Steroidogenesis by Granulosa and Sertoli Cells

Granulosa and Sertoli cells have been isolated from the gonads of immature rats, and have been maintained in monolayer cultures in a chemically-defined medium. The hormonal requirements of these cell types for the synthesis of steroids have been studied: Granulosa cells contain cholesterol side-chain cleavage activity and synthesise progesterone when cultured in the presence of FSH and testosterone (or DHT). Sertoli cells on the other hand cannot synthesise steroids de novo . Granulosa and Sertoli cells are similar in that they are unable to convert significant amounts of progesterone to androgens. In the presence of exogenous testosterone as substrate, granulosa cells and immature Sertoli cells can synthesise estradiol and estrone when stimulated with FSH. In summary, FSH stimulates estrogen synthesis from testosterone in both cell types, and acts synergistically with testosterone (or DHT) to stimulate progesterone synthesis in granulosa cells. FSH is therefore involved in controlling the steroid environment within the gonads.     

Effects of leptin and neuropeptide Y on function of human ovarian granulosa cells in vitro

Objective: To investigate the effects of leptin and neuropeptide Y on steroidogenesis of human ovarian granulosa cells in vitro. Methods: Human ovarian granulosa cells were isolated from follicular fluid obtained during oocyte retrieval of in vitro fertilization-embryo transfer program and cultured for 2 days with various concentration of leptin(1,10,100 ng/ml) or neuropeptide Y (1×10-6, 1×10-7, 1×10-8 mol/L) alone or both,or with the combination of human chorionic gonadotropin (hCG 0, 20 IU/L). The medium was collected for estradiol (E2) and progesterone (P) measurements. Results: (1)Whether hCG existed or not, the adding of leptin did not alter estradiol and progesterone production by human granulosa cells (P>0.05).(2) Only when the concentration of neuropeptide Y was at 1×10-7mol/L,estradiol level was lower than that in the control (P<0.05).(3) The levels of estradiol in neuropeptide Y (1×10-7mol/L) plus hCG group were significantly higher than those with neuropeptide Y alone(P<0.05). (4) In the absence of hCG, the levels of estradiol in neuropeptide Y (1×10-7mol/L)plus leptin (10 ng/ml) group were significantly higher than those with neuropeptide Y(1×10-7mol/L)alone(P<0.05).Conclusions: (1)Leptin alone produced no direct effect on secretion of E2 and P from granulosa cells in vitro.(2)Neuropeptide Y alone may inhibit the secretion of E2, but the inhibition would probably be blocked with the presentation of hCG.(3)Leptin probably blocked the inhibition of neuropeptide Y on E2 secretion, and this may indicate that there were some coordination between leptin and neuropeptid Y on the level of ovarian function.     

瘦素、神经肽Y对离体人卵巢颗粒细胞分泌功能的影响

目的　探讨瘦素、神经肽Y(NPY)对离体人卵巢颗粒细胞雌、孕激素生成的影响。　方法　将来自体外受精 胚胎移植的 2 5例离体人卵巢颗粒细胞纯化后 ,在不同浓度瘦素 (1、10、10 0 μg/L)、NPY(1× 10 -8、1× 10 -7、1× 10 -6mol/L)单独或与人绒毛膜促性腺激素 (hCG ,0、2 0IU/L)联合作用下体外培养 ,采用放射免疫方法测定培养液中雌二醇 (E2 )、孕酮(P)的水平。　结果　 (1)无论单独或与hCG联合不同浓度瘦素组E2 、P水平均无显著性差异 (P >0 .0 5 ) ;(2 )NPY浓度 1× 10 -7mol/L时 ,E2 水平显著低于空白对照组 (P <0 .0 5 ) ,但与hCG联合组显著高于单独组 (P <0 .0 5 ) ;(3)NPY浓度 1× 10 -7mol/L、瘦素浓度 10 μg/L组 ,E2 水平明显高于相应浓度NPY单独作用组 (P <0 .0 5 )。　结论　 (1)瘦素不直接影响体外培养颗粒细胞E2 、P的分泌 ,可能通过影响其它因子的作用而间接调节卵巢功能 ;(2 )NPY单独作用时 ,对颗粒细胞E2 的分泌有一定抑制作用 ,但当hCG存在时作用消失 ,说明在正常月经周期中NPY对黄体的生成和维持可能无明显不利影响 ;(3)瘦素可能阻断NPY对颗粒细胞分泌E2 的抑制作用 ,表明二者可能对卵巢功能有相互协调作用。     

转铁蛋白抑制FSH刺激大鼠卵泡颗粒细胞分化的细胞内主要机制

前期的研究已经证实了转铁蛋白(TRF)能抑制FSH诱导的抑制素、雌二醇和孕酮的产生以及芳香化酶活性。本实验用已烯雌酚处理后分离的大鼠卵泡颗粒细胞的体外培养方法,进一步观察TRF对FSH、霍乱毒素(cholera toxin,CT)及Forskolin诱导的cAMP、雌二醇及孕酮产生的影响。结果:TRF抑制了FSH诱导的cAMP、雌二醇及孕酮产生TRF能抑制CT及Forskolin诱导的雌二醇产生,但不影响这些因子刺激cAMP和孕酮生成。说明TRF抑制FSH对大鼠卵泡颗粒细胞功能性分化刺激作用的主要细胞内机制是通过减少cAMP生成而介导的,但还有cAMP以外的作用部位。     

细胞外基质对大鼠卵巢颗粒细胞分泌激素和合成蛋白质功能的影响

为了探讨细胞外基质对卵巢颗粒细胞功能的影响。从未成熟ＳＤ大鼠分离出卵巢颗粒细胞，以３×１０５细胞／孔的密度，培养在铺有不同浓度的层粘连蛋白（ＬＮ）或纤粘连蛋白（ＦＮ）的培养板上，观察细胞贴壁情况。４８小时后加入卵泡刺激素和睾酮，再培养４８小时，收集培养液，测定雌二醇和孕酮。另外在双池培养系统中，在铺有和未铺ｍａｔｒｉｇｅｌ的上池中分别加入不同浓度的颗粒细胞，利用３５硫标记蛋氨酸掺入法观察ｍａｔｒｉｇｅｌ对颗粒细胞合成蛋白的影响，并利用免疫印迹法鉴定颗粒细胞膜蛋白中ＬＮ结合蛋白的分子量。结果显示，４μｇＬＮ即可使颗粒细胞在４小时内贴壁及展开，雌二醇和孕酮的产量增加。ＦＮ却无显著影响。从ｍａｔｒｉｇｅｌ双池培养系统中取上池培养液，测定放射活性，铺有ｍａｔｒｉｇｅｌ组显著高于对照组。放射自显影图象显示ｍａｔｒｉｇｅｌ显著刺激颗粒细胞合成功能。免疫印迹结果显示颗粒细胞膜蛋白存在着ＬＮ的多结合位点。本实验证实：ＬＮ对卵巢颗粒细胞的附着、伸展、分泌和合成功能具有促进作用     

Interleukin-4 and interleukin-13 potentiate interleukin-1 induced secretion of interleukin-6 in human osteoblast-like cells.

Formation of interleukin-6 (IL-6) in osteoblasts and bone marrow stromal cells is believed to regulate osteoclast recruitment. The anti-inflammatory cytokines interleukin-4 and -13 (IL-4 and IL-13) stimulate IL-6 production in human osteoblasts. We investigated the relative potencies, and synergistic effects, between IL-4, IL-13 and interleukin-1 (IL-1) on IL-6 formation in human osteoblast-like cells. Isolated human osteoblast-like cells were incubated for 72 h in the presence of various concentrations of IL-4, IL-13 and IL-1, and IL-6 secretion was measured by ELISA. All cytokines stimulated the secretion of IL-6. The rank order of potency was IL-1>IL-4>IL-13. There were no additive or synergistic effects between IL-4 and IL-13. However, co-stimulation with IL-1 and IL-4 resulted in a marked synergistic effect on IL-6 secretion. Co- stimulation with IL-1 and IL-13 gave a minor synergistic effect. In conclusion, IL-4/13 synergistically potentiates IL-1 induced secretion of IL-6 in human osteoblast-like cells.     

溴氰菊酯对大鼠卵巢甾体激素合成的影响

目的探讨溴氰菊酯(Deltamethrin,DLT)对大鼠卵巢甾体激素合成的影响及相关的分子机制。方法不同剂量DLT暴露处理大鼠和原代颗粒细胞;取DLT暴露后的大鼠血清和培养上清液,电化学发光法测定雌二醇和孕酮的水平;Western Blot法检测DLT暴露后大鼠卵巢组织及原代培养颗粒细胞中甾体激素合成关键酶:甾体激素合成急性调节蛋白(StAR)、胆固醇侧链裂解酶(P450scc)、芳香化酶的表达量,同时检测组蛋白去乙酰化酶Sirt1的蛋白表达。结果体、内外实验中,DLT各处理组大鼠血清、原代颗粒细胞培养上清液中雌二醇水平均显著升高(体内实验P<0.05,体外实验P<0.001);体内实验中DLT各处理组大鼠血清孕酮水平均显著降低(P<0.05),而体外实验中DLT各处理组原代颗粒细胞培养上清液孕酮水平无显著变化(P>0.05);体、内外实验中DLT各处理组大鼠卵巢组织、原代培养颗粒细胞中StAR、P450scc、芳香化酶以及Sirt1的表达量均显著升高(P<0.01)。结论 DLT能影响大鼠卵巢雌二醇和孕酮的合成,其机制可能与其关键酶表达的变化有关。     

Specific regulatory actions of dihydrotestosterone and estradiol on the dynamics of FSH secretion and clearance in humans

The authors investigated immunoactive and bioactive follicle-stimulating hormone (FSH) secretion and clearance in six healthy young men during steady-state infusions of vehicle (basal, B, 28 hours), dihydrotestosterone (DHT, 4.5 days), or estradiol (4.5 days) accompanied by blood sampling at 10-minute intervals for 28 hours. Serum FSH concentrations were assayed by a two-site immunoradiometric assay (IRMA) and two separate in vitro bioassays (rat granulosa and Sertoli cell systems). FSH measurements included: 24-hour mean serum concentrations (IRMA and bioassay), multiple-parameter deconvolution of 24-hour pulsatile FSH time series and FSH release in response to exogenous gonadotropin-releasing hormone (GnRH) boluses (IRMA) to assess secretion and clearance, and circadian serum FSH concentration rhythms by cosinor analysis (IRMA). We found: 1) a significant decrease in 24-hour mean IRMA FSH concentrations during DHT infusion while both in vitro estimates of FSH bioactivity were unchanged; 2) significant decreases in the mass of IRMA FSH secreted per 24 hours during DHT infusion; 3) significant decreases in the IRMA FSH half-life during estradiol infusion without any change in FSH interpulse interval; 4) no steroidal effects on FSH secretory responses to exogenous GnRH; and 5) abolition of basal circadian FSH rhythms during sex-steroid infusions. Based on these findings, we conclude that steady-state sex-steroid hormone infusions selectively alter IRMA FSH secretion and clearance without affecting IRMA FSH pulse frequency or mean concentrations of bioactive FSH.     

GnRH拮抗剂联合来曲唑和米非司酮对体外培养的人颗粒细胞功能的影响

目的探讨促性腺激素释放激素拮抗剂(GnRH-ant)联合来曲唑和米非司酮对体外培养的人卵巢颗粒细胞功能及血管内皮生长因子(VEGF)表达的影响。方法体外培养人原代卵巢颗粒细胞及人卵巢颗粒细胞系KGN,根据所用药物不同分为GnRH-ant、来曲唑、米非司酮以及上述三种药物联合用药组,药物干预后检测细胞活力、细胞凋亡、培养液中雌二醇(E 2)及孕酮(P)水平及VEGF的表达。结果(1)GnRH-ant、来曲唑、米非司酮三种药物均可以不同程度地抑制原代颗粒细胞和KGN细胞的活力、促进颗粒细胞凋亡,其中联合用药组的作用最强(P<0.05);(2)GnRH-ant、来曲唑、米非司酮三种药物均可以不同程度地抑制原代颗粒细胞和KGN细胞E 2、P分泌水平,其中联合用药组E 2、P水平最低且差异有统计学意义(P<0.05);(3)GnRH-ant、来曲唑、米非司酮三种药物均可以不同程度地抑制原代颗粒细胞和KGN细胞VEGF表达,其中联合用药组VEGF mRNA及蛋白表达水平最低(P<0.05)。结论GnRH-ant联合来曲唑和米非司酮可以有效地抑制颗粒细胞活力、减少类固醇激素分泌及降低VEGF的表达水平。     

c-ras与大鼠颗粒细胞和黄体细胞生成孕酮的关系

应用离体细胞培养 ,研究了反义 c-ras寡脱氧核苷酸 (反义 c-ras ODN)对人绒毛膜促性腺激素 (h CG)诱导的大鼠颗粒细胞和黄体细胞生成孕酮 (P)的影响及其与外源性 c AMP和 Ca2 +的关系。结果发现 ,2 0 μmol/ L 反义 c-ras ODN能显著抑制 h CG诱导颗粒细胞的 P生成量 (P<0 .0 5 ) ,而对黄体细胞的 P生成无明显影响 ;反义 c-ras ODN对h CG诱导的颗粒细胞生成 P的抑制作用能被加入 1 0 - 4mol/ L 二丁酰 c AMP所逆转 ,钙通道阻断剂维拉帕米对抑制作用具有协同效应。结果提示 ,c-ras癌基因参与 h CG诱导的颗粒细胞生成 P的调控 ,而对 h CG诱导的黄体细胞 P生成关系不大     

流式细胞计数测定抗孕激素对猪卵巢细胞孕酮受体的影响

为了研究抗孕激素调节卵巢功能的机理 ,制备了人胎盘羊膜的双池培养系统。分离猪卵巢颗粒细胞和内泡膜细胞 ,分别种在羊膜两侧。在加入或不加 RU486和ZK98.73 4时 ,两种细胞在促性腺激素刺激下共同培养。 48h后 ,应用流式细胞计数(FCM)测定两种细胞的孕酮受体 (PR)和雌激素受体 (ER) ;放射免疫法测定培养液中的孕酮 (P)和雌二醇 (E2 )浓度。结果显示 ,抗孕激素增加两种细胞的 PR含量 ,ER没有明显改变 ;颗粒细胞的 P和 E2 分泌受到抑制 ,内泡膜细胞的 P分泌也减少。提示 P调节 P本身的合成 ,这种旁分泌和自分泌作用是通过 P控制 PR上调的途径。这可能是抗孕激素对卵泡发育和甾体激素生成功能的抑制机理之一     

Effects of progesterone administration on follicle-stimulating hormone and prolactin release in estrogen treated eugonadal adult men

In order to investigate the modulatory effect of steroids on FSH secretion in vivo, we studied 16 human males, aged 51-81 years, affected by prostatic carcinoma. They were given estradiol or E2 plus progesterone (P), added at different times during E2 treatment. Daily blood samples were collected in order to determine LH, FSH, and PRL levels; moreover, blood samples were collected at 2 h intervals for 12 h on the day of P administration. We observed the expected biphasic effect on LH secretion, whereas daily basal FSH levels, during E2 treatment, decreased gradually and progressively from the first day until the end of the study. FSH levels exhibited, after P administration, wide fluctuations, with peak levels observed from 2 to 6 h after P in 4 of 6 patients studied (at 72 h during E2 treatment). A clear trend toward FSH increase was also observed in 3 out of 5 patients in whom P was administrated 96 h after starting E2 administration. In this case, FSH increases were delayed, becoming evident between 8th and 10th h after P injection. Finally, during E2 administration basal PRL levels showed a progressive increase, which was significant in all three groups. In conclusion, these data confirm the biphasic effects of estrogen administration on LH secretion in eugonadal adult human males; while estrogens alone showed an inhibitory effect on FSH secretion, the addition of P induced also a positive action, resulting in a clear FSH peak in some patients tested. The time course of E2 and P administration seems to be critical for the hormone response pattern.(ABSTRACT TRUNCATED AT 250 WORDS)     

Interaction between porcine granulosa and thecal cells in steroidogenesis in an amnion dual chamber culture system

IntroductionInordertoinvestigatetheovariansteroidogenicfunction,scientistsusedtocul-turegranulosacellsorthecalcellsinvit-roLl~5].Thereisagrowingawarenessthatinvivogranulosaandthecalcellsintheovaryareseparatedjustbyabasementmetnbrane.Bothtypesofcellsareimportantduringthefollicledevelopmentandovummaturation,interact-ingoneachother.The"twocells"theoryhasbeenknownformorethantwentyyears['].Thesinglecellculturewitheithergranulosacellsorthecalcellswasnotaperfectmodelforinvitrostudy,Itisnecessarytoe…     

Does isoflurane enhance rhinovirus replication and virus‐induced cytokine secretion in airway epithelial cells?

Background: There have been no previous studies regarding the effect of volatile anesthetics on human rhinovirus (RV) infection in airway epithelial cells of patients with an upper respiratory infection (URI). We have therefore evaluated in vitro the effect of isoflurane on RV infection in airway epithelial cells.
Methods: A549 cells and RV-infected A549 cells were treated with isoflurane for 2 or 4 h. Surface expression of intercellular adhesion molecule-1 (ICAM-1) was assessed by flow cytometry, and effects on the secretion of interleukin (IL)-6 and IL-8 were measured by ELISA. The effect on RV replication in the cells was determined by viral titer.
Results: Isoflurane treatment for 2 or 4 h had no significant effect on ICAM-1 expression and secretion of IL-6 and IL-8 in control cells. Isoflurane also had no significant additional effect on RV-induced ICAM-1 expression and secretion of IL-6 and IL-8. Viral titers were not significantly influenced by isoflurane.
Conclusion: Isoflurane treatment showed no additional effects with RV on ICAM-1 expression, secretion of IL-6 and IL-8, and viral titer in A549 cells. These results suggest that isoflurane itself may not increase further RV infections, inflammations, and viral replication in patients with a viral URI.     

Effect of Bisphenol A on steroid production in human granulosa cells in vitro

Objective:To study the effects of environmental estrogen-like chemical bisphenol A(BPA)on ovarian function with the model of the cultured human granulosa cells in vitro.Methods:The granulosa cells from healthy women were collected and cultured in DMEM with 10% heat-inacti-vated fetal bovine serum.BPA,final concentration 10-7 to 10-4 mol/L,was added to the cell-culture medium and treat cells for 48 hours.The levels of estrogen and progesterone in the supernatant of the cultured cells were meas-ured by DELFIA.Total RNA was extracted from the cultured cells.The expression levels of P450 scc and P450 arom mRNA were measured by RT-PCR.Results:The cultured human granulosa cells could express high levels of P450scc and P450arom.The levels of estrogen and progesterone increased after BPA treatment,expression of P450 arom mRNA was reduced significantly at 10-5 to 10-4mol/L of BPA,but the expression of P450 scc mRNA was increased at 10-6 to 10-5 mol/L of BPA.The levels of estrogen and progesterone in the supernatant of the cultured cells were not affected significantly by BPA at the same concentrations incubated for 48 hours.Conclusion:BPA could affect steroid hormone synthesis and transformation in granulosa cells,such as the ex-pression of P450 scc and P450 atom,so it can affect ovarian function.Although we did not find a significant effect of BPA on the final estrogen and progesterone levels in this studying model,noxious effects of BPA on ovarian function may be exist in the human granulose cells.     

酪氨酸、米非司酮和睾酮对大鼠颗粒细胞凋亡的影响

大多数哺乳类动物的卵巢卵泡在排卵前闭锁，研究表明其发生机制是细胞凋亡。本实验观察了卵巢调控因子酪氨酸、米非司酮和睾酮对大鼠卵巢颗粒细胞凋亡的影响。结果：米非司酮和睾酮促进凋亡，造成ＤＮＡ的片段化反应；而酪氨酸的作用则相反。与对照组相比，米非司酮和睾酮组血清雌二醇含量下降而孕酮含量升高；酪氨酸组血清雌二醇水平升高而孕酮水平下降。米非司酮和睾酮增加闭锁卵泡数，酪氨酸作用则相反。     

Impaired release of interleukin-6 from human osteoblastic cells in the uraemic milieu.

BACKGROUND: Osteoblast-derived interleukin-6 (IL-6) affects bone metabolism and is linked with a number of pathological states characterized by increased bone resorption, including osteoporosis and renal osteodystrophy. To examine the possibility that uraemia directly influences the release of this cytokine in bone, we have investigated the effect of human uraemic serum on the release of IL-6 from human osteoblast-like cells. METHODS: Individual serum samples collected from healthy male volunteers or male haemodialysis patients prior to and during a dialysis treatment were assayed for IL-6, interleukin-1beta (IL-1beta) and soluble IL-6 receptor (sIL-6R) using specific enzyme-linked immunosorbent assays. MG-63 and SaOS-2 cells were cultured in media containing pooled sera from both groups and alongside matching charcoal-stripped sera. IL-6 concentrations were determined in harvested cell supernatants after 24 h. In further experiments, media containing individual sera obtained from five patients at regular intervals during their haemodialysis treatment were incubated with MG-63 cells to determine the effects of the dialysis process on IL-6 secretion. RESULTS: Haemodialysis patients had significantly higher (n = 10, P < 0.001) circulating concentrations of IL-6 (7.0 +/- 1.6 pg/ml) than normal subjects (0.4 +/- 0.1 pg/ml), but there were no significant differences in the concentrations of either IL-1beta or sIL-6R. These serum concentrations did not change significantly during 80 min of dialysis. IL-6 release by MG-63 cells incubated with charcoal-stripped serum from normal or from uraemic subjects was similar. Incubation with untreated sera from normal subjects increased IL-6 release by approximately 6-fold above the charcoal-stripped control, whereas sera from uraemic subjects increased IL-6 release by only approximately 2- to 3-fold (normal vs uraemic of 6878 +/- 595 and 2579 +/- 169 pg/ml, respectively, P < 0.001). Similar results were seen with SaOS-2 cells. Haemodialysis did not restore the capacity of uraemic serum to augment IL-6 release to the same degree as normal serum. CONCLUSIONS: These data show that the augmentation of IL-6 release from human osteoblastic cells after incubation with normal serum is greater than after uraemic serum. This may indicate the presence of an inhibitor of IL-6 release in uraemic serum that is involved in the deranged bone turnover of uraemic patients.     

NIMA相关蛋白激酶7、Nod样受体蛋白3与多囊卵巢综合征炎症指标相关性研究

目的探讨卵巢颗粒细胞中NIMA相关蛋白激酶7(NEK7)和Nod样受体蛋白3(NLRP3) mRNA表达量与多囊卵巢综合征(PCOS)患者炎症指标的相关性及其预测价值。方法选取2018年8月至2019年3月来山西省儿童医院/山西省妇幼保健院生殖中心就诊的30例PCOS患者为研究组和同期由于男方因素或输卵管因素就诊的34例非PCOS不孕症患者为对照组。取卵日收集行IVF/ICSI-ET患者的卵泡液和颗粒细胞,检测卵泡液中炎症因子白介素(IL)-1β、IL-18的表达以及颗粒细胞中NEK7、NLRP3 mRNA的表达,并将NEK7、NLRP3的mRNA表达量与各指标进行相关性分析。最后应用受试者工作特征曲线(ROC)分析NEK7、NLRP3 mRNA对PCOS的预测价值。结果研究组体重指数(BMI)和获卵数显著高于对照组,且LH、LH/FSH、睾酮(T)水平亦显著高于对照组(P均<0.05)。研究组卵泡液中炎症因子IL-1β、IL-18的表达显著高于对照组,卵巢颗粒细胞中NEK7、NLRP3 mRNA的表达亦显著高于对照组(P均<0.05)。相关性分析结果显示,研究组患者卵巢颗粒细胞中NEK7和NLRP3 mRNA水平与血清基础LH、LH/FSH、炎症因子IL-1β、IL-18均呈显著正相关(P均<0.05)。ROC分析结果显示,NEK7是PCOS的有效预测因子,预测炎症的曲线下面积为0.741,临界值为1.154。结论 NEK7、NLRP3可能通过促进IL-1β、IL-18的释放影响PCOS炎症的发生及发展,且NEK7是PCOS的有效预测因子,可能为PCOS的临床治疗提供新思路。     

Observation on human ovarian theca cell and granulosa cell interaction in polycystic ovary syndrome

<正> Objective:To investigate the role of human theca eell(TC) and granulosa cell (GC)interaction and insulin(INS) in steroidogenesis in normal ovarian cycle and in patientswith PCOS.Methods:Ovarian theca and granulosa cells from eleven normal women and eightPCOS patients were co-cultured on opposite side of collagen with or without INS.Theconcentrations of estradiol (E_2),progesterone (P) and androstenedione (A) in the cul-ture medium were examined by ELISA method.Results:When co-cultured with GC,TC in PCOS group produced more A and lessP than those of normal group.When co-cultured with theca cells,granulosa cells inPCOS group produced more E_2 than those of normal group.Addition of INS increasedthe difference significantly.Conclusions:The GC and TC interaction from the normal and PCOS ovaries is dif-ferent.There is a high A and high E_2 intraovary loop of PCOS leading to premature ar-rest of follicle growth and anovulation.Insulin may play an important regulatory role.     

卵泡基膜中IV型胶原、层粘连蛋白来源的初步探讨

通过离体培养猪、大鼠卵泡颗粒细胞、内泡膜细胞，应用免疫组化技术，观察细胞外基质与卵泡细胞结构和功能的关系，初步探讨卵泡基膜中ＩＶ型胶原、层粘连蛋白（ＬＮ）的主要来源及其生物学意义。结果表明：内泡膜细胞合成、分泌大量的ＩＶ型胶原，颗粒细胞不合成ＩＶ型胶原；颗粒细胞和泡膜细胞合成、分泌大量ＬＮ。由于ＩＶ型胶原属于不溶性蛋白，ＬＮ属于微溶性蛋白，因此卵泡基膜中的ＬＮ主要来源于基膜两侧的颗粒细胞、内泡膜细胞，ＩＶ型胶原主要来源于内泡膜细胞。体外无血清培养大鼠颗粒细胞，单独铺有细胞外基质的主要成分纤粘连蛋白（ＦＮ）、ＬＮ、ＩＶ型胶原时，明显促进颗粒细胞的增殖、分化，孕酮分泌增加；在同时铺有这三个细胞外基质成分时，上述作用最强，颗粒细胞分泌ｃＡＭＰ明显增加，说明ＦＮ、ＬＮ、ＩＶ型胶原具有协同作用，同时说明它们促进颗粒细胞分泌孕酮作用，是通过刺激ｃＡＭＰ水平升高实现的。