机构地区:[1]中山大学附属第二医院妇产科,广州510120
出 处:《中华妇产科杂志》2009年第3期191-195,共5页Chinese Journal of Obstetrics and Gynecology
基 金:“十一五”国家科技支撑计划(2007BA104800);广东省医学科研基金(B2007053、B2008042)
摘 要:目的探讨抗苗勒管激素(AMH)对卵巢黄素化颗粒细胞激素分泌和芳香酶P450mRNA表达的影响。方法采集2006年6—12月在中山大学附属第二医院进行体外受精-胚胎移植(IVF-ET)的10例不孕患者的卵巢黄素化颗粒细胞进行原代培养,按照加入AMH浓度的不同,将颗粒细胞分为A、B、C、D、E组及睾酮对照组、空白对照组,A~E组分别加入1、5、10、20、50μg/L的AMH及1×10^4mol/L睾酮,睾酮对照组加入1×10^-7mol/L睾酮,空白对照组仅加入培养基。于培养24、48、72h时分别检测各组细胞中的雌二醇水平;培养72h后各组均进行细胞计数,并采用RT—PCR技术检测B、C、D、E组及睾酮对照组细胞中芳香酶P450mRNA的表达水平。结果(1)培养24、48、72h后颗粒细胞中的雌二醇水平,A组分别为(8.529±0.381)×10^4、(10.977±0.436)×10^4、(13.309±0.506)×10^4pmol/L,B组分别为(7.027±0.276)×10^4、(9.167±0.300)×10^4、(10.794±0.555)×10^4pmo]/L,C组分别为(6.039±0.226)×10^4、(7.585±0.548)×10^4、(8.797±0.518)×10^4pmol/L,D组分别为(5.118±0.460)×10^4、(5.716±0.496)×10^4、(6.205±0.667)×10^4pmo]/L,E组分别为(4.932±0.148)×10^4、(5.323±0.184)×10^4、(5.629±0.212)×10^4pmol/L,各组分别与空白对照组[分别为(0.001±0.001)×10^4、(0.006±0.003)×10^4、(0.029±0.011)×10^4pmol/L]比较,差异均有统计学意义(P〈0.01);B、C、D、E组分别与睾酮对照组[分别为(8.418±0.569)×10^4、(10.841±0.689)×10^4、(13.301±0.637)×10^4pmol/L]比较,差异也均有统计学意义(P〈0.01);B组分别与C、D、E组比较,C组分别与D、E组比较,差异也均有统计学意义(P〈0.01);D组与E组之间比较,差异无统计学意义(P〉0.05)。A、B、C、D、E组及睾酮�Objective To investigate the effect of anti-Mtillerian hormone (AMH) on hormone secretion and P450 aromatase mRNA expression from cultured human luteinized granulosa cells. Methods Human luteinized granulose cells were derived from 10 patients treated by in vitro fertilization-embryo transplantation (IVF-ET) in the Second Affiliated Hospital of Sun Yat-sen University from June to December 2006. Granulose cells were divided into group A, B, C, D, E depending on different concentration of AMH, testosterone group and blank group. 1 ~ 10^-7mol/L testosterone and 1,5,10,20,50 μg/L AMH were added into the culture medium of group A,B,C,D and E. 1 × 10^-7mol/L testosterone was added into the culture medium of testosterone group while no other ingredient was added into the medium of blank group. Estrogen levels in supemates were measured at 24,48,72 hours after cell incubation. RT-PCR was performed to detect the P450 aromatase mRNA expression in group B, C, D, E and testosterone group at 72 hours after cell incubation. Results (1) Estrogen levels in supernates of granulose cell culture at 24,48,72 hours were (8.529±0.381) ×10^4, (10.977 ±0.436) ×10^4, (13.309 ±0.506)×10^4 pmol/L in group A, (7.027 ± 0. 276)×10^4, (9. 167 ±0. 300) ×10^4, ( 10. 794 ±0. 555 ) ×10^4pmol/L in group B, (6. 039±0. 226)×10^4, (7.585±0.548)×10^4, (8.797 ±0.518) ×10^4 pmol/L in group C, (5. 118 ±0.460) ×10^4, (5.716 ± 0. 496) ×10^4, (6. 205 ±0. 667)×10^4pmol/L in group D, (4. 932 -±0. 148)×10^4, (5. 323 ±0. 184)×10^4, (5.629±0.212)×10^4 pmol/L in group E. When compared with blank group [(0.001±0.001) ×10^4, (0. 006 ±0. 003) ×10^4, (0. 029 ±0. 011 ) ×10^4 pmoL/L], the statistical differences were observed in group A,B,C, D,E(P 〈 0. 01 ) ; when compared with testosterone group [ (8. 418 ± 0. 569 ) ×10^4, ( 10. 841 ± 0. 689)×10^4, ( 13. 301 ±0. 637)×10^4pmol/L] , the statistical differences were observed i
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