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1.
Interleukin-1β (IL-1β) may regulate ovarian physiology. In this study, the influence of IL-1β on secretory activity within the corpora lutea (CL) of cyclic and gravid pigs was determined in vitro during different stages of the CL lifespan, e.g. on Days 10-11, 12-13 and 15-16 of the oestrous cycle and pregnancy. IL-1β (10 ng/ml) increased prostaglandin E2 (PGE2) secretion from CL of the cyclic and gravid pigs during studied days of the oestrous cycle and pregnancy. Increase (P < 0.05) of prostaglandin F2α (PGF2α) in IL-1β-treated CL was demonstrated only on Days 10-11 of the oestrous cycle. More potent stimulatory effect of IL-1β on PGE2 than PGF2α secretion resulted in the enhancement of the PGE2:PGF2α ratio in cyclic and early pregnant CL. IL-1β increased (P < 0.05) progesterone (P4) secretion only in gravid CL and had no effect on oestradiol-17β (E2) release. Expression of cyclooxygenase-2 (COX-2) mRNA was stimulated (P < 0.05) in IL-1β-treated cyclic and gravid CL. Expression of prostaglandin synthase mRNAs in response to IL-1β did not increase. In conclusion, IL-1β modulates PGE2, PGF2α and P4 secretion from porcine CL, depending on luteal stage and the surrounding hormonal milieu. The cytokine may act locally in porcine CL for luteotrophic support throughout the PGE2-mediated synthesis and secretion.  相似文献   

2.
In this study we measured protein concentrations of insulin-like growth factor (IGF)-I and IGF binding proteins (IGFBPs) 2-5 in porcine corpora lutea (CLs) throughout the estrous cycle (Experiment 1), and examined the effects of IGFBP-3 and IGFBP-3 antibody (AB) on luteal progesterone (P4) secretion in vitro (Experiment 2). For Experiment 1, (CLs) and serum were collected on days (D) 4, 7, 10, 13, 15 and 16 of the estrous cycle (n = 5 animals per day). IGF-I was extracted from CLs and sera, and measured by radioimmunoassay (RIA). IGFBPs were measured in CLs by ligand blots. For Experiment 2, CLs (from Experiment 1) were enzyme dissociated and luteal cells cultured (24 h) in Medium 199 (M199) containing (0-500 ng/ml) IGFBP-3 (+/-IGF-I; 100 ng/ml), or (0-10 microg/ml) IGFBP-3 AB. P4 in media was measured by RIA. In Experiment 1, luteal IGF-I concentrations (ng/g tissue) were maximal on day 4 and gradually decreased thereafter. Serum IGF-I concentrations (ng/ml) were highest on days 4 and 7, compared with days 10-15. Peak levels of luteal IGFBP-3 were also seen on days 4 and 7 of the cycle. Luteal IGFBP-2 concentrations showed a tendency to increase on day 16 (P < 0.05 versus day 10), but no significant changes in IGFBP-4 or -5 were seen. In Experiment 2, IGFBP-3 (w IGF) inhibited the steroidogenic actions of IGF-I, but had no significant actions alone (IGFBP-3 w/o IGF). Finally, IGFBP-3 AB stimulated P4 secretion on days 4 and 7, but not on days 10-16. We conclude that IGFBP-3 inhibits IGF-I actions in the porcine CL.  相似文献   

3.
The corpora lutea (CL) are endocrine glands that form in the ovary after ovulation and secrete the steroid hormone, progesterone (P4). P4 plays a critical role in estrous and menstrual cycles, implantation, and pregnancy. The incomplete rodent estrous cycle stably lasts 4–5 days and its morphological features can be distinguished during each estrous cycle stage. In rat ovaries, there are two main types of CL: newly formed ones due to the current ovulation (new CL), and CL remaining from prior estrous cycles (old CL). In the luteal regression process, CL were almost fully regressed after four estrous cycles in Sprague-Dawley rats. P4 secretion from CL in rodents is regulated by the balance between synthesis and catabolism. In general, luteal toxicity should be evaluated by considering antemortem and postmortem data. Daily vaginal smear observations provided useful information on luteal toxicity. In histopathological examinations, not only the ovaries and CL but also other related tissues and organs including the uterus, vagina, mammary gland, and adrenal glands, must be carefully examined for exploring luteal changes. In this review, histological and functional characteristics of CL in rats are summarized, and representative luteal toxicity changes are presented for improved luteal toxicity evaluation in preclinical toxicity research.  相似文献   

4.
Some studies in mammalian species recently demonstrated that tumor necrosis factor (TNF)-alpha plays an important role for corpus luteum (CL) function by way of apoptosis during the estrous cycle. The objectives of this study were to clarify the induction of apoptosis in cultured porcine luteal cells by TNF-alpha treatment. Luteal cells prepared from porcine ovaries collected from crossbred mature gilts on Days 10-14 of the estrous cycle were isolated and examined as follows: 1) Flow cytometric analysis was carried out to determine apoptosis in cultured luteal cells. 2) Single-cell gel electrophoresis (comet assay) was performed to investigate apoptotic DNA fragmentation in luteal cells. The results of the flow cytometric analysis and comet assay demonstrated coincidentally that TNF-alpha induces DNA fragmentation in luteal cells causing apoptosis. These results revealed that TNF-alpha is an inducing factor of apoptosis in luteal cells.  相似文献   

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8.
Corpus luteum (CL) function following spontaneous or prostaglandin F2 alpha-induced estrus was studied in 27 Brahman cows and 16 Brahman heifers. Females received one injection of 0, .38, .75 or 2.25 mg alfaprostol (PGF)/100 kg body weight at 12 +/- .1 d post-estrus. Serum progesterone (P4) during CL formation, following the subsequent estrus, increased over time (P less than .001, 1 to 13 d post-estrus) and was influenced by PGF (P less than .06) and a PGF X time interaction (P less than .02). Low serum P4 concentrations were observed during CL formation (d 3, 4, 10, and 12) in cattle that received 2.25 mg PGF. On d 13 post-estrus, CL were removed. No differences in weight, P4 content or number of luteinizing hormone (LH) receptors was evident between CL derived 13 d following spontaneous vs PGF-induced estrus. The CL formed following PGF-induced estrus had fewer large (P less than .002) and small (P less than .09) luteal cells in heifers, a lower (P less than .008) percentage composition of histological type I and II luteal cells in cows and a lower (P less than .0001) in vitro P4 response to LH in both cows and heifers when compared with the CL formed following spontaneous estrus. Cows had heavier (P less than .002) CL with a higher (P less than .05) number of large cells, a higher (P less than .09) percentage composition of histological cell type I and II luteal cells, and a higher (P less than .03) in vitro basal and stimulated P4 secretion. It is concluded that 1) the events associated with artificially shortening the estrous cycle with PGF altered subsequent CL function in Brahman females; 2) cows had heavier CL, which were composed of more steroidogenically functional luteal cells, than heifers and 3) age of animal interacted with PGF to alter subsequent CL function in Brahman females.  相似文献   

9.
Factors which induce the corpus luteum persistent (CLP) creation in animal ovaries are located in the hypothalamic-pituitary-ovarian axis and also in the uterus. In cows and likewise in others animals, various mediators of inflammatory reaction are released, mainly proinflammatory cytokines from inflamed uterus into the blood and lymph. Afterwards the cytokines cross the blood-brain barrier, and though the brain mediators alter the hormonal profile and amplitude pulses of the hormones release in the hypothalamus and the pituitary. Until it is known, that cytokines: IL-1, IL-2, IL-6, TNF-alpha and also IFN-alpha, administered into the median eminence, cause an increase in corticotrophin-releasing hormone (CRH) and adrenocorticotropic hormone (ACTH) concentrations and decrease in the pituitary gland hormones secretion. The immune system, represented in the corpora lutea (CL) by numerous macrophages/monocytes, limphocytes and neutrophils plays an important role in the luteolysis process. The stimulating factor of the infiltration of these cells is an increased PRL level. The preovulatory increase in PRL level regulates the number of macrophages in newly-formed CL and later influences the number of these cells in the luteolysis period. The pulsatory release and high levels of the hypophyseal oxytocin (OT) and uterine PGF2alpha ensure the beginning and the normal course of the luteolysis period. The cytokines decrease OT concentration and disorder its pulsatory release from the pituitary. In these circumstances the quantity of the uterine PGF2alpha reaching ovaries, is insufficient to begin luteolysis. In the inflamed uterus, the elevation of PGE2 and PGI2 synthesis takes place. Both prostaglandins cause smooth uterine muscles relaxation and the dilatation of blood and lymph vessels in this organ. In these conditions, the blood and lymph outflow from the uterus is several times slower than in the control animals. The secretion of P4 and E2 from CLP, in comparison with control animals, is significantly lower. Decreased P4 concentration during the luteal phase of the estrous cycle, and E2 in the initiation of the luteolysis period, may cause the insufficient preparation of the endometrium for hypophyseal OT activity. Finally, we can assume that the creation of the CLP in the animal ovary is an exceptionally complex and not yet fully understood process.  相似文献   

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11.
Polychlorinated biphenyls (PCBs) due to their lipophilic properties can be easily accumulated in animal and human body and elicit diverse effects causing impairment of reproductive processes. Since these compounds were not be able to affect directly the luteal steroidogenesis, the aim of the present study was to verify hypothesis that PCBs can impair the effect of LH on the secretory function of luteal cells. Bovine luteal cells from different stages of the oestrous cycle (days 1-5, 6-10, 11-15 and 16-18) were exposed for 72h to various congeners of PCBs (PCB 126, PCB 77 and PCB 153) at the doses of 1, 10 or 100 ng/ml, in the presence or absence of LH (100 ng/ml), to determine the possible effect of these compounds on progesterone (P4) and ovarian oxytocin (OT) secretion. Only PCB 77 on days 1-5 and 16-18 increased P4 secretion. All PCBs decreased LH-simulated secretion of P4 from luteal cells obtained from all days of luteal phase. Dioxin-like congener (PCB 126) inhibited (P<0.05) the most evidently LH effect on P4 secretion. All congeners, except the lower doses of PCB 126, increased (P<0.05) OT secretion. They can also increase LH-stimulated secretion of OT, but the effect was dependent on the congener used and on the phase of oestrous cycle. On days 1-5 and 10-15, PCB 126 diminished LH-stimulated effect on OT secretion from luteal cells. PCB 77 (mimickig both dioxin and estradiol effect) in the higher doses, amplified effect of LH-stimulated OT secretion, while on all other days it diminished LH influence. PCB 153, which has estrogen-like properties, amplified LH effect on OT secretion during all studied days of the cycle. We conclude that PCBs (supposedly via estrogen and arylhydrocarbon - AhR receptor) may directly affect LH-stimulated function of CL. This does not appear to be a direct adverse effect on luteal steroidogenesis, but rather indirect on OT secretion from or within CL.  相似文献   

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13.
To determine the physiological significance of tumour necrosis factor‐α (TNFα) in the regulation of luteal functions in pig, this study was conducted to identify the presence of functional TNFα receptors in porcine corpora lutea (CL) throughout the oestrous cycle and the early gestation. The CL were isolated from pigs on days 4, 6, 8, 12 or 15 of the oestrous cycle (n=3; day 0 = oestrus) and days 15, 20 or 25 of gestation (n=3; day 0 = mating). A Scatchard analysis revealed the presence of a high‐affinity binding site for TNFα in all samples (dissociation constant; 2.7 ± 0.51 to 5.8 ± 0.50 nM ). The concentration of TNFα receptors was higher on day 15 of the oestrous cycle than on days 4 and 8 of the oestrous cycle (p < 0.05). Furthermore, TNFα receptor concentrations in the CL on days 15, 20 and 25 of gestation were significantly lower than on day 15 of the oestrous cycle (p < 0.05). On day 9 of the oestrous cycle, exposure of cultured luteal cells to 0.06–60 nM TNFα stimulated prostaglandin (PG) F2α and PGE2 secretion in a dose‐dependent manner (p < 0.05). These results indicate that functional TNFα receptors are present in the porcine CL throughout the oestrous cycle and early gestation, and suggest that TNFα plays one or more physiological roles in regulating CL function throughout the oestrous cycle and the early gestation period. In addition, TNFα receptor concentration in the CL of the late luteal stage (day 15) of the oestrous cycle was higher than on the respective day in the early pregnant pig, suggesting that TNFα plays a role in accomplishing luteolysis in the porcine CL.  相似文献   

14.
The objective of the present study was to investigate the influence of prostaglandin F(2alpha) (PGF (2alpha)) and nitric oxide (NO) on production of steroids and PGs by culturing bovine luteal cells obtained from ovaries on days 8-12 of the estrous cycle with a nitric oxide (NO) donor (Spermine NONOate), and a NO synthase inhibitor (N(G)-nitro-L-arginine methyl ester dihydrochloride: L-NAME). When the cells were exposed for 24 h to PGF(2alpha) (10(-7)-10(-5) M), production of progesterone (P(4)) increased significantly at all doses used (P<0.05). Moreover, PGF(2alpha) stimulated PGF(2alpha) production (P<0.01), depressed testosterone (T) production (P<0.05), but did not affect synthesis of prostaglandin E(2) (PGE(2)). Spermine NONOate decreased P(4) production to 66%, 47% and 34% of the control concentration after treatment with 10(-5) M, 10(-4) M and 10(-3) M, respectively, but did not affect T production, and increased PGF(2alpha) synthesis (P<0.05) and PGE(2) (P<0.01) at all doses used. L-NAME increased production of P(4) (P<0.01) but did not affect (P>0.05) secretion of T, PGF(2alpha) and PGE(2). Estradiol-17beta (E(2)) was detectable on the level of sensitivity of assay and was not significantly altered by any treatments. The overall results suggest that PGF(2alpha) and NO produced locally in bovine CL play roles in the regulation of the secretory function of the bovine CL as auto/paracrine factors.  相似文献   

15.
Prostaglandin F2alpha (PGF2alpha) is the primary luteolysin in the cow. During the early luteal phase, the corpus luteum (CL) is resistant to the luteolytic effect of PGF2alpha. Once mature, the CL becomes responsive to PGF2alpha and undergoes luteal regression. These actions of PGF2alpha coincide with changes in luteal blood flow (BF): PGF2alpha has no effect on BF in the early CL, but acutely increases BF in the peripheral vasculature of the mature CL within 30 min of PGF2alpha injection. During spontaneous luteolysis, luteal BF increases on Days 17-18 of the estrous cycle, prior to any decrease in plasma progesterone (P). The increase in luteal BF is synchronous with an increase in plasma PGFM levels, suggesting that pulsatile release of PGF2alpha from uterus stimulates the increase in luteal BF. Serial biopsies of these CL showed that mRNA expression for endothelial nitric oxide synthase (eNOS) together with endothelin-1 (ET-1) and angiotensin converting enzyme (ACE) increases on Days 17-18 when the luteal BF is elevated. On Day 19 when plasma P level firstly decreases, eNOS mRNA returns to the basal level whereas ET-1 and ACE mRNA remains elevated. Cyclooxygenase-2 (COX-2) mRNA expression increases on Day 19. In support of these data, an in vivo microdialysis study revealed that luteal ET-1 and angiotensin II (Ang II) secretion increases and precedes PGF2alpha secretion during spontaneous luteolysis. In conclusion, we show for the first time that an acute increase of BF occurs in the peripheral vasculature of the mature CL together with increases in eNOS expression and ET-1 and Ang II secretion in the CL during the early stages of luteolysis in the cow. We propose that the increase in luteal BF may be induced by NO from large arterioles surrounding the CL, and simultaneously uterine or exogenous PGF2alpha directly increases ET-1 and Ang II secretion from endothelial cells of microcapillary vessels within the CL, thereby suppressing P secretion by luteal cells. Taken together, our results indicate that an acute increase in luteal BF occurs as a first step of luteolysis in response to PGF2alpha. Therefore, local BF plays a key role to initiate luteal regression in the cow.  相似文献   

16.
The aim of the present study was to investigate the influence of progesterone (P4), its precursor (pregnenolone; P5) and metabolite (17beta-hydroksyprogesterone; 17betaOHP4) on secretory function of bovine luteal cells on days 6-10 of the estrous cycle and on intracellular Ca2+ mobilization. The luteal cells were pre-incubated for 24 h and after change of medium they were incubated for 30 min with P5 and 17betaOHP4 (10(-5) each). Next, the medium was supplemented with LH (100 ng/ml), noradrenaline (NA; 10(-5) M) and prostaglandin (PG)E2 (10(-6) M), the cells were incubated for further 4 h and the medium was collected for P4 determination. Another set of luteal cells (5x10(4)/well) was incubated with P4, P5 and 17betaOHP4 at the dose of 10(-5) M each for 30 min and intracellular Ca2+ mobilization was measured every 5 s three times before and for 60 s after cells stimulation with LH, NA and PGE2. Metabolite of P4 did not affect the stimulatory effect of LH, PGE2 and NA on P4 secretion to the medium. Whereas all used steroids reduced calcium release from small but not from large luteal cells. It is suggested that steroids could temporary impair effect of luteotropins on the luteal cells via non-genomic way.  相似文献   

17.
Effect of polychlorinated biphenyles (PCBs) on viability and secretory function of luteal and granulosa cells from mature cows was studied. Luteal cells from corpora lutea of different developmental stages and granulosa cells from follicles of >1 cm< in diameter were used. Neither individual congeners (PCB-126, -77, -153) nor mixture of PCBs Aroclor Ar) 1248 at the dose of 1, 10 or 100 ng/ml affected the viability of cells (P>0.05) compared to control after 72 h of incubation. PCBs markedly increased (P<0.05-0.001) oxytocin (OT) secretion from granulosa cells. This effect was the most evident when granulosa cells from follicles <1 cm diameter was treated with PCB-77 which is assumed to stimulate both arylhydrocarbon receptor (AhR) and estradiol (E2) receptor. Even the lowest dose of this compound (1 ng/ml) outranged the effect produced by cortisol (10(-5)M) used as positive control. There was marked effect (P<0.05-0.001) of PCBs on luteal cells from days 6-15 of the estrous cycle. However, influence of PCBs on OT secretion from luteal cells on day 1-5 and 16-18 of the estrous cycle was less evident. Again, PCB-77 was the most efficient stimulator of OT secretion. While the lowest effect was found after treatment of cells with PCB-126 which has dioxin-like properties. It can be assumed that diverse effect of PCBs on female reproduction largely results from the influence of these compounds on ovarian OT secretion. Since both synthesis and secretion of ovarian OT in bovine do not markedly depend on estradiol, some alternative cellular pathways of PCBs on ovary function are suggested.  相似文献   

18.
Cell-to-cell interaction via cell contact-dependent pathway is essentially important for maintenance and regulation of corpus luteum (CL) integrity and its physiological actions. The objective of the present study was to evaluate the mRNA expression of the cell adhesion molecules (CAMs) that are constituent factors of gap junctions [connexin (Cx) 43] and adherence junctions (VE-, E-, N-cadherin) in two types of endothelial cells from the mid CL and in CL tissue during the estrous cycle and PGF(2alpha)-induced luteolysis in the cow. Specific mRNA expression for Cx43 and N-cadherin was detected in cytokeratin-positive (CK+) and cytokeratin-negative (CK-) luteal endothelial cells (EC) and fully luteinized granulosa cells (LGC). E-cadherin mRNA was expressed in CK+EC and LGC, but not in CK-EC. VE-cadherin mRNA was expressed in both CK+ and CK-EC. During the estrous cycle, Cx43 mRNA expression was significantly lower in the regressing CL. VE-cadherin expression also tended to increase in the mid CL and increased significantly in the regressing CL. E-cadherin mRNA expression was higher in the early and late CL than in the mid- and regressing CL. N-cadherin mRNA expression gradually increased from the early to late CL followed by a decrease in the regressing CL. During PGF(2alpha)-induced luteolysis, Cx43 mRNA expression appeared to increase, and VE-cadherin and E-cadherin mRNA significantly increased at 24 h. N-cadherin mRNA expression decreased 2 and 4 h after PGF(2alpha) administration. Collectively, expression of the mRNAs for CAMs was different in the two types of luteal endothelial cells and fully luteinized granulosa cells and changed independently in the CL during the estrous cycle and PGF(2alpha)-induced luteolysis in the cow. The results suggest that CAMs play physiological roles in cell-to-cell communication to regulate both gap and adherence junctions during CL development and regression in the cow.  相似文献   

19.
Ewes were treated with an agonistic analog of luteinizing hormone-releasing hormone (LH-RH) during the luteal phase (d 10) of the estrous cycle. Function of natural and hormonally-induced corpora lutea (CL) was evaluated by measurements of progesterone in sera or luteal tissue. Synthesis and secretion of progesterone by natural CL were not chronically altered by LH-RH. Likewise, there was no in vitro effect of LH-RH on luteal function. When natural CL were surgically removed, newly formed CL functioned at a defective level. Hysterectomy shortly after ovulation did not significantly influence such luteal activity. Induction of ovulation by LH-RH during the follicular phase (d 16) in uterus-intact ewes was followed by normal profiles of luteal secretion of progesterone; serum concentrations of progesterone in animals that were hysterectomized increased in association with development of the CL, but then plateaued at a subnormal level. There were no differences in patterns of secretion of luteinizing hormone in response to LH-RH due to stage of the estrous cycle. Follicles stimulated to ovulate during the luteal phase contained low numbers of steroidogenically-deficient granulosal-lutein cells. These results indicate that: ovine CL are not sensitive to exogenous LH-RH; luteal dysfunction is a consequence of ovulation during the luteal phase, and the etiology of this abnormality appears to be linked with the developmental status of the ovulatory follicle; and CL that are formed from ovulation of a matured follicle begin to develop normally, but then function at a defective rate in the absence of the uterus.  相似文献   

20.
To determine the effect of the hyperadrenal state on corpus luteum (CL) function, we treated intact and adrenalectomized (ADRX) heifers with adrenocorticotropin (ACTH) and hydrocortisone. ACTH treatment of intact heifers by infusion of 1 mg alpha 1-24 ACTH/24 hr from day 2 to 25 of an estrous cycle increased progesterone concentrations in plasma to 4 +/- .52 ng/ml plasma on days 3 to 4. Thereafter, progesterone concentrations declined, indicating severe suppression of CL function as compared to that of control heifers. Similar infusions of ACTH to ADRX heifers did not alter CL function as reflected by progesterone concentrations in jugular plasma. Infusion of ADRX animals with hydrocortisone succinate (100 mg/24 hr) decreased progesterone in plasma during the cycle, with maximum values averaging less than 4 ng/ml, compared to concentrations of 6 to 7 ng/ml in control ADRX heifers. Chronic administration of ACTH or hydrocortisone to heifers caused a decreased progesterone secretion during the luteal phase of the estrous cycle. On the basis of these results, we suggest that stress, as indicated by the hyperadrenal state, could inhibit progesterone secretion by the corpus luteum.  相似文献   

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