Effects of Progestin and Antiprogestin on the Expression of FSH Receptor and LH Receptor mRNA in Porcine Granulosa and Thecal Cells

We havereported thattheprogestin and antiprogestin both inhibited secretion ofsteroid by porcine granulosa and thecal cells cultured in a dual chambersystem.Theprevious results suggested that this inhibition seemed to be via reduction of go-nadotropin stimulation[1 ,2 ] . The anterior pituitary cell types are responsible for thesynthesis and secretion of LH and FSH known as gonadotropes. LH and FSH exerttheir effects through activation of gonadotropin receptors present on the plasmamembran…     

米非司酮体内增强顺铂对人卵巢癌细胞的抗癌作用

目的观察米非司酮在体内对顺铂抗人卵巢癌细胞作用的影响。方法采用人卵巢腺癌细胞耐药亚株COC1/ 顺铂,建立人类卵巢癌裸鼠异种移植瘤耐药模型,观察体内应用米非司酮后荷瘤裸鼠肿瘤体积及其瘤组织形态学变化。应用RT-PCR技术检测瘤组织葡萄糖神经酰胺合成酶(GcS)mRNA表达。结果顺铂与米非司酮联用后,细胞形态发生变化,抑瘤率为70.48%,显著高于顺铂组的抑瘤率21.55%(P<0.01)。联合用药组和米非司酮组的移植瘤GcS mRNA表达较对照组显著下降,联合用药组尤为明显。结论无毒性剂量米非司酮可以在体内增加COC1/顺铂细胞对顺铂的敏感性,作用机制与抑制GcS mRNA表达有关。     

米非司酮体内增强顺铂对人卵巢癌细胞的抗癌作用

目的：观察米非司酮在体内对顺铂抗人卵巢癌细胞作用的影响。方法：采用人卵巢腺癌细胞耐药亚株COC1／顺铂，建立人类卵巢癌裸鼠异种移植瘤耐药模型，观察体内应用米非司酮后荷瘤裸鼠肿瘤体积及其瘤组织形态学变化，应用RT－PCR技术检测瘤组织葡萄糖神经酰胺合成酶（GcS)mRNA表达。结果：顺铂与米非司酮联用后，细胞形态发生变化，抑瘤率为70．48％，显著高于顺铂组的抑瘤率21．55％（P＜0．01），联合用药组和米非司酮组的移植瘤GcSmRNA表达较对照组显著下降，联合用药组尤为明显，结论：无毒性剂量米非司酮可以在体内增加COC1／顺铂细胞对顺铂的敏感性，作用机制与抑制GcSmRNA表达有关。     

体外颗粒细胞培养中抗苗勒管激素与卵泡刺激素受体mRNA表达的相关性

目的体外培养小鼠原代颗粒细胞培养液中,加入不同浓度的抗苗勒管激素(AMH),比较各组间小鼠颗粒细胞上卵泡刺激素受体(FSHR)mRNA的表达情况,探讨体外培养颗粒细胞中AMH与FSHR mRNA表达的关系。方法选取4～5周的昆明雌性小鼠,腹腔注射孕马血清促性腺激素,48～54 h后颈椎脱臼法处死小鼠,取双侧卵巢,采用机械分离法释放颗粒细胞,胰蛋白酶消化细胞,低速离心分离细胞,用含15%胎牛血清的DMEM/F12培养基置于37℃、5%CO2培养箱中培养。显微镜观察细胞形态,流式细胞术分离细胞,并用免疫荧光法对所培养的颗粒细胞进行鉴定。同时在体外培养的小鼠原代颗粒细胞培养液中加入不同浓度(5、10、20、30、40ng/mL)的AMH,测定每组培养基中AMH的终浓度,并用荧光定量PCR技术检测每组颗粒细胞中FSHR mRNA的表达。结果分离培养的颗粒细胞纯度大于80%;随AMH浓度加大,小鼠颗粒细胞中FSHR mRNA的表达逐渐降低,当AMH浓度为5和10 ng/mL时,与对照组比较差异有统计学意义(P<0.05),且5 ng/mL浓度组颗粒细胞上FSHR mRNA的表达量最高。结论用机械分离加胰酶消化加低速离心能得到纯度较高活性很好的颗粒细胞,用FSHR鉴定颗粒细胞是一种简便易行的方法,AMH浓度与颗粒细胞上FSHR mRNA表达呈一定负相关,表明AMH有可能通过抑制FSHR表达降低FSH敏感性,从而参与卵巢早衰的病理生理过程。     

大鼠卵泡发育过程中基质金属蛋白酶-2，-9的作用

目的：探讨基质金属蛋白酶-2,-9（MMP-2,-9）在大鼠卵泡发育的组织重建中的作用。方法：采用原位杂方法检测动情周期大鼠卵泡中MMP-2,-9mRNAs的基因表达变化,并通过密度扫描方法对原位杂交结果进行半定量分析。结果：在动情周期中,各情期卵巢中卵泡的颗粒细胞和发育良好的卵泡的膜细胞中均可检测到MMP-2m-RNA的杂交信号;MMP-9mRNA的表达始于已发育良好的窦前卵泡的膜细胞,此后在小窦状卵泡、窦状卵泡及排卵前卵泡的膜细胞中均检测到MMP-9mRNA的杂交信号。结论：MMP-2,-9可能通过其基因的表达变化参与卵泡的发育过程。     

Effect of Compound Recipe Gengniankang (更年康) on Senile Sexual Hormone and Expression of Estrogen Receptor in Bone of Climacteric Female Rats

Estrogenis a vital hormone to women's wholelife.It is aninevitable process that thelevel of ser-umestrogen decreases when women enter the agedphase,and that the ovary function is exhausted.Accompanying this process,theincidence of osteo-porosis is increased and affects the quality of lifeseverely.Using hormone replacement treat ment toprevent and treat osteoporosis is very effective.Itis proved that osteoporosis is related to the lack ofestrogen,which plays a veryi mportant role duringthe proc…     

Multifactor Regulation on Expressions of MMP-9 and TIMP-1 in Endometrial Stromal Cells

Recent evidence suggests that preparation of the endometrium for implantation is notmerely a question of adequate hormonal stimulation but that implantation also depends on theinteraction between the blastocyst and the endometrium and is mediated by cytokines,growthfactors,and adhesion molecules,which are produced and secreted by the endometrium andthe blastocyst[1].Implantation is a complex process that involves embryo apposition andattachment to the maternal endometrial epithelium,the extrace…     

米非司酮对大鼠黄体和子宫内膜纤溶酶原激活因子的调节作用

采用纤维蛋白铺盖和放射免疫方法研究了米非司酮(RU486)对培养颗粒细胞及黄体细胞纤溶酶原激活因子(包括组织型tPA和尿激酶型uPA)的作用。结果示RU486能明显拮抗入绒毛膜促性腺激素(hCG)促进颗粒细胞tPA活性及孕酮分泌的作用。RU486和PGF2α能明显促进黄体细胞tPA分泌,但孕酮产生明显被抑制。用组织学培养的方法,结果表明,RU486能刺激妊娠子宫内膜tPA和uPA分泌。这些结果表明RU486的抗生育作用部分能通过PA起到抗排卵、溶黄体及抗早孕的作用。     

孕酮拮抗剂米非司酮对子宫肌瘤的影响

给予子宫肌瘤患者（１８例）孕激素拮抗剂米非司酮（ＲＵ４８６）口服治疗（２５ｍｇ／ｄ），连续服药３个月。服药前后比较，肿瘤体积平均缩小３６．９％。卵泡刺激素（ＦＳＨ）、黄体生成素（ＬＨ）、雌二醇（Ｅ２）治疗前后无明显变化，而孕酮（Ｐ）明显降低。有出血病史的患者，治疗１周内出血停止，全部患者都发生闭经。研究结果提示：米非司酮口服可作为子宫肌瘤患者的辅助治疗方法     

EFFECT OF RU486 ON THE LUTEAL FUNCTION IN THE PREGNANT MOUSE

The effect of RU486, an antiprogestin, on luteal functions in the post-implantation mouse was studied. (1) In vivo experiment: Female adult mice were treated with RU486 at an oral dose of 100 mg/kg on day 1 and day 2 of pregnancy. Autopsy was performed     

子宫腺肌病与盆腔子宫内膜异位症患者性激素受体研究

目的 :探讨子宫腺肌病 ,盆腔子宫内膜异位症异位内膜中雌激素受体 (ER)、孕激受体 (PR)、雄激素受体(AR)、卵泡刺激素受体 (FSH -R)及黄体生成素受体 (LH -R)分布的差异。方法 :对子宫腺肌病 ,卵巢内膜异位症囊肿各 30例和 2 0份上述病例周期在位内膜 ,应用免疫组化S -P ,测定ER、PR、AR、FSH -R、LH -R ,并进行比较。结果 :子宫腺肌病异位内膜中AR、LH -R阳性率明显高于卵巢内膜异位囊肿异位内膜 (P <0 .0 5 ) ,而FSH -R阳性率显著低于卵巢内膜异位囊肿异位内膜 (P <0 .0 5 )。结论 :子宫腺肌病与卵巢内膜异位囊肿异位内膜AR、LH -R及FSH -R分布不同 ,可能是子宫腺肌病与卵巢内膜异位囊肿药物治疗疗效不同的原因所在。     

Effects of Antisense Oligodeoxynucleotide to Follicle-stimulating Hormone Receptor on the Expression of Proliferating Cell Nuclear Antigen and Vascular Endothelial Growth Factor in Primary Culture Cells Derived from Human Ovarian Mucinous Cystadenocarcino

The effects of antisense oligodeoxynucleotide (antisense ODN) to follicle-stimulating hormone receptor (FSHR) and follicle-stimulating hormone (FSH) on the expression of proliferating cell nuclear antigen (PCNA) and vascular endothelial growth factor (VEGF) were studied in primary culture cells derived from human ovarian mucinous cystadenocarcinoma (OMC). The prlmary OMC cells were cultured with the enzyme digestion method, and the expression of pan Keratin protein and FSHR mRNA was detected for identification of the cells. OMC cells were co-cultured with antisense ODN, nonsense ODN and FSH with different concentrations for 48 h and 72 h. The expression of PCNA and VEGF was detected by using SP immunohistochemistry. Compared with that in the control group, the PCNA and VEGF expression was increased obviously in FSH groups (P＜0.05 or P＜ 0.01), while decreased significantly in antisense ODN groups (P＜0. 05 or P＜0.01) and unchanged in nonsense ODN groups, respectively. Meanwhile, antisense ODN could antagonize the increased expression of PCNA and VEGF caused by FSH significantly (P＜0.01). It was suggested that FSH might promotethe development of OMC to some extent. Antisense ODN could inhibit the proliferative activity of OMC cells and the promoting proliferative activity enhanced by FSH.     

Effects of Parental Dietary Exposure to GMRice TT51 on the Male Reproductive System of Rat Off spring

ObjectiveTo evaluatethehealth effects of parental dietary exposure to GM rice TT51 on the male reproductive system of rat offspring.
MethodsRice-based diets, containing 60% ordinary grocery rice, MingHui63, or TT51 by weight, were given to parental rats (15 males/30 females each group) for 70 days prior mating and throughout pregnancy and lactation. After weaning, eightmale offspringratswere randomly selected at each group and fed with dietscorrespondent to their parents’ for 70 days. The effects of exposure to TT51 on male reproductive system of offspring rats were assessed through sperm parameters, testicular function enzyme activities, serumhormones (FSH, LH, and testosterone levels), testis histopathological examination, and the relativeexpression levels of selected genes along the hypothalamic-pituitary-testicular (HPT) axis.
ResultsNo significant differences were observed in body weight, food intake, organ/body weights, serum hormone, sperm parameters, testis function enzyme ACP, LDH, and SDH activities, testis histopathological changes, and relative mRNA expression levels of GnRH-R, FSH-R, LH-R, and AR along the HPT axis.
ConclusionThe resultsof this studydemonstrate that parental dietary exposure to TT51 reveals no significant differences on the reproductive system of male offspring rats compared with MingHui63 and control.     

Role of TGF-β1 in the Formation of Ovarian Interstitial Fibrosis in PCOS Rat

Objective To study the role of TGF-β1 in the formation of interstitial fibrosis and capsular thickening in the PCOS rats.
Methods PCOS rat model were established by subcutaneous injection DHEA. Estradiol （E2）, testosterone （T）, follicular stimulating hormone （FSH）, luteinizing hormone （LH）, LH/FSH and fasting insulin （FINS） hormone values of the model were tested by microparticle enzyme immunoassay, the pathohistology of PCOS rat ovaries was observed by HE staining, and their ultrastructure was tested by transmission electron microscope （TEM）. The expressions of TGF-β1 in PCOS group （n =20）, and control group（n=20） were detected and evaluated by immunohistochemistry and image analysis system.
Results The results of serum E2, T, LH/FSH, FINS, the ovarian pathological-histology and the ultrastructure showed that the model was established successfully. Compared with control group： there was no significant difference of TGF-β1 expression intensity in oocytes, granulosa cells of the different developing stages of follicle in PCOS （P〉0.05）. But it was markedly higher expressed in the theca cells of antral follicles （P〈0.01）, and stromal cells （P〈0.05） in PCOS group than in control group.
Conclusion The abnormal expression of TGF-β1 in PCOS is the main cause of capsular thickening and interstitial fibrosis. TGF-β1 also takes part in regulating PCOS follicle development and atresia.     

左旋18甲基炔诺酮和低剂量米非司酮用于紧急避孕的临床观察

观察左旋18甲基炔诺酮（LNG）和低剂量米非司酮（Ru486）用紧急避孕的有效性及可接受性。采用双盲随机比较方法吸收合格妇女100例,在一次无防护性交后72h内随机进入LNG组（50例）和Ru486组（50例）进行治疗。LNG组首次服用LNG0.75mg,12h重复1次;Ru486组首次服用Ru48610mg,12h服安慰剂一片,于预期下次月经日期的第7d进行随访,以了解避孕有效性及副反应。结果LNG组发生1例妊娠,失败率为2%。Ru486组无妊娠发生。避孕有效率两组分别为77.8%和100%（P<0.05）,有统计学差异。副反应发生率两组均在10%以内,且很轻微,月经正常者依次为90%和88%,结果证实左旋18甲基炔诺酮和低剂量米非司酮用于紧急避孕是安全有效的。     

含有RU486诱导系统的IL-2基因表达质粒的构建及功能鉴定

目的： 构建含有RU486诱导系统的小鼠白细胞介素2 (IL-2) 基因表达质粒,研究RU486系统对IL-2基因表达的调控作用。方法： 用限制性内切酶Cla Ⅰ处理含有RU486诱导系统的质粒pRS17和编码IL-2基因的质粒pUC57-IL-2,将IL-2基因插入到pRS17构建pRS-IL-2;通过PCR及限制性酶切鉴定重组质粒;将重组质粒体外转染SMMC-7721细胞,用不同浓度的RU486进行处理;将重组质粒通过水流动力学注射法注射到小鼠体内,腹腔注射RU486进行诱导;通过ELISA方法检测细胞上清及血清中IL-2基因的表达。结果： 重组质粒pRS-IL-2经过限制性内切酶消化及PCR分析,显示了预期的片段。细胞在1×10-8 mol?L-1的RU486诱导下,IL-2表达水平最高,是无RU486组的1.95倍（P<0.001）。注射质粒后,接受RU486诱导的小鼠血清中IL-2水平较诱导前升高320倍,而未接受RU486的小鼠血清未检测到IL-2的表达。结论：成功构建了含有RU486诱导系统的IL-2基因表达质粒,IL-2基因的表达依赖于诱导剂RU486的存在。     

益肾导痰调冲汤对多囊卵巢综合征大鼠瘦素及其受体和IGF-IR表达的影响

目的:观察益肾导痰调冲汤对多囊卵巢综合征(PCOS)大鼠瘦素及其受体和胰岛素样生长因子I受体(IGF-IR)表达的影响。方法:SD大鼠,采用"T+HCG"造模法建立PCOS大鼠模型。随机分为正常对照组、空白模型组、益肾导痰调冲汤等效剂量组(以下简称中药组)、西药达英-35组、达英加中药组、桂枝茯苓丸组各10只。观察卵巢组织形态学改变;放射免疫法测定血清内分泌水平;ELISA法测定瘦素(Lep);免疫组化法测定卵巢组织瘦素受体(OB-R)及IGF-IR表达。结果:益肾导痰调冲汤可改善PCOS大鼠卵巢组织结构,缩小卵巢体积,减少囊状卵泡及闭锁卵泡数目。降低LH、LH/FSH、T、Lep,增高E2水平,降低OB-R、IGF-IR阳性表达(P<0.05或P<0.01)。结论:益肾导痰调冲汤可改善PCOS大鼠lep与OB-R的结合障碍,调节OB-R、IGF-IR的生物利用度,从而改善PCOS大鼠的高雄激素血症及排卵障碍。     

促卵泡生成素,黄体生成素和胰岛素对多囊卵巢病人颗粒细胞甾 …

目的 研究在促卵泡生成素（ＦＳＨ）促黄体生成素（ＬＨ）和胰岛素（ＩＮＳ）刺激作用下多囊卵巢综合征（ＰＣＯＳ）病人与正常卵巢的颗粒细胞甾体激素产生的异同。方法 对１８例月经周期正常妇女卵泡期的３３个卵泡和８例ＰＣＯＳ的１００个肺泡的颗粒细胞行体外培养,利用放射免疫法测定培养液中的甾体激素水平,比较了两组颗粒细胞在ＦＳＨ,ＬＨ和ＩＮＳ刺激作用下下雌二醇（Ｅ２）和孕酮（Ｐ）的养液中的甾体激素水平,比较了