et al. Loss of ERbeta expression as a common step in estrogen-dependent tumor progression[J]. Endocr Relat Cancer,2004,11(3):537-551.
[14] Cheng L,Li J,Han Y,et al. PES1 promotes breast cancer by differentially regulating ERαand ERβ[J]. J Clin Invest,2012,122(8):2857-2870.
[15]李魏玮.雌激素受体β对乳腺癌细胞耐药及增殖凋亡作用的研究[D].济南:山东大学,2014:6-9.
[16]陈风飞,高颖生,孙立,等.雌激素受体β在乳腺癌生长中的作用研究进展[J].药学进展,2017,41(1):65-70.
[17]孙佳琦,张甘霖,于明薇,等.植物雌激素与乳腺癌发病发展的研究进展[J].现代生物医学进展,2016,16(12):2368-2369.
[18]陈桂芝,陈长香.患病对围绝经期综合征及抑郁发生的影响[J].中国妇幼保保健,2011,26(10):1468-1470.
[19] Song X,Pan Z Z. Estrogen receptor-beta agonist diarylpropionitrile counteracts the estrogenic activity of estrogen receptor-alpha agonist propylpyrazole-triol in the mammary gland of ovariectomized Sprague Dawley rats[J]. J Steroid Biochem Mol Biol,2012,130(1-2):26-35.
[2]郝庆秀,王继峰,牛建昭,等.四物汤及组方中药植物雌激素活性的实验研究[J].中华中医药学刊,2009,27(4):739-740.
[3]陈悦,冯放,裴育莹,等.四物汤治疗更年期综合征的作用机制研究[J].现代中西医结合杂志,2015,24(8):799-804.
[4]戴方伟,陈文文,毛栋森,等. SD雌性大鼠性发育早期性器官等脏器和性激素的动态变化[J].中国比较医学杂志,2009,19(7):35-36.
[5]杜惠兰.中西医结合妇产科学[M]. 2版.北京:中国中医药出版社,2012:25.
[6]李菡君.雌激素通过抑制P2Y2受体促进乳腺癌细胞的增殖作用[D].上海:第二军医大学,2010:28-30.
[7]赵锋,韩琳,伍会健.雌激素受体α的翻译后修饰与乳腺癌[J].生理科学进展,2012,43(3):229.
[8] Zhao Y,Deng C,Lu W,et al. let-7 microRNAs induce tamoxifen sensitivity by downregulation of estrogen receptor alpha signaling in breast cancer[J]. Mol Med,2011,17(11-12):1233-1241.
[9] Li G,Zhang J,Jin K,et al. Estrogen receptor-alpha36 is involved in development of acquired tamoxifen resistance via regulating the growth status switch in breast cancer cells[J]. Mol Oncol,2013,7(3):611-624.
[10]刘训碧,张绪良,王钢胜. ERβ在乳腺癌及癌旁组织中的表达及其意义[J].现代肿瘤医学,2017,25(14):2247-2248.
[11]刘瑞磊,陈晓萍,龙梅珺,等.乳腺癌发生发展与雌激素受体β的表达的关系[J].中华实验外科杂志,2015,32(10):2517-2518.
[12] Van Kruchten M,Van Der Marel P,De Munck L,et al. Hormone receptors as a marker of poor survival in epithelial ovarian cancer[J].Gynecol Oncol,2015,138(3):634-639.
[13] Bardin A,Boulle N,Lazennec G,et al. Loss of ERbeta expression as a common step in estrogen-dependent tumor progression[J]. Endocr Relat Cancer,2004,11(3):537-551.
[14] Cheng L,Li J,Han Y,et al. PES1 promotes breast cancer by differentially regulating ERαand ERβ[J]. J Clin Invest,2012,122(8):2857-2870.
[15]李魏玮.雌激素受体β对乳腺癌细胞耐药及增殖凋亡作用的研究[D].济南:山东大学,2014:6-9.
[16]陈风飞,高颖生,孙立,等.雌激素受体β在乳腺癌生长中的作用研究进展[J].药学进展,2017,41(1):65-70.
[17]孙佳琦,张甘霖,于明薇,等.植物雌激素与乳腺癌发病发展的研究进展[J].现代生物医学进展,2016,16(12):2368-2369.
[18]陈桂芝,陈长香.患病对围绝经期综合征及抑郁发生的影响[J].中国妇幼保保健,2011,26(10):1468-1470.
[19] Song X,Pan Z Z. Estrogen receptor-beta agonist diarylpropionitrile counteracts the estrogenic activity of estrogen receptor-alpha agonist propylpyrazole-triol in the mammary gland of ovariectomized Sprague Dawley rats[J]. J Steroid Biochem Mol Biol,2012,130(1-2):26-35.
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