抑制素B在正常子宫内膜及外周血中表达的相关性分析

目的研究正常子宫内膜组织INH亚单位的表达及分布情况以及外周血中INH—B水平以探讨INH在子宫内膜中的作用。方法选择2006年1月～12月就诊的输卵管因素不孕者的增殖期子宫内膜16例，宫颈癌患者之早分泌期（11例）和晚分泌期子宫内膜（17例）进行免疫组化检测。于月经第3～5天或经前第7天抽取外周血检测INH—B。所有病例术前半年未服任何激素类药物。结果INH三个亚单位在正常子宫内膜组织中表达，随月经周期的变化染色反应逐渐增强。在正常增殖期子宫内膜组织中BB亚单位的表达与血INH—B水平呈正相关（r=0．539，0．767，P〈0．05）。结论INH亚单位在正常子宫内膜组织中的表达及在外周血中表达的相关性说明了INH可能是自分泌和旁分泌信号的组成部分，在子宫内膜成熟过程中起重要作用。     

Induction of micronuclei in somatic cells of mice exposed to x-rays or nonylphenol and to a combination of both agents

The effects of X-rays, nonylphenol (NL) and combination of both agents on the induction of micronuclei in mouse somatic cells were investigated. Pzh: SFIS mice were exposed during 2 weeks, 5 days per week to X-rays (doses: 0.05 Gy, 0.10 Gy,0.20 Gy), nonylphenol (doses: 25 mg/kg bw NL, 50 mg/kg bw NL, 100 mg/kg bw NL) and to a combination of X-rays and nonylphenol (doses: 0.05 Gy + 25 mg/kg bw NL, 0.10 Gy + 50 mg/kg bw NL). Samples from peripheral blood were taken 1 week after the start of exposure and 24 h after the end of exposure, whereas samples from bone marrow were taken 24 h after the end of exposure. Results obtained show that ionizing radiation, nonylphenol and combination of X-rays-NL in low doses induced micronuclei in peripheral blood and bone marrow reticulocytes. In contrast combined exposure to higher doses of both agents caused reduction frequency of micronuclei in the comparison to effects of X-rays acting alone. In bone marrow polychromatic erythrocytes the induction of micronuclei was enhanced after combined exposure to both agents in lower and higher doses.     

邻苯二甲酸二丁酯雄性生殖毒性分子作用机制

目的研究邻苯二甲酸酯类对大鼠睾丸毒性作用的分子机制。方法运用RT-PCR方法观察邻苯二甲酸二丁酯（DBP）对大鼠子代不同发育阶段睾丸的苗勒氏管抑制物质（MIS）、雄激素结合蛋白（ABP）和抑制素（inhibin）表达水平的影响,并采用ELISA法检测DBP染毒后大鼠睾酮水平的变化情况。结果与对照组比较,各DBP染毒组大鼠睾丸中MIS表达差异无统计学意义,血清总睾酮水平差异无统计学意义,但高剂量DBP（1000 mg/kg）组睾丸支持细胞中ABP和抑制素（inhibin）的mRNA表达量显著减少,且与睾丸损害呈时间-效应关系。结论环境内分泌干扰物DBP可干扰大鼠发育过程中睾丸支持细胞ABP和inhibin的表达,其对性腺发育的影响可能是DBP雄性生殖毒性的主要作用机制之一。     

The biphasic modulation of inhibin mRNA levels and secretion by PMSG in rat granulosa cells in vitro

Granulosa cell cultures derived from diethylstilboestrol-treated immature rats were used to study the in vitro effect of pregnant mare serum gonadotrophin (PMSG) on steady state mRNA levels for the inhibin alpha and beta A subunits and the secretion of immunoreactive inhibin and progesterone. After 48 h treatment the dose-response curve of PMSG revealed a maximum stimulation (2.5-3.5 fold) of cytosolic alpha and beta A mRNAs over the range of 1 to 10 mU PMSG mL-1, with corresponding stimulation of inhibin secretion. A high dose of PMSG (160-500 mU mL-1) clearly suppressed inhibin alpha mRNA levels as well as inhibin secretion, whereas progesterone (P) was maximally stimulated (up to 600 fold). Although the level of cytosolic inhibin beta A subunit mRNA was also down-regulated by a high concentration of PMSG in the culture medium, the doses required to suppress its mRNA level to less than those of the control varied. These data demonstrate that low doses of follicle stimulating hormone/luteinizing hormone (FSH/LH)-like (PMSG) activity enhances and high doses decrease the steady-state mRNA levels of inhibin in rat granulosa cells in vitro; this biphasic regulation in vitro reflects the differential regulation of inhibin secretion observed during the rat oestrous cycle.     

氟对体外大鼠睾丸支持细胞雄激素结合蛋白和抑制素B mRNA表达的影响

目的研究氟对体外培养大鼠睾丸支持细胞雄激素结合蛋白(ABP)和抑制素B(INHB)转录水平的影响。方法建立支持细胞体外培养模型,以浓度分别为2.5、5.0、10.0和20.0mg/L氟化钠溶液染毒细胞,用RT-PCR方法检测ABP和INHB mRNA的相对表达量。结果①ABP mRNA相对表达量在各剂量组与对照组相比,2.5mg/L组高于对照组,且差异有统计学意义(P0.05);5.0mg/L高于对照组,但差异没有统计学意义;其余各组低于对照组,但差异没有统计学意义(P0.05)。②INHB mRNA相对表达量在各剂量组与对照组比较,2.5和5mg/L组均高于对照组,差异具有统计学意义(P0.05);其余两组低于对照组,但是差异均无统计学意义(P0.05)。结论在2.5～20.0mg/L剂量范围内,未观察到氟对体外培养大鼠睾丸支持细胞ABP和INHB mRNA的表达有明显影响。     

邻苯二甲酸二丁酯对大鼠睾丸支持细胞的影响

目的探讨邻苯二甲酸二丁酯(DBP)对睾丸支持细胞超微结构和功能的影响。方法将6周龄雄性SD大鼠随机分成0、250、500和1000mg/kg组,每组16只。给予DBP灌胃,分别于染毒2周和4周后各处死8只动物。采用放射免疫法测定血清卵泡刺激素,逆转录聚合酶链反应测定雄激素结合蛋白mRNA和抑制素αmRNA的相对表达量,用透射电镜观察支持细胞的超微结构。结果DBP染毒4周后,卵泡刺激素表现出明显的上升趋势,250和1000mg/kg组与对照组比较差异有统计学意义;精子头计数和每日精子生成量呈现出单一的下降趋势,500和1000mg/kg组与对照组比较差异有统计学意义。雄激素结合蛋白mRNA和抑制素αmRNA表达水平随着DBP染毒剂量的增加呈现出明显的下降趋势,无论是染毒2周还是4周,500和1000mg/kg组与对照组比较差异均有统计学意义。0、250、500和1000mg/kg组的雄激素结合蛋白mRNA相对表达量在染毒2周后分别为0.89±0.15、0.85±0.23、0.54±0.17和0.52±0.16,染毒4周后分别为0.76±0.16、0.73±0.17、0.47±0.14和0.38±0.14;抑制素αmRNA的相对表达量在染毒2周后分别为0.88±0.16、0.61±0.12、0.48±0.15和0.47±0.11,染毒4周后分别为0.75±0.19、0.56±0.16、0.53±0.08和0.45±0.10。电镜观察结果显示,1000mg/kg组支持细胞胞浆中初     

神经生长因子对子宫内膜基质细胞增殖及子宫腺肌病雌激素受体α表达的影响

目的:探讨神经生长因子(NGF)对离体培养在位子宫内膜基质细胞增殖及子宫腺肌病雌激素受体α表达的影响。方法:体外原代培养人子宫内膜细胞,用0 ng/ml、10 ng/ml、50 ng/ml和100 ng/ml的神经生长因子分别干预,用MTT法测定内膜基质细胞增殖情况,用Westen-blotting测定50 ng/ml NGF对基质细胞雌激素受体α表达的影响。结果:50 ng/ml的NGF可促进子宫内膜基质细胞增殖,细胞存活率为82.61%,比其他浓度NGF显著升高(P<0.05)。50 ng/ml NGF对病例组和对照组不同细胞雌激素受体表达有影响,病例组腺上皮细胞和基质细胞总体均数差异均具有统计学意义(P<0.05)。结论:50 ng/ml的NGF可促进在位内膜基质细胞增殖及子宫腺肌病腺上皮细胞和基质细胞雌激素受体α表达。     

Oral contraceptives prevent the development of endometriosis in the chicken chorioallantoic membrane model

BACKGROUND: Fundamental and genetic differences between women in the endometrium may cause some to develop endometriosis, whereas others do not. Oral contraceptives (OC) may have an effect on the endometrium, rendering the development of endometriosis less likely. STUDY DESIGN: Endometrium from women using OC (OCE) and menstrual endometrium (ME) from normal cycling women were transplanted onto the chicken chorioallantoic membrane (CAM), and endometriosis-like lesion formation was evaluated. Microarray gene expression profiling was performed to identify differentially expressed genes in the endometrium from these groups. Microarray data were validated by real-time PCR. RESULTS: Less endometriosis-like lesions were formed after transplantation of OCE than after transplantation of ME (p<.05). Most of the differentially expressed genes belong to the TGFbeta superfamily. Real-time PCR validation revealed that inhibin betaA (INHBA) expression was significantly decreased in OCE as compared to ME. CONCLUSION: OC use affects the characteristics of endometrium, rendering it less potent to develop into endometriosis.     

Citalopram as an Inhibitor of Voluntary Ethanol Intake in the Male Rat

Psychological dependence was induced in rats by a 1-year intermittent exposure to intoxicating doses of ethanol, and recorded by the rat’s ability to later take the same dose of ethanol independent of the offered concentration. Citalopram (10 or 40 mg/kg/day) was given for 3 weeks with ethanol available only the first and the last day; 10 mg/kg had no effect. On the first treatment day 40 mg/kg decreased ethanol intake. On the last treatment day 40 mg/kg had no effect. The following week the ethanol intake was higher than before the treatment in the 40 mg/kg group. During the four posttreatment weeks the ethanol intake of the 40 mg/kg group dropped significantly. Citalopram was retested 18 weeks after the first treatment during 1 week, with continuous access to ethanol; 10 mg/kg had no effect and 40 mg/kg decreased ethanol intake at day 1, reaching a minimum in day 3. A tolerance to this effect was seen at the end of the week. Thus, in this model an acute dose of citalopram can decrease ethanol intake, but tolerance to this effect develops when citalopram is given both with and without access to ethanol.     

Toxicity studies on tetramethylthiuram monosulfide

Tetramethylthiuram monosulfide (TMTM), when given in a small dose of 26 mg/kg po, caused prolongation of the hexobarbital sleeping time in female Wistar rats. This effect was enhanced by increasing the dose to 867 mg/kg po and was associated with a lengthening of the zoxazolamine paralysis time. Both effects are causally related to an inhibition of the microsomal monooxygenases. One of the reasons for this inhibition conceivably is the concomitant decrease in the conversion of palmitic acid (incorporation of ¹⁴C radioactivity) into the phospholipids of the microsomal membrane. In contrast, other clinicochemical parameters of liver function detectable in the blood remained normal even after oral doses of 26 mg/kg TMTM administered on 5 consecutive days per week over a period of 4 weeks. However, this treatment caused a decrease in the erythrocyte count and hemoglobin content in the blood, together with a reduction of the relative liver weight, body weight, and the intake of food, while the consumption of drinking water was increased. Histologically, there was evidence of hepatocellular and renal tubular swelling. In female mice given acute oral doses of TMTM, the LD₅₀ was calculated as 818 (583–995) mg/kg. In mutagenicity tests, TMTM caused point mutations in strains TA 100 and TA 1535 (Salmonella typhimurium LT 2). The results indicate that the toxic potential of TMTM is comparable to that of other thiurams. A tentative hygienic threshold limit of 5 mg/m³ is proposed for the working environment.     

Interception. IV: Failure of mifepristone (RU 486) as a monthly contragestive, "Lunarette"

The new antiprogestin mifepristone (RU 486) was studied as a contragestive for continuous fertility control in 24 menstrual cycles. Two pregnancies out of three conceptions continued in spite of antiprogestin treatment. To date, mifepristone at the doses used appears to be inadequate for monthly use.     

基质金属蛋白酶-7异常表达与子宫内膜癌发病关系的探讨

目的:探讨MMP-7与子宫内膜癌发病及转移的关系。方法:利用免疫组化的方法观察72例子宫内膜癌病变组织、64例内膜癌前病变组织和80例正常子宫内膜中MMP-7及其天然抑制物TIMP-1的表达情况。结果:在内膜癌组、内膜癌前病变组MMP-7的阳性表达分别为87.5%(63/72)、75%(48/64),明显高于正常内膜组43.8%(35/80)(P<0.01),而两组之间无显著差异。内膜癌组和癌前病变组TIMP-1的阳性表达分别为80.6%(58/72)、65.6%(42/64),也明显高于正常内膜组38.8%(31/80)(P<0.01)。随着内膜癌临床分期的增加,MMP-7的表达明显增强,Ⅱ期以上的内膜癌MMP-7表达均明显高于Ⅰ期(P<0.05)。在不同组织学分类中,组织分化程度低的内膜癌MMP-7表达明显高于组织分化程度高者。淋巴结转移阳性的内膜癌组织,MMP-7表达高于淋巴结转移阴性的内膜癌组织(P<0.01)。而TIMP-1则与上述病理学参数无显著相关性。结论:MMP-7在内膜癌发生过程中起到了重要作用,MMP-7表达增强和TIMP-1分泌相对不足,是内膜癌浸润和转移的重要促进因素。     

Human chorionic gonadotrophin raises serum immunoreactive inhibin levels in men with hypogonadotrophic hypogonadism

Inhibin is a gonadal glycoprotein hormone involved in the regulation of FSH. To elucidate the regulation of inhibin production we investigated the acute (daily for 1 week) and chronic (9-10 months of follow-up) changes in immunoreactive inhibin, testosterone, LH and FSH levels in the serum of three hypogonadotrophic hypogonadal patients treated first with hCG alone (for 3-6 months) and then hCG combined with FSH (1-5 months). One patient was unexpectedly resistant to gonadotrophin therapy; in the other two, hCG, with or without FSH, caused a rise in inhibin and testosterone, supporting previous observations that LH, as well as FSH, plays a role in the regulation of inhibin or inhibin-related peptides in men.     

丹那唑治疗子宫内膜异位症作用机制的研究

目的:探讨丹那唑治疗子宫内膜异位症(内异症)的作用机制。方法:建立内异症大鼠模型,丹那唑组(n=29)分别皮下注射丹那唑2、4、6、8周,对照组(n=6)皮下注射生理盐水,观察丹那唑对异位内膜生长情况的影响。取丹那唑治疗8周组和对照组的异位内膜,用逆转录聚合酶链反应(RT-PCR)技术,检测丹那唑治疗内异症对异位内膜白细胞介素-6(IL-6)mRNA表达的影响。结果:建模后第3周异位内膜生长良好,各组移植物体积无明显差异(P>0.05)。用丹那唑分别治疗2、4、6和8周后,异位内膜体积缩小(P<0.01),并且用药4、6和8周与用药2周相比,作用更明显(P<0.01)。形态学检查显示,用丹那唑治疗2周后,异位内膜无明显增生;治疗4、6和8周后,异位内膜呈萎缩状态。大鼠成模后给予丹那唑治疗8周,异位内膜IL-6 mRNA的表达明显减少,与对照组相比差异有高度显著性意义(P<0.01)。结论:丹那唑治疗可使内异症大鼠异位内膜逐渐萎缩退化,其作用与用药时间长短有关;丹那唑治疗可使内异症大鼠异位内膜IL-6 mRNA的表达明显降低。     

Antifertility effects of 6-chloro-6-deoxyglucose in the male rat

Male rats were treated with daily oral doses of 12, 24, or 48 mg/kg of 6-chloro-6-deoxy-glucose (6CDG) or with 5 mg/kg of α-chlorohydrin for 28 days. Continuous fertility trials were conducted throughout the dosing period and for 3 weeks following dosing. 12 mg/kg/day of 6CDG produced partial infertility, while 24 and 48 mg produced almost complete infertility after 7 days of dosing. Although the males continued to mate with female rats after this time, only 4 to 20% of mated females became pregnant. In addition, those females which did become pregnant carried a significantly decreased number of fetuses at sacrifice one week after the breeding period. Males dosed with 5 mg/kg/day of α-chlorohydrin showed a gradual decrease in fertility over the four-week dosing period with a significant decrease in fetal numbers observed following one week of dosing. Recovery of normal fertility was achieved after one week withdrawal from all doses of 6CDG or from α-chlorohydrin. The ATP content of spermatozoa obtained from the treated rats and measured following an $\text{in vitro}$ incubation was significantly lower than controls for all groups at 4 weeks of dosing. However, 3 weeks following cessation of treatment, ATP had returned to control levels. The action of 6CDG is rapid in onset and associated with no effect on testis or accessory organ weight. These observations, together with the suppression of intracellular levels of ATP in epididymal spermatozoa, are consistent with an epididymal site of action of this compound or metabolites mediated through the glycolytic pathway.     

Effects of growth factors and hormones on basal and FSH-stimulated inhibin production by porcine granulosa cells in vitro

The effect of several growth factors, protein and steroid hormones on follicle stimulating hormone (FSH)-stimulated and basal inhibin secretion by mature porcine granulosa cells (g-cells) in culture was examined in order to elucidate the putative role of growth factors and hormones in the regulation of inhibin secretion by porcine g-cells in vitro. Cells were incubated with the respective hormones over a timespan of 0-144 h and immunoreactive inhibin was measured with a radioimmunoassay against porcine inhibin. Epidermal growth factor (EGF) and human transforming growth factor type beta (TGF-beta) decreased basal and gonadotrophin-stimulated inhibin and progesterone in a dose-dependent manner. In the absence of insulin, insulin-like growth factor type I (IGF-I) caused a 4-fold enhancement of basal inhibin secretion, but inhibin secretion was elevated only to 20% above control in the presence of 500 nM insulin. Porcine platelet-derived growth factor (PDGF) had no significant effect on basal or FSH-induced inhibin secretion by g-cells. In addition, neither gonadotrophin-releasing hormone (GnRH) nor prolactin (PRL), arginine vasopressin (AVP) and oxytocin affected basal or FSH-stimulated inhibin release by porcine g-cells. Oestradiol caused a slight but significant (P less than 0.01) rise of basal inhibin production (158% of control) in the last 2 days of culture (96-144 h) and the effect of androstenedione on basal (158% of control) and FSH-stimulated (140% of control) inhibin release (P less than 0.01) was also only visible on Days 4-6 of culture. In contrast to androstenedione and oestradiol, progesterone did not show any effect during 6 days of culture in a dose range of 10(-5) to 10(-9) M. Like steroids, prostaglandin E2 (PGE2) had a stimulatory effect on basal inhibin production (250% of control) by porcine g-cells, visible on Days 3-6 of culture, but an inhibitory effect on FSH-stimulated release (less than 40% of control). Over all the experiments with different hormones and growth factors, tested in varying doses and over a time span of 0-144 h, there was a strong correlation between progesterone and inhibin secretion by g-cells (0-48 h = 0.78; 48-96 h = 0.92; 96-144 h = 0.92). These results suggest that EGF, TGF-beta, IGF-I, oestradiol and androstendione as well as PGE2 have para- and/or autocrine modulatory effects on basal and FSH-stimulated inhibin secretion by mature porcine g-cells in vitro and further demonstrate that the secretion of the proteohormone inhibin and the steroid progesterone are closely related.     

HLA-DR和ICAM-1在子宫内膜异位症发病机制中的研究

目的:检测子宫内膜异位症(EM)患者在位和异位子宫内膜中HLA-DR(人类白细胞抗原DR分子)及ICAM-1(细胞粘附分子-1)的表达,探讨它们在EM免疫发病机制中的意义。方法:采用免疫组织化学(SP)法分别测定EM患者在位和异位内膜及正常对照组子宫内膜中HLA-DR和ICAM-1的表达。结果:HLA-DR和ICAM-1在EM患者的异位内膜表达较在位内膜及正常对照组子宫内膜强,差异有统计学意义(P<0.05);HLA-DR和ICAM-1在在位内膜和正常对照组子宫内膜中的表达比较,差异无统计学意义(P>0.05);HLA-DR和ICAM-1在在位子宫内膜的表达呈周期性变化,增生期表达高于分泌期,二者比较差异有统计学意义(P<0.05);正常组子宫内膜的表达在增生期高于分泌期,但差异无统计学意义(P>0.05);HLA-DR和ICAM-1在异位内膜和在位内膜的表达均呈正相关,即同向表达。结论:HLA-DR、ICAM-1在EM患者异位内膜中的表达升高。这些异常变化导致EM患者腹腔局部免疫微环境发生变化,促进了EM的发生发展。     

凋亡调节基因Bcl-2与Bax在子宫内膜异位症中的表达

目的：探讨凋亡调节基因Bcl-2和Bax在子宫内膜异位症发生发展中的作用。方法：SP法检测子宫内膜异位症76例异位内膜、64例在位内膜和72例正常子宫内膜组织中Bcl-2和Bax的表达。结果：异位内膜组Bcl-2蛋白表达强于在位内膜和对照组，分泌期差异有统计学意义（P〈0．05，P〈0．01）；异位内膜组Bax蛋白表达低于在位内膜组和对照组，增生期和分泌期，异位内膜组染色强度均低于在位内膜组和对照组，分泌期差异有统计学意义（P〈0．05）；Bcl-2和Bax在对照组及在位内膜组均呈周期性变化，Bcl-2增生期强于分泌期，Bax分泌期强于增生期，异位内膜组失去周期性变化；增生期3组Bcl-2表达均强于Bax，在位内膜组、异位内膜组差异有统计学意义（P〈0．05，P〈0．01）；而分泌期对照组和在位内膜组Bax强于Bcl-2，异位内膜组Bax表达较Bcl-2弱，差异均有统计学意义（P〈0．05，P〈0．01）。结论：Bcl-2和Bax在子宫内膜异位症发生发展中发挥重要作用。     

Regulation of non-pregnant human uterine contractility. Effect of antihormones

Uterine contractility was recorded on cycle day LH+6 to LH+8 in a control and treatment cycle in 14 healthy non-pregnant volunteers. In the treatment cycle the subjects received either 50 mg of the antiprogestin RU 486 daily for three days or 40 mg of the anti-estrogen tamoxifen daily for two days. The treatment started on day LH+2. During the recording, 2 to 5 micrograms PGF2 alpha was administered into the uterine cavity. The plasma levels of progesterone and estrogen were the same in both the control and treatment cycles. RU 486 caused a significant increase in uterine contractility expressed in Montevideo Units (MU) and a decrease in uterine tonus in comparison with corresponding data obtained in the control cycle. Following treatment with tamoxifen, uterine contractility was lower but the difference was not significant. PGF2 alpha invariably caused a stimulation of uterine contractility. However, treatment with the antihormones did not influence the response. The result of the present study indicates that the change in uterine contractility occurring in the latter part of the menstrual cycle and during menstruation is due to progesterone withdrawal.     

Oral administration of RU 486 and 9-methylene PGE2 for termination of early pregnancy

It has been shown that the antiprogestin RU 486 increases the sensitivity of the early pregnant human uterus to the stimulatory action of prostaglandin E analogues administered vaginally or intramuscularly. To examine if RU 486 also increases uterine sensitivity to a PGE analogue given orally, two investigative approaches were used in the present study: 1) direct registration of uterine contractions before and after administration of 9-methylene PGE₂ in untreated and RU-486-treated early pregnant women; and 2) an efficacy trial involving treatment of pregnant women (amenorrhea of 49 days or less) with 25 mg RU 486 twice daily for three or four days followed by 2.5, 5.0 or 10 mg 9-methylene PGE₂, or 600 mg RU486 followed by 10 mg 9-methylene PGE₂ administered on day 3 and 4. The results showed that oral 9-methylene PGE₂ had a clear stimulatory effect on uterine contractility which was further increased by pretreatment with RU 486. Following 2.5. 5.0 or 10.0 mg 9-methylene PGE₂,the frequency of complete abortion was the same, or approximately 80%. The success rate is higher than that generally reported for RU 486 treatment alone. If 600 mg RU 486 was complemented with 10 mg 9-methylene PGE₂ administered on both days 3 and 4, the frequency of complete abortion increased to 95%. Side effects were of a mild nature and generally occurred following administration of 9-methylene PGE₂. The results of the present study indicate that a combined treatment based on oral administration of both the antiprogestin and the prostaglandin analogue can be developed into a highly effective and simple method to terminate early pregnancy.