Genistein对雌性大鼠卵巢颗粒细胞FSHR基因表达的影响
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摘要
目的观察Genistein对雌性大鼠卵巢颗粒细胞卵泡刺激素受体(FSHR)基因表达的影响,及其与雌激素受体(ER)的关系。方法选取25～28日龄雌性大鼠,皮下注射孕马血清促性腺激素,建立促卵泡发育模型,分离卵巢颗粒细胞进行培养,分别给予Genistein(0、0.1、1、5、10、100μmol/L)及雌激素受体阻断剂ICI 182,780(1μmol/L)处理24 h后,采用逆转录-聚合酶链反应(RT-PCR)检测芳香化酶基因上游调控因子FSHR m RNA的表达。结果与空白对照组比较,10μmol/L Genistein组FSHR mRNA表达显著增加(P <0.05)。ICI 182,780(1μmol/L)预处理细胞30 min后加入1和10μmol/L Genistein,其FSHR mRNA表达分别较1和10μmol/L Genistein组显著下降(P <0.05或P <0.01)。结论 10μmol/L的Genistein可上调雌性大鼠卵巢颗粒细胞中FSHR表达,且该作用可能是通过ER介导的。
Objective To observe the effect of Genistein on the expression of follicle stimulating hormone receptor(FSHR) gene in ovarian granulosa cells of female rats and its relationship with estrogen receptor(ER). Methods Female rats aged 25-28 days were subcutaneously injected with pregnant mare serum gonadotrophin to establish follicular development model. Ovarian granulosa cells were isolated and cultured. Genistein(0, 0.1, 1,5, 10, 100 μmol/L) and estrogen receptor blocker ICI 182,780(1 μmol/L) were treated for 24 hours respectively. Reverse transcription-polymerase chain reaction(RT-PCR) was used to detect the expression of FSHR mRNA, an upstream regulator of aromatase gene.Results Compared with the blank control group, the expression of FSHR m RNA was significantly increased in the10 μmol/L Genistein group(P < 0.05). After ICI 182,780(1 μmol/L) pretreatment for 30 minutes, the expression of FSHR in ICI 182,780 pretreated cells with 1 and 10 μmol/L Genistein was significantly lower than that in the 1 and10 μmol/L Genistein groups(P < 0.05 or P < 0.01) respectively. Conclusion 10 μmol/L of Genistein can up-regulate the expression of FSHR in ovarian granulosa cells of female rats, and this effect may be mediated by ER.
Objective To observe the effect of Genistein on the expression of follicle stimulating hormone receptor(FSHR) gene in ovarian granulosa cells of female rats and its relationship with estrogen receptor(ER). Methods Female rats aged 25-28 days were subcutaneously injected with pregnant mare serum gonadotrophin to establish follicular development model. Ovarian granulosa cells were isolated and cultured. Genistein(0, 0.1, 1,5, 10, 100 μmol/L) and estrogen receptor blocker ICI 182,780(1 μmol/L) were treated for 24 hours respectively. Reverse transcription-polymerase chain reaction(RT-PCR) was used to detect the expression of FSHR mRNA, an upstream regulator of aromatase gene.Results Compared with the blank control group, the expression of FSHR m RNA was significantly increased in the10 μmol/L Genistein group(P < 0.05). After ICI 182,780(1 μmol/L) pretreatment for 30 minutes, the expression of FSHR in ICI 182,780 pretreated cells with 1 and 10 μmol/L Genistein was significantly lower than that in the 1 and10 μmol/L Genistein groups(P < 0.05 or P < 0.01) respectively. Conclusion 10 μmol/L of Genistein can up-regulate the expression of FSHR in ovarian granulosa cells of female rats, and this effect may be mediated by ER.
引文
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[14] Spagnuolo C,Russo GL,Orhan IE,et al. Genistein and cancer:current status, challenges,and future directions[J].Adv Nutr,2015,6(4):408-419.
[15]肖晓辉,赵薇,刘国艺,等.大豆异黄酮对衰老大鼠卵巢芳香化酶表达的影响[J].生理学报,2006,58(supple 1):178.
[16]张翼鸿,庞海燕,肖晓辉,等.大豆异黄酮及其主要活性成分genistein对大鼠卵巢雌激素受体α表达的影响[J].中华医学杂志,2011,91(28):1987-1991.
[17]庞海燕,孟毅,赵薇,等.大豆异黄酮及主要活性成分Genistein对大鼠卵巢LRH-1基因表达的影响[J].国际遗传学杂志,2012,35(3):151-156.
[18] Yin D,Zhu Y,Liu L,et al. Potential detrimental effect of soy isoflavones on testis sertoli cells[J]. Zhong Nan Da Xue Xue Bao Yi Xue Ban,2014,39(6):598-604.
[19] Xi YD,Zhang DD,Ding J,et al. Genistein Inhibits Aβ25-35-Induced Synaptic Toxicity and Regulates CaMKII/CREB Pathway in SH-SY5Y Cells[J]. Cell Mol Neurobiol,2016,36(7):1151-1159.
[20] Zhou Y,Xu JN,Zeng C,et al. Metformin Suppresses Prostaglandin E2-Induced Cytochrome P450 Aromatase Gene Expression and Activity via Stimulation of AMPActivated Protein Kinase in Human Endometriotic Stromal Cells[J]. Reprod Sci,2015,22(9):1162-1170.
[21]匡野,庞海燕,肖晓辉,等.大豆异黄酮对围绝经期大鼠卵巢FSHR表达的影响[J].中国中医药科技,2011,18(4):311-312,314.
[22] Smith HO,Arias-Pulido H,Kuo DY,et al. GPR30 predicts poor survival for ovarian cancer[J]. Gynecol Oncol,2009,114(3):465-471.
[2] Setchell KDR. The history and basic science development of soy isoflavones[J]. Menopause,2017,24(12):1338-1350.
[3] Takagi A,Kano M,Kaga C. Possibility of breast cancer prevention:use of soy isoflavones and fermented soy beverage produced using probiotics[J]. Int J Mol Sci,2015,6(5):10 907-10 920.
[4] Varinska L,Gal P,Mojzisova G,et al. Soy and breast cancer:focus on angiogenesis[J]. Int J Mol Sci,2015, 16(5):11728-11 749.
[5] Spagnuolo C,Russo GL,Orhan IE,et al. Genistein and cancer:current status, challenges, and future directions[J].Adv Nutr,2015,6(4):408-419.
[6] Siow RC,Mann GE. Dietary isoflavones and vascular protection:activation of cellular antioxidant defenses by SERMs or hormesis[J]. Mol Aspects Med,2010,31(6):468-477.
[7] Ye L,Chan MY,Leung LK. The soy isoflavone genistein induces estrogen synthesis in an extragonadal pathway[J].Mol Cell Endocrinol,2009,302(1):73-80.
[8] Casarini L,Moriondo V,Marino M,et al. FSHR polymorphism p.N680S mediates different responses to FSH in vitro[J]. Mol Cell Endocrinol,2014,393(1/2):83-91.
[9] Ghosh S,Lu Y,Hu Y. A Role of CREB in BRCA1 Constitutive Promoter Activity and Aromatase Basal Expression[J]. Int J Biomed Sci,2008,4(4):260-265.
[10] Samarajeewa NU,Docanto MM,Simpson ER,et al. CREBregulated transcription co-activator family stimulates promoterⅡ-driven aromatase expression in preadipocytes[J].Horm Cance,2013,4(4):233-241.
[11]温海霞,孟毅,杨永斌,等.Genistein对大鼠卵巢颗粒细胞CYP19表达的影响[J].中国医药导报,2016,13(35):20-23.
[12] Solomon LA,Ali S,Banerjee S,et al. Sensitization of ovarian cancer cells to cisplatin by genistein:the role of NF-kappaB[J]. J Ovarian Res,2008,1(1):9-15.
[13]李超,刘艾林,杜冠华.天然异黄酮类化合物的药理学研究进展[J].中国新药杂志,2013,22(12):1415-1420.
[14] Spagnuolo C,Russo GL,Orhan IE,et al. Genistein and cancer:current status, challenges,and future directions[J].Adv Nutr,2015,6(4):408-419.
[15]肖晓辉,赵薇,刘国艺,等.大豆异黄酮对衰老大鼠卵巢芳香化酶表达的影响[J].生理学报,2006,58(supple 1):178.
[16]张翼鸿,庞海燕,肖晓辉,等.大豆异黄酮及其主要活性成分genistein对大鼠卵巢雌激素受体α表达的影响[J].中华医学杂志,2011,91(28):1987-1991.
[17]庞海燕,孟毅,赵薇,等.大豆异黄酮及主要活性成分Genistein对大鼠卵巢LRH-1基因表达的影响[J].国际遗传学杂志,2012,35(3):151-156.
[18] Yin D,Zhu Y,Liu L,et al. Potential detrimental effect of soy isoflavones on testis sertoli cells[J]. Zhong Nan Da Xue Xue Bao Yi Xue Ban,2014,39(6):598-604.
[19] Xi YD,Zhang DD,Ding J,et al. Genistein Inhibits Aβ25-35-Induced Synaptic Toxicity and Regulates CaMKII/CREB Pathway in SH-SY5Y Cells[J]. Cell Mol Neurobiol,2016,36(7):1151-1159.
[20] Zhou Y,Xu JN,Zeng C,et al. Metformin Suppresses Prostaglandin E2-Induced Cytochrome P450 Aromatase Gene Expression and Activity via Stimulation of AMPActivated Protein Kinase in Human Endometriotic Stromal Cells[J]. Reprod Sci,2015,22(9):1162-1170.
[21]匡野,庞海燕,肖晓辉,等.大豆异黄酮对围绝经期大鼠卵巢FSHR表达的影响[J].中国中医药科技,2011,18(4):311-312,314.
[22] Smith HO,Arias-Pulido H,Kuo DY,et al. GPR30 predicts poor survival for ovarian cancer[J]. Gynecol Oncol,2009,114(3):465-471.