ANGIOTENSIN II RECEPTOR DENSITY IN BOVINE OVARIAN FOLLICLES RELATES TO TISSUE RENIN AND FOLLICULAR SIZE

1. The purpose of this study was to investigate angiotensin II (AII) receptors in isolated bovine ovarian follicles and the relationship of their density to follicular concentrations of prorenin, active renin, oestradiol and progesterone. 2. Displacement of [¹²⁵I]-[Sar¹-Ile⁵-Ile⁸]-AII binding by the AII receptor antagonists PD 123319 and Losartan (DuP 753) confirmed that follicular AII receptors are of subtype 2 (AT₂ receptor). 3. The dissociation constant (K_d) for [Ile⁵]-AII (human AII) was 0.84 (range 0.51–1.47) nmol/ L. The receptor density varied between 90 and 5990 (mean 1640) fmol/ mg membrane protein. 4. The follicular AII receptor density correlated positively with follicular diameter (Spearman's rho= 0.518; P<0.003) and tissue weight (Spearman's rho= 0.636; P<0.0001), and negatively with the active renin concentration in the follicular wall (Spearman's rho=-0.399; P<0.02). The AII receptor density did not correlate with the follicular fluid concentrations of prorenin, active renin, oestradiol (E₂, progesterone (P₄) or the E₂/P₄ ratio. The follicular fluid concentrations of prorenin correlated negatively with the E₂/P₄ ratio (Spearman's rho= -0.716; P<0.00001). 5. The inverse relationship between AII receptor density and the high active renin concentrations in the follicular wall suggests an active regulated tissue renin-angiotensin system. A high AII receptor density is a general feature of large bovine ovarian follicles.     

MEASUREMENT AND IDENTIFICATION OF PRORENIN AND RENIN IN OVARIAN FOLLICULAR FLUID FROM CATTLE AND PIG

1. In previous studies we have demonstrated and solved several methodological problems in relation to the measurement of prorenin by trypsin activation in rat, bovine, hog and horse plasma. 2. The aim of the present study was to develop a method for the measurement of prorenin in bovine and porcine ovarian follicular fluid. 3. Trypsin activation of follicular fluid generated angiotensin I immunoreactive material (AI IM) in both species. 4. The AI IM interfered with the renin assay, but could be completely removed by a cation exchange resin in a batch-wise technique. 5. The enzymatic activity of trypsin-activated prorenin and pre-existing active renin was completely inhibited by a specific inhibitor of renin. 6. The reactions were optimized and an accurate measurement of prorenin in ovarian follicular fluid was developed. 7. The existence of prorenin and renin in bovine ovarian follicular fluid was established. Prorenin and renin in porcine ovarian follicular fluid was demonstrated for the first time. 8. The ratio between ovarian follicular fluid and plasma was 43 for prorenin and 19 for active renin in cattle. The same ratios in pigs were 1.3 and 0.4, respectively. These findings indicate a species difference with respect to the amount of prorenin or active renin present in ovarian follicular fluid.     

IN VITRO INCUBATION OF BOVINE OVARIAN FOLLICLES: INDICATIONS FOR AN ACTIVE AND REGULATED RENIN-ANGIOTENSIN SYSTEM

1. The ovarian follicular renin—angiotensin system was investigated by using an in vitro incubation method, based on tissue incubation of individual bovine follicles. 2. Very high and varying concentrations of active renin (median 18.0 GU/kg, range 2.1–107 GU/kg; n= 101) and prorenin (11.7 GU/kg, range <2.6–142 GU/kg; n = 101) existed in unincubated ovarian follicular tissue. 3. Active renin and prorenin increased 35 and 959%, respectively, in follicular wall tissue and incubation medium during 72 h of in vitro incubation. The protein synthesis inhibitor cyclohexamide inhibited active renin and prorenin formation. No activation of prorenin occurred in incubation medium or follicular fluid in vitro. Active renin was degraded during incubation. 4. Primarily prorenin but also active renin were secreted into the incubation medium. Secretion was directed both to the internal and external surface of the follicular wall. 5. Follicle-stimulating hormone (FSH) slightly stimulated the synthesis and secretion of prorenin in vitro. 6. The synthesis and secretion rates of active renin and prorenin varied markedly between individual follicles. 7. The finding of very high and varying concentrations of active renin in follicular wall tissues together with the formation and secretion of active renin during in vitro incubation provide further evidence for an active and regulated renin-angiotensin system in ovarian follicles.     

RENIN AND PRORENIN IN REPRODUCTIVE TISSUES DURING GESTATION IN PIGS AND CATTLE

1. High concentrations of prorenin and active renin were previously found in ovarian follicular fluid from cattle but not from pigs. In the present study female reproductive tissues and fluids from cattle and pigs during gestation were investigated to clarify a possible species difference in active renin and prorenin concentrations. 2. Very high concentrations of active renin but no prorenin were found in corpus luteum from both species. 3. Relatively low concentrations of active renin, in the same order as in maternal blood plasma, were found in myometrium, endometrium, placenta and fetal membranes from both species. Prorenin was undetectable in these tissues except for bovine myometrium and porcine endometrium in some animals. 4. The concentrations of active renin and prorenin in amnionic fluid from both species were below the maternal plasma values. In allantoic fluid the concentrations were higher than in amnionic fluid. 5. The plasma concentrations of active renin and prorenin did not change during gestation in pigs. This finding is in contrast to the observations in humans and does not support a systemic effect of prorenin during gestation. 6. The presence of renin in the reproductive tissues, especially the very high concentrations in the corpus luteum, indicates a local function of the renin-angiotensin system during gestation.     

CIRCULATING LEVELS OF ACTIVE, TOTAL AND INACTIVE RENIN (PRORENIN), ANGIOTENSIN I AND ANGIOTENSINOGEN IN CARBON TETRACHLORIDE-TREATED RATS

1. Plasma renin activity (PRA), plasma angiotensin I concentration (ANG I), plasma angiotensinogen concentration (PAC) and the plasma levels of active, total and inactive renin (prorenin) were measured in rats with carbon tetrachloride (CCl₄)-induced acute renal failure. Rats were treated with a single oral dose of CCl₄ (2.5 mL/kg) and killed 1, 2, 3 and 7 days later. 2. On days 1–3 PRA, ANG I and PAC decreased and increased on day 7. Active renin fell on days 2 and 3, total renin (trypsin treatment) augmented on day 1 and diminished on day 3, prorenin and per cent prorenin increased on days 1 and 2. Angiotensin I concentration paralleled PRA and PAC. The CCl₄-induced decrease in PRA was secondary to the fall in active renin and in PAC. Total renin augmented as a consequence of the elevation of prorenin. Renal function, evaluated by serum urea, serum creatinine and creatinine clearance, decreased on days 1 and 2 when PRA was low and plasma prorenin was high. 3. These data do not support the involvement of the circulating active renin-angiotensin system (RAS) in the pathophysiology of acute renal failure induced by CCl₄, however, increased prorenin levels were associated with the decrease in renal function.     

EFFECT OF GONADOTROPINS AND PREGNANCY ON PRORENIN AND RENIN IN THE RAT

1. Stimulation of adult female rats with pregnant mare serum gonadotropin (PMSG) and human chorion gonadotropin (hCG) increased active plasma renin about two-fold, but caused only a slight increase of plasma prorenin. The concentrations of active renin and prorenin in the ovaries, and active renin in the uterus all increased about two-fold 2 days after stimulation with PMSG. The prorenin in the uterus was below detection in unstimulated rats and did not change consistently after PMSG. 2. Active renin and prorenin in plasma were unchanged in relation to pregnancy, except for a slight decrease of prorenin in the third trimester. In the first and third trimester the concentration in the ovaries of active renin and prorenin was decreased to about one-third of that in normal female rats. In contrast active renin in the uterus was increased about two-fold in the first trimester, whereas prorenin did not change consistently. 3. Our results confirm that gonadotropins and pregnancy affect the renin-angiotensin system in rats. However, the changes in the plasma seem to be much smaller than those previously reported in humans. Accordingly, our results do not support a systemic role of prorenin for reproduction in the rat.     

Effects of exogenous female sex-steroid hormones on lymphocyte β2-adrenoceptors in normal females

We have previously shown that lymphocyte β₂-adrenoceptors (AR) are under cyclical control of sex-steroid hormones with greater receptor density during the luteal phase of the menstrual cycle. It has also been postulated that abnormal cyclical regulation of β₂-AR might be a possible mechanism for premenstrual asthma. The effects of exogenous female sex-steroid hormones on lymphocyte β₂-AR function were studied in eight normal healthy females. They were evaluated at two successive menstrual cycles, during the follicular phase (day 1–6). They were randomized to receive single oral doses of either ethinyloestradiol 50 μg or medroxyprogesterone 10 mg in a cross-over study. Lymphocyte β₂-AR parameters were evaluated at baseline (t₀), 24 h (t₂₄) and 72 h (t₇₂) after ingestion. Baseline levels of progesterone and oestradiol were comparable on both cycles. Receptor density (B_max) increased significantly (P<0.01) from t₀ after progesterone but not oestradiol at t₂₄: a 1.39-fold geometric mean difference (95% CI 0.96–2.00) between t₂₄vs t₀. Receptor affinity (K_d) and maximal cAMP response to isoprenaline (E_max) were not altered by either treatment. These results show that exogenous progesterone but not oestradiol, given during the follicular phase, significantly increased β₂-AR. This, therefore, suggests that endogenous progesterone is probably responsible for previously observed increase in B_max during the luteal phase of the female menstrual cycle. These findings may suggest possible therapeutic strategies for modulation of β₂-AR in premenstrual asthma.     

肾素前体受体依赖性诱导大鼠血管平滑肌细胞增殖和肥大

目的研究肾素前体通过非血管紧张素Ⅱ途径诱导大鼠血管平滑肌细胞的增殖和肥大。方法利用3 H同位素标记的胸苷和亮氨酸的结合检测血管平滑肌细胞增殖和肥大。肾素受体siRNA进行受体表达抑制。结果肾素前体处理细胞48h后剂量依赖性诱导细胞内3 H同位素标记的胸苷和亮氨酸含量增高(n=6,P<0.05)。而且,20nmol/L肾素前体处理24h后细胞内3 H同位素标记的亮氨酸显著增高(n=6,P<0.05)。预处理血管紧张素Ⅱ受体抑制剂(ARB)坎地沙坦(100nmol/L)和血管紧张素转换酶(ACE)抑制剂卡托普利(1mmol/L)对20nmol/L肾素前体诱导的细胞增殖和肥大没有作用,然而对于肾素受体siRNA处理后的细胞肾素前体不能引起细胞的增殖和肥大。结论肾素前体诱导血管平滑肌细胞产生的增殖和肥大效应是肾素受体依赖的,而不是通过血管紧张素Ⅱ的产生发挥作用。     

Growth hormone responses to GABAB receptor stimulation throughout the menstrual cycle of healthy females

Growth hormone (GH) responses to the GABA_B agonist baclofen (10 mg) were assessed in six normally cycling, healthy women at three different sex steroid phases of the menstrual cycle: early follicular, pre-ovulatory and luteal. The design was placebo-controlled, balanced and single-blind with a total of six tests being carried out on each subject in two consecutive menstrual cycles. GH responses to baclofen increased incrementally from early-through mid- to late-cycle: 10.95 ± 0.95 μg/l, 24.53 ± 4.73 μg/l, 31.36 ± 3.37 μg/l respectively (p < 0.04). Responses between early- and mid-cycle and early- and late-cycle were significantly different (p < 0.05). There was a direct relationship between baclofen/GH responses and both plasma oestradiol (E₂) concentrations (p = 0.05) and plasma progesterone concentrations (p = 0.02). Responses to placebo did not vary. E₂ has been demonstrated to exert a priming influence on GH responses to many pharmacological and physiological stimuli. An E₂-induced priming effect on GH secretion may underlie these results. Progesterone may augment this effect. Other possible influences are a sex steroid effect on somatostatin or GABA neurotransmission.     

AUTORADIOGRAPHIC LOCALIZATION OF ACTIVE RENIN IN THE JUXTAGLOMERULAR APPARATUS OF THE DOG KIDNEY: EFFECTS OF SODIUM INTAKE

1. The effects of dietary sodium intake on active renin binding in the juxtaglomerular apparatus (JGA) of superficial and juxtamedullary cortex of the dog kidney were examined by quantitative in vitro autoradiography using a radiolabelled renin inhibitor [¹²⁵I]-H77, which has high affinity for dog renin. 2. Changes in sodium intake resulted in marked alterations of active renin binding in the radiolabelled JGA. In comparison with the control kidney (190.8 ± 7.7 Bq/mm³), a higher density of binding occurred in the labelled JGA of sodiumdepleted kidney (277.7 ± 6.2 Bq/mm³), while a lower density of binding was found in the labelled JGA of sodium-loaded kidney (99.3 ± 7.4 Bq/mm³). 3. Active renin binding in the labelled JGA was significantly higher in superficial JGA than in their juxtamedullary counterparts, irrespective of sodium intake. 4. Pre-incubation with trypsin (0.5mg/mL), a procedure known to activate prorenin, markedly increased active renin binding in the labelled JGA of control (+～ 35%; P < 0.01), and sodium-loaded kidneys (+～ 75%; P < 0.01), but had little effect on binding in the labelled JGA of the sodium-depleted kidney (± 5–10%; NS). The proportions of active renin as a percentage of total renin were 60, 75 and 95% in the labelled JGA of sodium-loaded, control, and sodium-depleted kidneys, respectively. 5. Emulsion microscopic autoradiography revealed that the binding was exclusively localized in the JGA, including the afferent and efferent arterioles, macula densa and extraglomerular mesangium. Labelling extended to the interlobular arteries in sodium depleted kidney. 6. These results indicate that autoradiography combined with the in vitro binding of radiolabelled renin inhibitors may provide a useful tool to measure active and prorenin renin and thereby study the physiological regulation of renin in the kidney.     

Inhibitory effect of a new steroidal saponin,OSW-1, on ovarian functions in rats

1. This study was undertaken to determine the effects of OSW-1 (3β, 16β, 17α-trihydroxycholest-5-en-22-one 16-O-(2-O-4-methoxybenzoyl-β-D-xylopyranosyl)-(1→3)-(2-O-acetyl-α-L-arabinopyranoside)) on the pituitary-ovarian system and the functions of aortic smooth muscle.
2. A single s.c. injection of OSW-1 (9 μg kg⁻¹) on the morning of pro-oestrus inhibited the occurrence of the expected next pro-oestrus, whereas administration of OSW-1 at a dose of 4.5 μg kg⁻¹ did not affect the oestrous cycle. OSW-1 treatment on the day of dioestrus-l did not affect the oestrous cycle.
3. At doses of 4.5 and 9 μg kg⁻¹ OSW-1, the serum oestradiol (E₂) levels at the expected next pro-oestrus were significantly lower than in control (pro-oestrus). The serum luteinizing hormone (LH) levels 4 days after 9 μg kg⁻¹ OSW-1 treatment were also markedly lower than those of control. OSW-1 (4.5 μg kg⁻¹) did not affect the levels of inhibin, progesterone and gonadotrophins on the same day.
4. OSW-1 did not inhibit the preovulatory LH surge which occurs on the afternoon of pro-oestrus day.
5. The expression of mRNA coding for the cholesterol side chain cleavage cytochrome P-450 (p450scc), an ovarian steroidal limiting enzyme, was suppressed at 24 and 96 h after OSW-1 treatment.
6. Administration of OSW-1 (9 μg kg⁻¹) tended to reduce the relaxation of isolated thoracic aorta ring preparations induced by acetylcholine, while there was no difference in the relaxation induced by sodium nitroprusside.
7. Our results show that OSW-1 inhibits ovarian E₂ secretion and that the decrease in E₂ secretion may contribute to its effects on the oestrous cycle and the sensitivity of the thoracic aorta to relaxation. The decrease in the levels of ovarian steroids induced by OSW-1 may be due to its direct inhibitory action on the gene expression of the steroidal enzyme and on the proliferation of granulosa cells in the ovary.

     

Steroidogenesis-disrupting compounds can be effectively studied for major fertility-related endpoints using in vitro cultured mouse follicles

Biologically relevant bioassays are needed to test various endocrine disrupters (EDs). A mouse follicle culture model could allow measuring steroidogenic enzyme function in combination with oocyte growth and meiotic maturation using routine methodology. Three steroidogenesis-disrupting ‘model’ chemicals were tested; vorozole (VOR), aminoglutethimide (AMG), and ketoconazole (KCZ). Along with visual assessment of follicular growth, differentiation and oocyte growth and maturation by conventional light microscopy, steroid secretion measurements allowed to confirm literature findings from in vivo animal studies and more complex in vitro tests. The bioassay was applied for a dose–response study of mono(2-ethylhexyl)phthalate (MEHP), a chemical known to disrupt several steroidogenic enzymes. This bioassay was able to confirm an increased inactivation of E₂ to E₁ and an induced precocious progesterone increase, implying that MEHP can disrupt follicle differentiation and impact the reproductive axis. This in vitro ovarian model allows to reduce animal use by performing synchronous culture of large numbers of early preantral ovarian mouse follicles and is informative on multiple fertility-related endpoints.     

Effect of angiotensin converting enzyme inhibition on the menstrual cycle of hypertensive women.

Angiotensin II was reported to play a key role in ovulation in rats and it seems also to be involved in the regulation of LH release. Thus, we studied the effect of chronic ACE inhibition on the menstrual cycle, measuring daily plasma estradiol, progesterone, LH and FSH, and renin and prorenin before and during the third month of treatment with enalapril (10 mg b.i.d.) in 10 mild essential hypertensive women. Blood pressure was normalized by treatment. The cyclical changes of steroids and gonadotrophins were unaffected in their temporal relationships and in the magnitude of their variation during the experimental cycle compared with the basal cycle. A synchronization of plasma prorenin with the other hormones was seen both before, as previously reported, and during enalapril treatment. Our data show that peripheral blockade of angiotensin I conversion does not affect the pituitary guidance of the ovarian hormonal response or the ovarian prorenin release during the menstrual cycle. Our data are in agreement with the hypothesis that circulating angiotensin II does not play a key role in the human fertility process and that hydrophilic ACE inhibitors can be safely used in the treatment of hypertensive women of reproductive age.     

Impairment of ovarian follicular development caused by titanium dioxide nanoparticles exposure involved in the TGF-β/BMP/Smad pathway

Titanium dioxide nanoparticles (TiO₂ NPs) have been shown to induce reproductive system damages in animals. To better underline how TiO₂ NPs act in reproductive system, female mice were exposed to 2.5, 5, or 10 mg/kg TiO₂ NPs by gavage administration for 60 days, the ovary injuries, follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels as well as ovarian follicular development-related molecule expression were investigated. The results showed that TiO₂ NPs exposure resulted in reduction of ovary weight and inhibition of ovarian follicular development. Furthermore, the suppression of follicular development was demonstrated to be closely related to higher FSH and LH levels, and higher expression of activin, follistatin, BMP2, BMP4, TGF-β1, Smad2, Smad3, and Smad4 as well as decreased inhibin-α expression in mouse ovary in a dose-dependent manner. It implies that the impairment of ovarian follicular development caused by TiO₂ NPs exposure may be mediated by TGF-β signal pathway.     

Studies on reproductive endocrine function in rats treated with monosodium l-glutamate early in life

Repeated s.c. injections of high doses of monosodium-l-glutamate (MSG) to neonatal female rats caused precocious puberty, disturbed oestrous cycle and small ovaries and pituitary. Pituitary LH and FSH were low but the basal serum levels were unchanged from control values. Serum E₂ level was significantly low in the early stages. Cyclic regulation of gonadotrophin release and follicular maturation was inadequate. Pituitary-gonadal function in males was less affected. Females treated with high doses of MSG as infants showed normal onset of puberty and regular oestrous cycles, but subsequent earlier oestrous cycle irregularity was observed than in controls. Gonadal weights in both sexes were slightly reduced. Serum hormone levels and the pituitary contents were not changed from those of controls except for reduced FSH. Males and females, given subneurotoxic doses of MSG, or fed large amounts of MSG ad libitum, presented no abnormalities. MSG administration therefore induces marked abnormalities in reproductive endocrine function after maturation only when injected parenterally, early in postnatal life, in repeated, very large doses.     

β2-ADRENORECEPTOR BINDING SITES IN CIRCULAR AND LONGITUDINAL MYOMETRIAL LAYERS OF THE VIRGIN GUINEA-PIG: THE INFLUENCE OF OVARIAN STEROIDS

• 1 Membranes prepared from both circular and longitudinal muscle layers of the uteri of two groups of virgin adult guinea-pigs were used to study the influence of ovarian steroids upon β-adrenoreceptor binding sites. Animals were (i) treated for 14 days with oestradiol cypionate, beginning on day 9–10 of the oestrous cycle; or (ii) treated as in (i) and in addition, with progesterone for the last four days of oestradiol administration.
• 2 (-)-[¹²⁵I]-iodocyanopindolol ([¹²⁵I]-CYP) was used to determine the numbers and characteristics of β-adrenoreceptor bnding sites in the four membrane preparations. In all cases, binding displayed characteristics of a saturable bimolecular reaction; the estimates of receptor site density (B_Max; 0.26-0.33 pmol g^?1 wet weight) were similar in all four preparations, as were those of binding affinity K_D; 18–31 pmol 1^?1).
• 3 The mean negative logarithms of apparent dissociation constants (pK_D) for the inhibition of specific [¹²⁵I]-CYP binding by ICI118, 551 (β₂-adrenoreceptor selective antagonist) ranged from 8.4 to 8.6; and those for L 643, 717-01J10 (pradrenoreceptor selective antagonist) were 5.7-6.2. Thus the [¹²⁵I]-CYP binding sites in all four membrane preparations displayed the characteristics expected of homogeneous populations of adrenoreceptors of the β₂-subtype. The pK_D values for isoprenaline were also similar in each type of membrane preparation (5.8-6.0).
• 4 It is concluded that the clearcut differences in the contractile responsiveness, to adrenoreceptor agonists, of the circular and longitudinal myometrial preparations from oestrogen-treated guinea-pigs are not due to differences in the numbers, subtype or binding affinities of β-adrenoreceptor binding sites. Moreover, enhancement of the inhibitory potencies of β-adrenoreceptor agonists, in preparations of longitudinal myometrium from oestrogen-primed guinea-pigs treated with progesterone, is not due to an effect of progesterone upon β-adrenoreceptor binding sites.

     

Involvement of adrenoceptors in the ovarian vascular pedicle in the regulation of counter current transfer of steroid hormones to the arterial blood supplying the oviduct and uterus of pigs

1. On Day 10 of the oestrous cycle in pigs, after laparotomy noradrenaline (NA), methoxamine (α₁-adrenomimetic, M), Prazosin (α₁-adrenolytic, Pr) in total doses of 4 μmol, and saline were infused (10 min) into the superficial layer of mesovarium on both sides of the ovarian pedicle vasculature, close to the ovary.
2. Blood flow in the ovarian artery, heart rate and progesterone (P₄) and androstenedione (A₄) secretion from the ovary and their concentrations in the ovarian venous effluent, as well as the concentrations of P₄ and A₄ in the blood supplying the oviduct and the uterus, were determined.
3. A significant increase of P₄ and A₄ secretion after NA and M infusion and increased concentrations of P₄ and A₄ in the ovarian venous effluent were found, but these changes did not influence the counter current transfer of hormones from the venous effluent into arterial blood supplying the oviduct and the uterus.
4. Infusion of Pr caused a significant decrease of P₄ and A₄ secretion and their concentrations in the ovarian venous effluent and significantly increased A₄ concentration in the blood supplying the oviduct and uterus.
5. The results indicate that stimulation of α₁-adrenoceptors in the area of ovarian vasculature did not influence, whereas block of α₁-adrenoceptors affected, the local concentration of steroid hormones in the blood supplying the oviduct and the part of the uterus proximal to the ovary, despite the changes in the concentrations of steroid hormones in the ovarian effluent.

     

还少胶囊联合戊酸雌二醇片/雌二醇环丙孕酮片复合包装治疗卵巢早衰的临床研究

目的探讨还少胶囊联合戊酸雌二醇片/雌二醇环丙孕酮片复合包装治疗卵巢早衰的临床疗效。方法选取2015年2月—2017年6月南开大学附属医院(天津市第四医院)收治的132例卵巢早衰患者,随机分为对照组和治疗组,每组各66例。对照组患者给予戊酸雌二醇片/雌二醇环丙孕酮片复合包装,1片/d,连续服用21 d。于撤退性出血第5天重复使用,连续治疗4个月经周期。治疗组在对照组治疗基础上口服还少胶囊,5粒/次,3次/d。有月经来潮者,于月经第5天服用,连续治疗4个月经周期。观察两组的临床疗效,比较两组治疗前后平均卵巢直径(MOD)、窦卵泡个数(AFC)、卵巢动脉收缩期峰值流速(PSV)、子宫内膜厚度、雌二醇(E_2)、卵泡刺激素(FSH)、黄体生成素(LH)、CD~(3+)、CD~(4+)、CD~(8+)、CD~(4+)/CD~(8+)比值的变化情况。结果治疗后,对照组和治疗组的总有效率分别为83.33%、95.45%,两组比较差异有统计学意义(P0.05)。治疗后,两组MOD、AFC、PSV、子宫内膜厚度、E_2、CD~(3+)、CD~(4+)、CD~(4+)/CD~(8+)比值均较治疗前显著增加,但FSH、LH、CD~(8+)显著降低,同组治疗前后比较差异具有统计学意义(P0.05);治疗后,治疗组MOD、AFC、PSV、子宫内膜厚度、E_2、CD~(3+)、CD~(4+)、CD~(4+)/CD~(8+)比值高于对照组,FSH、LH、CD~(8+)低于对照组,两组比较差异有统计学意义(P0.05)。结论还少胶囊联合戊酸雌二醇片/雌二醇环丙孕酮片复合包装治疗卵巢早衰疗效显著,可有效缓解临床症状,调节性激素水平,促进卵巢功能恢复,增强细胞免疫功能,具有一定的临床推广应用价值。     

β-Adrenergic regulation of renin expression in differentiated U-937 monocytic cells

Previous studies from our laboratories demonstrated that human decidual macrophages and peripheral mononuclear cells express renin. In the present study, we found that U-937 monocytes, induced to differentiate into macrophage-like cells by treatment with phorbol dibutyrate (PDBU), express renin mRNA and release renin (95% of which is in the form of prorenin). Treatment of these PDBU-exposed cells with dibutyryl-cAMP (1 mM) caused a 20-fold increase in renin mRNA and a 10-fold increase in prorenin release. Forskolin (10 μM), an activator of adenylyl cyclase, and terbutaline (100 μM), a β₂-adrenergic agonist known to increase cAMP levels, also increased renin mRNA and prorenin release. The secretory response to terbutaline was potentiated by the type IV cyclic AMP-phosphodiesterase (PDE) inhibitor Ro 20–1724 (50 μM). Angiotensin II agonist inhibited the stimulatory effect of terbutaline on renin secretion as did the cytokines tumor necrosis factor-α and lipopolysaccharide plus interferon-γ. Since other studies have shown that U-937 cells possess β₂-adrenergic receptors and express mainly the type IV PDE, the present findings strongly suggest that β-adrenergic receptors in mononuclear cells are coupled to renin expression via the cAMP transduction pathway. The results support a possible role for the renin-angiotensin system in macrophage function and suggest potential autocrine regulatory mechanisms in prorenin expression.     

Effect of exogenous female sex-steroid hormones on β2-adrenoceptors in healthy males

Objective: We have previously demonstrated that exogenous progesterone, but not oestrogen, up-regulated lymphocyte β₂-adrenoceptors (β₂-AR) when given during the follicular phase in healthy females. In the present study, we were interested to see whether this facilitatory effect of female sex-steroid hormones could be demonstrated in healthy males. Methods: Nine healthy male volunteers with a mean age of 24 years completed this randomised, double-blind, placebo-controlled, cross-over study. They were randomised to receive either oral placebo, oestradiol valerate (4 mg) or medroxyprogesterone (40 mg). The study medication was given in two divided doses 12 h apart. Subjects attended the laboratory at baseline (T₀ is baseline), 1 h after ingestion of the second dose of study medication (T₁) and 24 h later (T₂₄). At each visit, 60 ml of peripheral blood was withdrawn for measurement of serum oestradiol, progesterone and testosterone levels, and for lymphocyte β₂-AR parameters; density (B_max), binding affinity (K_d) and maximal cyclic AMP response to isoprenaline (E_max). Results: Baseline levels of sex-steroid hormones were comparable for each of the treatment periods. Serum oestradiol levels increased significantly, twofold, 1 h after ingestion of oestradiol but there was no significant change in levels of serum progesterone and testosterone. Lymphocyte β₂-AR parameters following treatment with oestradiol and progesterone did not change significantly from baseline and were not different from placebo. Conclusion: In contrast to previously reported effects in women, female sex-steroid hormones did not appear to have any significant facilitatory effects on lymphocyte β₂-AR parameters when given exogenously to healthy males. This lack of effect may be due either to the absence of receptors for female sex hormones in β₂-AR or to reduced efficacy of female hormone-receptor coupling in male lymphocytes. Received: 22 October 1996 / Accepted in revised form: 17 January 1997