瘦素调节人卵巢黄素化颗粒细胞功能的体外研究

目的　探讨瘦素、促卵泡激素 (FSH)和胰岛素样生长因子Ⅰ (IGF Ⅰ )对人卵巢黄素化颗粒细胞雌二醇 (E2 )、孕酮 (P)生成的影响及可能的作用机制。方法　培养人黄素化颗粒细胞 ,分别以瘦素 (3 0ng/ml)、FSH (1 0ng/ml)、IGF Ⅰ (30 0ng/ml)以及相同终浓度的瘦素 +FSH、瘦素 +IGF Ⅰ、FSH +IGF Ⅰ、瘦素 +FSH +IGF Ⅰ对其刺激 2 4h ,对照组不加任何药物。对药物作用后的黄素化颗粒细胞行形态学观察、细胞计数 ;用放射免疫法检测培养液中E2 和P水平 ;同时用逆转录聚合酶链反应法对黄素化颗粒细胞行瘦素受体mRNA检测。结果　瘦素、FSH和IGF Ⅰ对黄素化颗粒细胞生长无影响。瘦素对黄素化颗粒细胞E2 生成量无影响 ,对FSH刺激E2 的生成也无影响 ,E2 水平作用前分别为 (0 10 3± 0 0 36 )pmol/10 0 0细胞、(0 32 3± 0 0 4 2 )pmol/10 0 0细胞 ,作用后分别为 (0 12 0± 0 0 0 8)pmol/10 0 0细胞、(0 343± 0 0 34)pmol/10 0 0细胞 ;而对IGF Ⅰ、FSH +IGF Ⅰ刺激黄素化颗粒细胞生成E2 有显著的抑制作用 ,E2 水平作用前后分别为 (0 318± 0 0 37)pmol/10 0 0细胞与 (0 4 93± 0 0 36 )pmol/10 0 0细胞、(0 193± 0 0 2 5 )pmol/10 0 0细胞与 (0 2 5 1± 0 0 33)pmol/10 0 0细胞 (P <0 0 5 ,<0     

催乳素及多巴胺对大鼠卵巢颗粒细胞产生甾体激素的影响

使用含有FSH和雄烯二酮的DME-F 12培养液培养未成熟大鼠的卵巢颗粒细胞,观察了不同剂量的催乳素和多巴胺对颗粒细胞合成孕酮和雌二醇的影响。结果表明,低浓度的催乳素(0.5 μg/ml)促进孕酮产生,高浓度时(5 μg/ml)抑制孕酮产生。催乳素能抑制雌二醇产生。低浓度的多巴胺(0.5 μg/ml)不影响这两种甾体激素的产生。当浓度为5μg/ml时能明显抑制这两种激素的产生。但是,多巴胺对颗粒细胞发挥作用的机制需进一步研究。     

卵巢黄素化颗粒细胞瘦素信号转导分子STAT3磷酸化在多囊卵巢综合征发病中的作用

目的:探讨瘦素在多囊卵巢综合征(PCOS)发病中的作用。方法:选择行IVF-ET治疗的30例PCOS患者根据体重指数(BMI)分为肥胖者(BMI≥24)15例和非肥胖者(BMI<24)15例,及同期排卵功能正常或单纯输卵管因素不孕的非PCOS肥胖者和非肥胖者各15例,采用ELISA检测各组血清瘦素水平;采用放射免疫法检测各组空腹血清胰岛素(FIN)水平;采用葡萄糖氧化酶法测定各组空腹血糖(FPG)水平,利用稳态模型(HOMA)计算胰岛素抵抗指数(即HOMA-IR);采用Western blotting检测卵巢黄素化颗粒细胞信号转导分子STAT3磷酸化水平;同法检测不同浓度的瘦素(0ng/ml、10ng/ml、100ng/ml、1000ng/ml)体外对正常人卵巢黄素化颗粒细胞STAT3磷酸化(p-STAT3)的影响。结果:①血清瘦素水平PCOS肥胖者最高,其后依次为对照组肥胖者、PCOS非肥胖者和对照组非肥胖者,各组之间两两比较,差异均具有显著性(P<0.05);②PCOS患者血清瘦素水平与BMI和HOMA-IR均呈显著正相关(r=0.707,P<0.01;r=0.761,P<0.01);③STAT3水平肥胖PCOS组最高,其后依次为肥胖对照组、非肥胖PCOS组和正常对照组,各组间两两比较,差异均具有显著性(P<0.05);④正常人卵巢黄素化颗粒细胞经不同浓度瘦素处理48h后,p-STAT3水平呈不同程度增加,至瘦素浓度达到100ng/ml时,p-STAT3水平达到高峰,随后呈下降趋势。结论:瘦素可能通过激活JAK2/STAT3信号传导通路参与PCOS排卵障碍的发生。     

卵巢颗粒膜细胞一氧化氮的分泌及激素调节作用

目的探讨排卵前人卵巢颗粒膜细胞一氧化氮（NO）的分泌与白细胞介素1β（IL-1β）的关系，及其调节雌、孕激素的作用。方法收集体外受精-胚胎移植时穿刺卵巢抽吸的卵泡液，取出卵细胞后，分离出颗粒膜细胞进行培养，分别进行向其内添加IL-1β及NO供体的实验，测定各自培养液中的NO、雌二醇(E2)、孕酮(P)量及芳香化酶的活性。结果添加IL-1β组颗粒膜细胞的NO分泌量为（537±23）μmol/L，不添加IL-1β组为（411±15）μmol/L（P<0.01）；添加NO供体S-硝基乙酰基青霉胺（SNAP）组颗粒膜细胞的E2分泌量为（1234±136）μg/L，不添加NO供体SNAP组为（1792±103）μg/L（P<0.01）；添加NO供体SNAP组颗粒膜细胞的P分泌量为（988±103）μg/L，不添加NO供体SNAP组为（2486±123）μg/L（P<0.001）；添加NO供体SNAP组颗粒膜细胞的芳香化酶活性为（0.422±0.052）Ci/ml，不添加NO供体SNAP组为（0.833±0.012）Ci/ml（P<0.01）。结论在排卵前，卵巢颗粒膜细胞可分泌NO并可能受IL-1β的调节，NO与IL-1β共同参与类固醇激素合成的调节。     

重组人生长分化因子-9蛋白对小鼠促排卵作用的研究

目的探讨重组蛋白生长分化因子-9(GDF-9)对小鼠卵泡发育的作用。方法选取30只性成熟雌性昆明小鼠随机分为3组,每组10只。对照组小鼠腹腔注射0.5 mL生理盐水;尿促性腺激素(hMG)组注射hMG 10 IU;联合组注射hMG 10 IU联合重组蛋白GDF-9 10μg;48 h后所有小鼠腹腔注射人绒毛膜促性腺激素(hCG)10 IU。注射hCG后16～20 h处死小鼠,比较各组小鼠卵巢指数(卵巢相对重量/相对体质量)及获卵数,酶联免疫吸附法(ELISA)法检测小鼠血清雌二醇(E2)含量,并对卵母细胞进行体外受精,观察各组受精情况。结果 hMG组、联合组获卵数(28.9±10.2、36.5±13.0)、卵巢指数[(32.26±6.33)mg/100 g、(38.80±8.90)mg/100 g]、血清E2含量[(45.64±14.19)ng/L、(52.02±16.08)ng/L]均高于对照组[11.4±5.4、(26.11±7.16)mg/100 g、(25.03±13.62)ng/L],差异有统计学意义(P=0.001、P=0.034、P=0.029;P=0.000、P=0.028、P=0.014);尤以联合组获卵数最多、卵巢指数最大、E2含量最高,与hMG组相比差异亦有统计学意义(P=0.025、P=0.031、P=0.043);对照组、hMG组、联合组组间小鼠卵母细胞体外受精率相近,差异无统计学意义(P0.05)。结论重组蛋白GDF-9可以辅助促性腺激素(Gn)有效增加小鼠促排卵获卵数,且对卵母细胞的正常受精能力无影响。     

促性腺激素释放激素激动剂长、短方案在体外受精-胚胎移植中的比较性研究

目的　探讨促性腺激素释放激素激动剂（GnRH-a）长、短方案控制性超排卵在体外受精-胚胎移植（IVF-ET）中的疗效。方法　将2000年1～5月进行IVF和单精子卵胞浆注射（ICSI）助孕的不孕患者，按病历奇、偶数编号分为GnRH-a长方案组（55例）和短方案组（54例）。长方案组从使用促性腺激素（Gn）治疗周期前的黄体中期开始使用GnRH-a0.9mg/d，至垂体完全降调节后，加用Gn；短方案组从月经第2天开始使用GnRH-a0.45mg/d，同时加用Gn。两组均在优势卵泡达18mm时，肌内注射人绒毛膜促性腺激素（hCG），36h后取卵行IVF及ICSI。结果　长方案组较短方案组，使用Gn前血清促卵泡激素（FSH）和黄体生成素（LH）水平降低［（4.4±1.2）IU/L比（6.3±1.7）IU/L,（2.7±1.5）IU/L比（4.4±2.8）IU/L,P<0.01］；注射hCG前血清雌二醇（E2）和LH水平降低［（7119±3584）pmol/L比（9523±3587）pmol/L，（1.0±1.0）IU/L比（4.0±3.4）IU/L,P<0.01］；每个卵子E2水平降低［（610±315）pmol/L比（935±450）pmol/L,P<0.01］；Gn用量增多［（28.0±8.6）支比（23.4±8.7）支,P<0.01］，用药时间增长［（11.1±1.2）d比（10.1±1.5）d,P<0.01］；两组平均获卵数、第2次成熟分裂中期卵子数、受精卵数、卵裂数、优质胚胎数及妊娠率无显著差异。结论　在IVF-ET，GnRH-a长、短方案能获得相同的控制性超排卵效果，且GnRH-a短方案能减少Gn用量和缩短治疗时间。     

抗孕激素RU486和ZK98734对双池培养的颗粒细胞和内泡膜细胞分泌功能的影响

本文观察了RU486和ZK98734对培养在羊膜双池培养系统中的猪卵巢颗粒细胞和内泡膜细胞在甾体激素生成过程中的影响。生长在羊膜两书侧的颗粒细胞和内泡膜细胞在加入或不加FSH、LH及不同浓度RU486或ZK98734的条件下培养48h。用RIA测定内、外池培养液中的孕酮（P）和雌二醇（E2）的浓度并与单独培养的结果相比较，同时亦与大鼠颗粒细胞的结果作比较。结果表明：1．有FSH刺激时，两种抗孕激素均明显抑制双池培养中颗粒细胞的P和E2产量；2．不论有或无LH刺激，两种抗孕激素均显著抑制双池培养中内泡膜细胞的P产量；3．双池培养系统模拟了两种卵巢细胞在体内的旁分泌调节关系，比单独培养更加合理；4与大鼠相比，猪卵巢细胞体外培养是研究避孕药对卵巢功能影响的适合模型。     

PRL对体外培养的猪颗粒细胞中FSH受体亲和力及孕酮生成的作用

卵泡刺激索(FSH)对其受体(FSH-R)的作用是卵泡发育过程的中心环节。泌乳素(PRL)是促进细胞分化与增殖的生长调节因子,但PRL亦可通过抑制促性腺激素分泌而抑制卵巢功能。该研究目的为进一步确定PRL对颗粒细胞中FSH-R亲和力及孕酮(P)合成分泌的影响。从幼猪(p)卵巢非闭锁卵泡中提取未成熟颗粒细胞分成5组进行体外培养,即:空白对照组、生理浓度的羊(o)PRL组(1205ng/ml)、3个高浓度oPRL组(25,50,75ng/ml)。5组细胞培养4天,培养基中均加入纯化的pFSH(10ng/ml)。每2天更换培养基1次,保存于-20℃行孕酮放射     

生长激素在多囊卵巢综合征促排卵中的作用

目的　探讨生长激素(GH)在多囊卵巢综合征(PCOS)患者促排卵中的作用。方法　测定130例PCOS患者(PCOS组)及107例正常妇女(对照组)的血中生殖激素及GH和胰岛素样生长因子Ⅱ(IGF-Ⅱ)的基础水平，并应用GH辅助促排卵方案治疗7例对人绝经期促性腺激素(hMG)反应不良的PCOS患者，观察疗效。结果　PCOS患者血中GH水平明显降低，肥胖者更为明显，非肥胖与肥胖者分别为(2.50±1.33)μg/L及(1.04±0.47)μg/L,而对照组肥胖与非肥胖者分别为(2.95±1.49)μg/L、（5.30±2.26）μg/L（P均<0.05）;PCOS组肥胖者IGF-Ⅱ水平为（136±27）nmol/L，高于非肥胖者的（123±20）nmol/L，两者比较，差异有显著性（P<0.05）。应用GH辅助促排卵治疗，可以明显减少hMG用量1～12支，缩短hMG刺激时间3～12d，增加优势卵泡的数量。结论　PCOS患者存在GH分泌障碍，应用GH辅助促排卵可以提高排卵率。     

抗孕激素RU486和ZK98734对双池培养的颗粒细胞和内泡膜细胞分泌功能…

本文观察了ＲＵ４８６和ＺＫ９８７３４对培养在羊膜双池培养系统中的猪卵巢颗粒细胞和内泡膜细胞在甾体激素生成过程中的影响。生长在单膜两侧的颗粒细胞和内泡膜细胞在加入或不加ＦＳＨ、ＬＨ及不同浓度ＲＵ４８６或ＺＫ９８７３４的条件下培养４８ｈ。用ＲＩＡ测定内、外池培养液中的孕酮（Ｐ）和雌二醇（Ｅ２）的浓度并与单独培养的结果相比较，同时亦与大鼠颗粒细胞的结果作比较。结果表明：１．有ＦＳＨ刺激时，两种抗孕激素均     

Effects of leptin on basal and FSH stimulated steroidogenesis in human granulosa luteal cells

BACKGROUND: Body weight influences fertility and studies in mice have indicated that leptin is one of the mediators of this effect. Leptin is believed to centrally stimulate the hypothalamic-pituitary axis resulting in increased gonadotropin release. Moreover, leptin is present in follicular fluid and the receptor is expressed in the human ovary. The aim of this study was to evaluate the direct effect of leptin on cultured human granulosa cell steroidogenesis. METHODS: Granulosa cells were obtained in connection with IVF procedures, and then cultured in a serum-free medium containing androstenedione (1 microM) for a total of 4 days. After 2 days of culture the medium was changed and the hormones under study were added. We tested the effect of leptin (1, 20, 100 ng/ml) on basal, FSH (10-100 ng/ml), and FSH (10-100 ng/ml)+IGF-I (30 ng/ml) stimulated steroidogenesis. RESULTS: Leptin (20 ng/ml and 100 ng/ml) significantly reduced basal and FSH-stimulated estradiol secretion (p<0.05). Basal and FSH (10 and 30 ng/ml) stimulated progesterone production was significantly inhibited by leptin 20 ng/ml, whereas leptin 100 ng/ml significantly reduced basal but not FSH stimulated progesterone production. Finally, steroidogenesis stimulated by IGF-I alone and in combination with FSH was not influenced by leptin. CONCLUSION: These results suggest that leptin acts directly to inhibit basal and FSH stimulated estradiol and progesterone production in cultured human granulosa cells. This raises the possibility that high circulating leptin levels as seen in obese women may compromise fertility through peripheral mechanisms.     

雌孕激素对体外卵泡黄素化颗粒细胞分泌巨噬细胞集落刺激因子的影响

目的　探讨雌、孕激素对卵泡黄素化颗粒细胞 (颗粒细胞 )分泌巨噬细胞集落刺激因子(M CSF)的调节作用。方法　抽取接受卵母细胞单精子显微注射 (ICSI)的 12例妇女的颗粒细胞 ,在不同浓度 ( 0、1× 10 - 7、1× 10 - 6 、1× 10 - 5 、1× 10 - 4、1× 10 - 3 mol L)的雌二醇或孕酮作用下培养 72h ,采用酶联免疫吸附法测定颗粒细胞培养液中的M CSF含量。结果　颗粒细胞在雌二醇或孕酮为 0mol L(即无雌二醇或孕酮作用———基础状态 )的情况下培养 72h后 ,分泌一定量的雌二醇 [( 1 0 2± 0 2 3 )×10 - 7mol L]、孕酮 [( 1 0 0± 0 12 )× 10 - 5 mol L]和少量的M CSF[( 4 7± 15)ng L] ;在 1× 10 - 6 ～ 1× 10 - 3mol L浓度的雌二醇作用下 ,M CSF分泌增加 ,其分泌随雌二醇浓度的增加而增加 ,较基础状态分别增加 2 3、4 5、6 9和 7 9倍 ;在≥ 1× 10 - 6 mol L各浓度的雌二醇作用下 ,M CSF分泌与基础状态比较 ,差异均有显著性 (P均 <0 0 5) ;在 1× 10 - 7mol L浓度的雌二醇作用下 ,M CSF分泌与基础状态比较 ,差异无显著性 (P >0 0 5) ;在 1× 10 - 7～ 1× 10 - 3 mol L浓度的孕酮作用下 ,M CSF分泌与基础状态比较 ,差异均无显著性 (P >0 0 5)。结论　雌二醇能促进颗粒细胞M CSF的分泌 ,孕酮对颗     

巨噬细胞集落刺激因子对卵泡颗粒黄体细胞性激素分泌的影响及其受体的测定

目的　了解人卵泡颗粒黄体细胞中巨噬细胞集落刺激因子 (M CSF)受体的表达 ,M CSF在体外对人卵泡颗粒黄体细胞产生雌、孕激素的影响。方法　采用免疫细胞化学染色法 ,测定 2 0例行卵母细胞浆内单精子注射治疗患者的卵泡颗粒黄体细胞的M CSF受体 ;采用酶免疫分析法 ,测定卵泡颗粒黄体细胞在M CSF(浓度分别为 0、10、2 5、5 0、10 0、2 5 0ng/ml)单独及与促卵泡激素 (FSH ,浓度75IU/ml)联合作用 72h后的上清液中雌二醇 (E2 )和孕酮 (P)的浓度。结果　约 80 %的颗粒黄体细胞膜上M CSF受体阳性。在无FSH存在时 ,M CSF空白对照颗粒黄体细胞培养上清液中E2 浓度为(2 185± 189)pmol/L ,P浓度为 (315 7± 4 0 1)nmol/L ;M CSF为 10～ 10 0ng/ml时 ,E2 浓度在 (2 789± 36 5 )～ (42 82± 318)pmol/L之间 ,P浓度在 (42 5 6± 5 95 )～ (7789± 82 8)nmol/L之间。在有FSH作用时 ,M CSF空白对照的E2 和P浓度分别为 (5 0 4 5± 4 86 )pmol/L和 (86 6 7± 92 3)nmol/L ;M CSF为 10～ 10 0ng/ml时 ,E2 浓度在 (6 5 6 7± 6 73)～ (8373± 935 )pmol/L之间 ,P浓度在 (10 999± 985 )～ (14 990±115 8)nmol/L之间。M CSF促颗粒黄体细胞E2 和P分泌的作用随浓度增加而增强 (P均 <0 0 5 ) ;M CSF与FSH共同作用下颗粒黄体     

巨噬细胞集落刺激因子在卵泡黄素化颗粒细胞中的表达

目的　了解巨噬细胞集落刺激因子 (M CSF)在人卵泡黄素化颗粒细胞中的表达 ,探讨促卵泡激素 (FSH)在体外对人卵泡黄素化颗粒细胞合成分泌M CSF的调节作用。方法　取接受卵母细胞单精子显微注射治疗的 2 0例妇女的卵泡黄素化颗粒细胞 ,采用免疫细胞化学染色法测定黄素化颗粒细胞中M CSF的表达 ,并将黄素化颗粒细胞单独培养及在不同浓度的FSH(2、5、15、2 5IU/ml)作用下培养 72h ,采用酶联免疫吸附法测定黄素化颗粒细胞培养液中的M CSF含量。结果　约 80 %的黄素化颗粒细胞胞浆有M CSF表达 ;黄素化颗粒细胞在培养 72h后 ,基础状态下M CSF分泌量为(5 9± 13)ng/L ,浓度为 2、5、15、2 5IU/ml的FSH作用下M CSF分泌量分别为 (16 5± 32 )、(2 10± 5 8)、(2 99± 6 6 )、(390± 78)ng/L ,在不同浓度的FSH作用下黄素化颗粒细胞培养液中M CSF的含量与基础状态下相比 ,差异均有极显著性 (P <0 0 1)。结论　卵泡黄素化颗粒细胞能合成分泌M CSF ;FSH能促进黄素化颗粒细胞M CSF的分泌 ,FSH对卵泡发育和成熟的调节 ,可能部分通过M CSF及其受体通道完成     

Human chorionic gonadotropin stimulation of relaxin secretion by luteinized human granulosa cells.

OBJECTIVE: To determine the effect of human chorionic gonadotropin (hCG) on relaxin secretion by long-term cultures of luteinized human granulosa cells (GC). DESIGN: Luteinized human GC were collected from 10 women undergoing in vitro fertilization (IVF) cycles. Luteinized human GC from each woman were plated in replicate wells at 1 x 10(5) cells/well and exposed to medium 199 (GIBCO, Grand Island, NY), medium 199 with 1 IU/mL hCG, and/or medium 199 with 100 IU hCG/mL. Luteinized human GC were maintained for up to 40 days in culture. Spent media were changed every 2 days and assayed for relaxin and progesterone (P) at the conclusion of each experiment. SETTING: Tertiary care center. PATIENTS, PARTICIPANTS: Luteinized human GC were obtained from women undergoing controlled ovarian hyperstimulation for IVF with one of the following regimens: (1) clomiphene citrate with human menopausal gonadotropins (hMG); (2) hMG alone; or (3) hMG with leuprolide acetate. All women were less than 40 years of age, in good health, and were not taking medications other than those used in the ovulation-induction regimen. MAIN OUTCOME MEASURES: Levels of P and relaxin in spent media. RESULTS: Relaxin secretion by luteinized human GC was dependent on hCG stimulation and was detected only after a time lag in culture. After relaxin secretion was detected, it was maintained throughout the culture period (10 to 22 days). Luteinized human GC produced P immediately under both basal and stimulated conditions. Progesterone production continued throughout the culture period with hCG-stimulated cells producing significantly greater P after 4 to 8 days in culture. CONCLUSIONS: Luteinized human GC obtained at the time of oocyte retrieval secrete relaxin in response to hCG stimulation and secrete P under both basal and hCG-stimulated conditions, thereby serving as a model to explore luteal function and control.     

GnRH-a体外对人卵巢黄素化颗粒细胞雌二醇和孕酮分泌量的影响

目的 :观察促性腺素释放激素激动剂 (GnRH -a)在体外对人卵巢黄素化颗粒细胞雌二醇 (E2 )和孕酮 (P)分泌量的影响。方法 :培养人卵巢黄素化颗粒细胞 ,分别用终浓度为 1.0、10 .0、10 0 .0ng ml的GnRH -a刺激 ,同时设对照组 ,培养时间为 2、4、6d。用放射免疫法检测黄素化颗粒细胞E2 和P的分泌量。结果 :培养 2d ,GnRH -a中、高浓度组E2 和P分泌量分别为 (12 2± 37) %、(12 8± 2 4 ) % ;(143± 32 ) %、(137± 2 9) %对照组为10 0 % ,均显著高于对照组 (P <0 .0 5) ;低浓度组E2 和P分泌量与对照组差异无显著性。随着细胞培养天数的增加 ,GnRH -a中、高浓度组E2 分泌量明显低于对照组 ,高浓度组E2 分泌量与培养天数呈负相关 (r =- 0 .75,P <0 .0 5)。结论 :GnRH -a对黄素化颗粒细胞分泌甾体激素功能的影响随着作用时间和浓度的不同而变化     

The effect of leuprolide acetate on steroidogenesis by granulosa and theca cells in vitro

Enhancement of follicular response to controlled ovarian hyperstimulation for human in vitro fertilization (IVF) has been suggested following pretreatment with leuprolide acetate (LA). However, additional human menopausal gonadotropin (hMG) is required to achieve follicle maturity in the presence of LA. We studied the effect of LA on steroidogenesis of granulosa and theca cells in vitro. Human granulosa cells obtained from IVF follicular fluid aspirations were cultured for 14 days in the presence and absence of human chorionic gonadotropin (hCG). hCG significantly enhanced progesterone (P) and estradiol (E₂) production by the cells, however, the addition of LA in concentrations of 10, 100, and 1000 ng/ml had no effect. Porcine granulosa cells were cultured for 48 hr in the presence and absence of follicle stimulating hormone (FSH) with the addition of LA at the same doses. LA did not affect the FSH-induced increase in P production. Porcine theca cells were cultured for 48 hr in the presence and absence of hCG. The addition of LA did not affect androstenedione (A) production by these cells. We conclude that in this dynamic model in vitro, LA does not inhibit or stimulate P or E₂ production by granulosa or A production by theca cells.     

Angiotensin II (AII) modulation of steroidogenesis by luteinized granulosa cells in vitro

Purpose The purpose of the present study was to evaluate the effect of angiotensin II and its inhibitor, saralasin, on steroid production by luteinized human granulosa cells in vitro. Granulosa cells were obtained from follicular fluid aspirations from human in vitro fertilization. Cultures were established in supplemented Ham's F-10 medium. Human chorionic gonadotropin and angiotensin II were added to culture media and the effect on steroid production was measured. Results Human chorionic gonadotropin alone stimulated production of progesterone, estradiol, and testosterone. The addition of angiotensin II resulted in a dose-dependent increase in progesterone production (428% increase compared to baseline). No effect was seen on estradiol or testosterone. However, a large increase (700%) in estradiol was seen with the addition of the competitive inhibitor of angiotensin II, saralasin. Conclusion We conclude that angiotensin II modulates progesterone production by human luteinized granulosa cells in vitro. The observed enhancement of estradiol production by angiotensin blockade suggests a tonic inhibition of estradiol secretion by endogenous angiotensin II. Presented at the 36th Annual Meeting Society for Gynecologic Investigation, March, 1989, San Diego, California.