Insulin-like Growth Factor I Release Research Articles

MicroRNA miR-125b can suppress ovarian granulosa cell functions: Interrelationships with FSH.

This study aimed to evaluate the involvement of miR-125b and its interrelationship with follicle-stimulating hormone (FSH)in the control of basic ovarian granulosa cell functions. The effect of miR-125b mimics on basic functions of porcine ovarian granulosa cells cultured with and without FSH, and the effect of FSH on the expression of endogenous miR-125b was examined. Expression levels of miR-125b, viability, proliferation (accumulation of PCNA and cyclin B1), apoptosis (accumulation of bax and caspase 3), the accumulation of FSH receptors (FSHR), steroid hormones, insulin-like growth factor I (IGF-I), oxytocin, and prostaglandin E2 release were analysed by reverse transcription-quantitative polymerase chain reaction, Trypan blue exclusion test, quantitative immunocytochemistry, and ELISA. Transfection of cells with miR-125b mimics inhibited cell viability, proliferation, apoptosis, the occurrence of FSHR, progesterone, testosterone, estradiol, and oxytocin release but stimulated prostaglandin E2 output. FSH promoted cell viability, proliferation, steroid hormones, IGF-I, oxytocin, and prostaglandin E2 output and reduced the expression of miR-125b and apoptosis. Furthermore, miR-125b mimics supported the effect of FSH on the release of estradiol, IGF-I, and prostaglandin E2, and inverted FSH influence on cell viability, proliferation, apoptosis, progesterone, and testosterone output. FSH supported both inhibitory and stimulatory action of miR-125b on ovarian cell functions. Present observations indicate that: miR-125b can be involved in the control of basic ovarian functions and that miR-125b and FSH are antagonists in their actions on ovarian cell functions. The ability of FSH to reduce miR-125b expression and the ability of miR-125b mimics to decrease the occurrence of FSHR and to modify FSH effects indicate the existence of the self-inhibiting FSH-miR-125b axis and that miR-125b can mediate the actions of FSH on ovarian cells.

Merging bioresponsive release of insulin-like growth factor I with 3D printable thermogelling hydrogels.

3D printing of biomaterials enables spatial control of drug incorporation during automated manufacturing. This study links bioresponsive release of the anabolic biologic, insulin-like growth factor-I (IGF-I) in response to matrix metalloproteinases (MMP) to 3D printing using the block copolymer of poly(2-methyl-2-oxazoline) and thermoresponsive poly(2-n-propyl-2-oxazine) (POx-b-POzi). For that, a chemo-enzymatic synthesis was deployed, ligating IGF-I enzymatically to a protease sensitive linker (PSL), which was conjugated to a POx-b-POzi copolymer. The product was blended with the plain thermogelling POx-b-POzi hydrogel. MMP exposure of the resulting hydrogel triggered bioactive IGF-I release. The bioresponsive IGF-I containing POx-b-POzi hydrogel system was further detailed for shape control and localized incorporation of IGF-I via extrusion 3D printing for future applications in biomedicine and biofabrication.

The Effects of Alpha-Linolenic Acid on the Secretory Activity of Astrocytes and β Amyloid-Associated Neurodegeneration in Differentiated SH-SY5Y Cells: Alpha-Linolenic Acid Protects the SH-SY5Y cells against β Amyloid Toxicity.

Alzheimer's disease (AD) is the most common neurodegenerative disorder. Amyloid β- (Aβ-) induced mitochondrial dysfunction may be a primary process triggering all the cascades of events that lead to AD. Therefore, identification of natural factors and endogenous mechanisms that protect neurons against Aβ toxicity is needed. In the current study, we investigated whether alpha-linolenic acid (ALA), as a natural product, would increase insulin and IGF-I (insulin-like growth factor I) release from astrocytes. Moreover, we explored the protective effect of astrocytes-derived insulin/IGF-I on Aβ-induced neurotoxicity, with special attention paid to their impact on mitochondrial function of differentiated SH-SY5Y cells. The results showed that ALA induced insulin and IGF-I secretion from astrocytes. Our findings demonstrated that astrocyte-derived insulin/insulin-like growth factor I protects differentiated SH-SY5Y cells against Aβ1-42-induced cell death. Moreover, pretreatment with conditioned medium (CM) and ALA-preactivated CM (ALA-CM) protected the SH-SY5Y cells against Aβ1-42-induced mitochondrial dysfunction by reducing the depolarization of the mitochondrial membrane, increasing mitochondrial biogenesis, restoring the balance between fusion and fission processes, and regulation of mitophagy and autophagy processes. Our study suggested that astrocyte-derived insulin/insulin-like growth factor I suppresses Aβ1-42-induced cytotoxicity in the SH-SY5Y cells by protecting against mitochondrial dysfunction. Moreover, the neuroprotective effects of CM were intensified by preactivation with ALA.

Are the Effects of Oral and Vaginal Contraceptives on Bone Formation in Young Women Mediated via the Growth Hormone-IGF-I Axis?

Purpose: Combined hormonal contraceptive therapy has been associated with negative bone mineral density outcomes that may be route-dependent [i.e., combined oral contraception (COC) vs. contraceptive vaginal ring (CVR)] and involve the hepatic growth hormone (GH)/insulin-like growth factor-I (IGF-I) axis. The objective of the pilot study was to assess the impact of route of contraceptive administration on IGF-I and procollagen type I N-terminal propeptide (PINP) responses to an IGF-I Generation Test. We hypothesized that the peak rise in IGF-I and PINP concentration and area under the curve (AUC) would be attenuated following COC, but not CVR, use.Methods: Healthy, premenopausal women not taking hormonal contraception were recruited. Women were enrolled in the control group (n = 8) or randomly assigned to COC (n = 8) or CVR (n = 8) for two contraceptive cycles. IGF-I Generation Tests were used as a probe to stimulate IGF-I release and were completed during the pre-intervention and intervention phases. Serum IGF-I and PINP were measured during both IGF-I Generation Tests. The study was registered at ClinicalTrials.gov (NCT02367833).Results: Compared to the pre-intervention phase, peak IGF-I concentration in response to the IGF-I Generation Test in the intervention phase was suppressed in the COC group (p < 0.001), but not the CVR or Control groups (p > 0.090). Additionally, compared to the pre-intervention phase, PINP AUC during the intervention phase was suppressed in both COC and CVR groups (p < 0.001), while no difference was observed in the control group (p = 0.980).Conclusion: These data suggest that changes in recombinant human GH-stimulated hepatic IGF-I synthesis in response to combined hormonal contraception (CHC) use are dependent on route of CHC administration, while the influence on PINP is route-independent. Future research is needed to expand these results with larger randomized control trials in all age ranges of women who utilize hormonal contraception.Clinical Trial Registration: www.ClinicalTrials.gov registration NCT02367833.

Relationship between body conditions and environmental contaminants in bovine ovarian cells

The effect of body condition and environmental contaminants on reproductive processes is known; however, it is not known whether basic ovarian cell functions and their response to these contaminants depend on body condition. This study aimed to understand the interrelationships between body conditions and environmental contaminants on ovarian cells. For this purpose, we compared ovarian granulosa cells isolated from cows with an emaciation tendency (body condition score, BCS2 on a scale from 1 to 5) and cows with average body condition (BCS3); proliferation, apoptosis, secretory activity and the response to environmental contaminants were all assessed in the cells. In the 1st series of experiments, ovarian granulosa cells isolated from BCS2 and BCS3 cows were cultured with and without benzene, xylene, and toluene (0.1%). The accumulation of nuclear and cytoplasmic markers of apoptosis (p53 and bax, respectively), a proliferation marker (PCNA), progesterone (P4), and insulin-like growth factor I (IGF-I) was evaluated by Western blot and radioimmunoassay (RIA) experiments. In the 2nd series of experiments, the groups of granulosa cells were cultured with and without mycotoxine deoxynivalenol (DON, 0, 10, 100, or 1000 ng/ml). The secretion of P4 and testosterone (T) was measured by RIA. In comparison to cells from BCS2 animals, ovarian cells isolated from BCS3 cows accumulated higher levels of bax and PCNA but not p53, and they secreted higher amounts of IGF-I but not P4 or T. In cells from BCS2 animals, benzene and xylene promoted p53 accumulation, and toluene reduced the accumulation. In the BCS2 group, all treatments promoted bax and PCNA expression. However, in cells from BCS3 animals, all environmental pollutants inhibited p53, toluene inhibited PCNA but not bax, and xylene did not affect the expression of proliferation or apoptosis markers. In the BCS2 group, P4 was inhibited by xylene, and IGF-I was stimulated by xylene but not by benzene or toluene. Low-dose exposure to DON (10 ng/ml) promoted P4 release from cells from both BCS2 and BCS3 animals, but high-dose exposure to DON (1000 ng/ml) reduced P4 secretion from the cells from BCS2 animals but not from the cells from BCS3 animals. The release of T was inhibited by high-dose exposure to DON (1000 ng/ml), irrespective of the BCS. An emaciation tendency reduces proliferation, apoptosis, and IGF-I release, and it induces or reverses the action of environmental contaminants on ovarian functions. Taken together, these observations demonstrate the effect of body condition and the direct influence of environmental contaminants on basic bovine ovarian functions. Furthermore, they demonstrate for the first time that the response of ovarian cells to environmental contaminants can be regulated by cow body condition.

Role of growth hormone in development and maintenance of follicles and corpora lutea

Growth hormone (GH) is a pituitary hormone that affects animal growth, metabolism, lactation, and reproduction. Many of the effects of GH are mediated by insulin-like growth factor I (IGF-I) which is synthesized in liver and ovary in response to GH. Insulin-like growth factor I synergizes with gonadotrophins (LH and FSH) to stimulate growth and differentiation of ovarian cells. There are species differences in the effects of GH in reproductive biology. In most species, ovarian follicles and corpora lutea are potential sites for GH action because the GH receptor is found within granulosal cells as well as corpora lutea. However, growth hormone does not control ovarian IGF-I in all species and, in ruminants, endocrine IGF-I from liver may be the principal mediator of GH action. In cattle, administration of GH increases the number of small antral ovarian follicles but does not increase the number of large antral (dominant) follicles. Growth hormone may antagonize some aspects of dominant follicular function because dominant follicles are shorter-lived in GH-treated cattle. The corpora lutea has increased growth and steroidogenesis in response to GH. Growth hormone-induced steroidogenesis in cultured granulosal and luteal cells depends on IGF-I release after GH treatment. Bovine and ovine granulosal cells do not release IGF-I in response to GH in vitro and, therefore, are less responsive to GH. These results demonstrate that GH is required for normal reproductive function in ruminant as well as nonruminant species.

The involvement of the phosphorylatable and nonphosphorylatable transcription factor CREB-1 in the control of human ovarian cell functions

The objective of our study was to elucidate the role of the transcription factor CREB-1 in controlling ovarian cell proliferation, apoptosis, and hormone release and the significance of CREB-1 phosphorylation in these processes. Human ovarian granulosa cells were transfected with a gene construct encoding wild-type CREB-1 (CREB-1 WT) or CREB-1 nonphosphorylatable mutant (CREB-1 M1). The expression of total and phosphorylated CREB-1, markers of proliferation (PCNA) and apoptosis (bax), as well as the release of progesterone, oxytocin, prostaglandin F2 alpha (PGF2), prostaglandin E2 (PGE2), and insulin-like growth factor I (IGF-I) were compared by immunocytochemistry, enzyme immunoassay (EIA), and immunoradiometric assay (IRMA). Transfection with CREB-1 WT or CREB-1 M1 increased total CREB-1 expression and proliferation and decreased the release of oxytocin, PGE2, and IGF-I by ovarian cells. CREB-1 M1, not CREB-1 WT, promoted apoptosis and inhibited progesterone output. PGF2 release was inhibited by CREB-1 WT but stimulated by CREB-1 M1 construct. Phosphorylated CREB-1 was undetected in any cell group. These observations confirm the involvement of CREB-1 in the control of ovarian cell proliferation, apoptosis, and steroid hormone release. This is the first demonstration of the involvement of CREB-1 in the regulation of the ovarian non-steroidal hormones such as oxytocin, PGF2, PGE2, and IGF-I. The absence of CREB-1 phosphorylation, similar effects exerted by CREB-1 WT and CREB-1 M1 on cell proliferation and release of oxytocin, PGE2, and IGF-I, and the influence of CREB-1 M1 on apoptosis and progesterone suggest that phosphorylation plays no role in the action of CREB-1 on the majority of analyzed functions of human ovarian cells.

Bioresponsive release of insulin-like growth factor-I from its PEGylated conjugate

PEGylation of protein ligands, the attachment of polyethylene glycol (PEG) polymers to a therapeutic protein, increases therapeutics' half-life but frequently comes at the cost of reduced bioactivity. We are now presenting a bioinspired strategy leading out of this dilemma. To this end, we selected a position within insulin-like growth factor I (IGF-I) for decoration with a PEG30kDa-modified protease-sensitive peptide linker (PSL) using a combination of enzymatic and chemical bioorthogonal coupling strategies. The PSL sequence responded to matrix metalloproteinases (MMP) to provide a targeted release in diseased tissue. The IGF-PSL-PEG conjugate had different binding protein affinity, cell proliferation, and endocytosis patterns as compared to the wild type. Exposure of the conjugate to elevated levels of activated MMPs, as present in inflamed tissues, fully reestablished the wild type properties through effective PSL cleavage. In conclusion, this bioinspired approach provided a blueprint for PEGylated therapeutics combining the pharmacokinetic advantages of PEGylation, while locally restoring the full suite of biological potential of therapeutics.

Cow body condition affects the hormonal release of ovarian cells and their responses to gonadotropic and metabolic hormones

The body condition score (BCS) of cows affects their reproductive efficiency, but the underlying mechanism is unclear. We examined the effect of BCS on the basic ovarian cell functions and their responses to gonadotropic and metabolic hormones. We isolated ovarian cells from cows with a tendency toward emaciation (BCS2) and those with an average body condition (BCS3), and we compared their hormonal release and responses to FSH, leptin, ghrelin, and neuropeptide Y (NPY) added at doses of 0, 1, 10, or 100 ng/mL. Progesterone, testosterone, estradiol, and insulin-like growth factor I (IGF-I) release were evaluated by RIA. No differences were found in progesterone or testosterone release between BCS2 and BCS3 cells; however, ovarian cells from BCS2 cows released more estradiol and IGF-I than cells from BCS3 cows. FSH, ghrelin, and NPY promoted progesterone release in BCS2 cells but had no stimulatory or inhibitory effect on BCS3 cells. In contrast, leptin promoted progesterone release in BCS3 cells and inhibited progesterone release in BCS2 cells. FSH also promoted testosterone release in both BCS2 and BCS3 cells but inhibited progesterone at a low dose in BCS3 cells. Leptin inhibited testosterone release in BCS3 cells but not in BCS2 cells. Estradiol release was promoted by leptin and ghrelin in BCS3 cells; however, it was unaffected by leptin and inhibited by ghrelin in BCS2 cells. IGF-I production was promoted by FSH and inhibited by leptin in both groups. Ghrelin suppressed IGF-I release in BCS2 cells and increased IGF-I release in BCS3 cells. NPY promoted IGF-I release in BCS2 cells but not in BCS3 cells. Our results demonstrate the effects of BCS on ovarian cell estradiol and IGF-I (but not progesterone or testosterone) release, as well as on the responses of ovarian cells to FSH, leptin, ghrelin, and NPY.

Body fat affects mouse reproduction, ovarian hormone release, and response to follicular stimulating hormone

We investigated the effects of body fat content on mouse fecundity, ovarian hormone release, and their response to follicle stimulation hormone (FSH). 4 types of females were produced: lean (group 1), normal (group 2), slightly fat (group 3), and significantly fat (group 4). The body weights, fat content, fertility rate, embryo number produced, retarded and degenerated embryo percentage, the release of progesterone (P4), testosterone (T), and insulin-like growth factor I (IGF-I) by isolated ovaries cultured with and without FSH (1.0IU/mL medium) were evaluated. A gradual increase in body weight and fat contents from groups 1 to 4 was observed. Group 2 had higher fertility rate than those from the other groups. Groups 2 and 3 had fewer retarded and degenerated embryos that those from groups 1 and 4. Embryo production rate was not different among the groups. P4 and T secretion was higher from group 4 than in those from groups 1–3; secretion of IGF-I of group 3 was less than that of groups 1, 2, and 4. FSH promoted ovarian T output in all groups and stimulated ovarian P4 release in groups 1, 3, and 4, but not in group 2. FSH did not affect IGF-I release in any group. Therefore, both malnutrition and overfeeding can affect body weight and fat content in female mice, reducing embryo quality or developmental capacity, but not fertility and embryo production. Excess weight or fat can have stimulatory effects on ovarian P4 and T, but inhibitory effects on ovarian IGF-I release. Both leanness and excess weight or fat can induce the stimulatory action of FSH on ovarian P4.

Metabolic state can define the ovarian response to environmental contaminants and medicinal plants.

Environmental contaminants and medicinal plants can affect reproductive processes. The aim of this study was to investigate the effect of maternal metabolic status on the response of mouse ovaries to the environmental contaminants benzene and xylene, as well as to extracts of the medicinal plant yucca. Ovaries isolated from normal-lean and slightly obese mice were cultured with or without 0.1% benzene or xylene for 24 h. Similarly, ovaries isolated from normal-lean, slightly obese, and significantly obese mice were cultured for 24 h with or without an extract of Yucca shidigera (YS, 10 μ g/mL). We found that the metabolic status did not influence the release of basal progesterone (P4), testosterone (T), or insulin-like growth factor I (IGF-I), but obesity influenced the effects of the environmental contaminants and YS. Benzene reduced P4 output in ovaries from obese but not normal-lean mice; it also reduced IGF-I (but not T) release from ovaries irrespective of the metabolic status. Xylene dramatically increased P4 and T (but not IGF-I) release by ovaries from normal-lean mice, but there were no changes in P4 and only small increases in T output in obese mice. YS increased P4 (but not T or IGF-I) release in normal-lean and slightly obese animal ovaries, whilst significant obesity was associated with a lack of P4 response to YS. Obesity might affect the basal ovarian release of T or IGF-I and increases the sensitivity of ovaries to the action of benzene but decreases their responsiveness to xylene and YS.

Acute Effects of Blood Flow Restriction Resistance Exercise in a Head-down Tilt Position on Growth Hormone and Insulin-like Growth Factor-I Release

Muscular atrophy is common among astronauts while in spaceflight. Growth Hormone (GH) and IGF-I are key hormones with regards to the preservation and growth of skeletal muscle tissue. Low intensity blood flow restriction (BFR) resistance exercise is a newer training modality employed to various populations to enhance muscle growth, yet has not been thoroughly explored in the astronaut population. PURPOSE: To determine the effects of low intensity BFR in a head-down tilt position on the acute release of GH and IGF-I. METHODS: Twelve college-aged males with resistance exercise experience were included in the study. The first of three sessions consisted of informed consent, demographic measurements, and 3RM testing. The two experimental sessions consisted of subjects resting in a -6 deg head-down tilt position for 2 hr. Upon rest completion, while tilted, subjects performed the exercise protocol at 30% of estimated 1RM for the following exercises: close-grip bench press, dumbbell bicep curls, and dumbbell triceps extensions. Exercises were performed with or without BFR (160 mmHg) to the upper arms in a counterbalanced order. Venus blood draws (15 mL) occurred 10 min prior to tilt, 10-15 min post exercise protocol, and 24 hr post exercise protocol and were assayed in duplicate. Two 2 x 2 Factorial ANOVAs with repeated measures were utilized for statistical analysis of the dependent variables (GH, IGF-I). Alpha levels were set at p < 0.05. RESULTS: A significant interaction was found between exercise condition and time for GH (p < .05). No significant interaction was found between exercise condition and time for IGF-I (p > .05). No significant mean difference was observed between BFR and no BFR for IGF-I (p >.05). A significant mean difference was observed pre to post exercise protocol for IGF-I (p < .05). CONCLUSION: A low intensity BFR exercise protocol in a head-down tilt position elicited a large increase in systemic GH concentrations from pre to post exercise. Post exercise GH levels were elevated to a much greater extent with BFR compared to no BFR. A low intensity BFR modality may be beneficial to employ to astronauts while in spaceflight or post spaceflight to help promote muscular tissue growth.

Interrelationships between ovarian follicles grown in culture and possible mediators

Dominance or cooperation between ovarian follicles can determine the number of ovulations and fecundity, but interrelationships between follicles in mono- and poly-ovulatory species and their mechanisms are poorly understood. The goals of this work were to determine the existence and compare the character of mutual influence of cultured ovarian follicles from a mono-ovulatory species (cow) with established follicular dominance with those from a poly-ovulatory species (pig), in which interrelationship between follicles remain unknown, and to examine the role of ovarian cell proliferation, the insulin-like growth factor I (IGF-I)- oxytocin (OT) system, and steroid hormones in mediating interrelationships among ovarian follicles. Bovine and porcine ovarian follicles were isolated and cultured alone and in pairs, and the percentage of growing follicles was calculated. Porcine follicles were cultured alone and in pairs after addition of exogenous OT and IGF-I (100ngmL−1) or inactivation of endogenous OT and IGF-I by antisera against these hormones (1%). Proliferation of porcine follicular cells was assessed by SDS PAGE-Western immunoblotting, the release of IGF-I, progesterone, androstenedione and estradiol by cultured porcine ovarian follicles was analyzed by RIA/EIA. Overall, our observations suggest (1) competition/dominance (mutual suppression of growth) in bovine ovarian follicles, (2) cooperation (mutual support of growth) in porcine ovarian follicles, (3) that this mutual growth of porcine ovarian follicles was caused by the promotion of cell proliferation, (4) that this mechanism was probably not involved in bovine follicular dominance, (5) that communication between both porcine and bovine follicles affects their secretory activity, and (6) that both follicular dominance in cows and cooperation of follicles in pigs can be mediated by either down- or up-regulation of the IGF-I-OT system, which in turn affects follicular steroidogenesis and promotes follicular cell proliferation and follicular growth.

Dietary supplementation of yucca (Yucca schidigera) affects ovine ovarian functions

Yucca (Yucca schidigera) is a popular medicinal plant due to its many positive effects on animal and human physiology, including their reproductive systems. To examine the effect of supplemental yucca feeding on sheep reproduction, including ovarian functions and their hormonal regulators, ewes were fed (or not fed, control) yucca powder (1.5 g/head/day, 30 days). Macromorphometric indexes of the oviduct, ovary, and ovarian folliculogenesis were measured. Reproductive hormone levels in the blood were measured using a radioimmunoassay. Granulosa cells were aspirated from the ovary, and their proliferation and apoptosis were detected using immunocytochemistry. To assess secretory activity and its response to gonadotropin, ovarian fragments of treated and control ewes were cultured with and without follicle-stimulating hormone (FSH; 0, 0.1, 1, 10, or 100 IU/mL), and the release of reproductive hormones into the culture medium was evaluated. Finally, to examine the direct action of yucca on the ovary, ovarian fragments from control ewes were cultured with and without yucca extract (1, 10, or 100 μg/mL), and the release of reproductive hormones was measured. Yucca supplementation significantly decreased the size of small antral follicles (2 to <5 mm in diameter), increased accumulation of the apoptosis marker bax, and decreased serum progesterone (P4) and estradiol (E2) levels. It inhibited the release of P4 (but not other hormones), to prevent the stimulatory action of FSH on P4 output and promoted insulin-like growth factor I (IGF-I) release by fragments cultured with FSH. However, yucca supplementation did not affect the size of larger follicles and number of follicles, volume and weight of ovaries, length and weight of oviducts, caspase 3 accumulation, cell proliferation, testosterone (T) or IGF-I serum levels, or T or E2 release by cultured ovarian fragments and their response to FSH. Yucca addition to culture medium inhibited P4 and IGF-I, but not T or E2 release at the lowest (1 μg/mL) dose, and stimulated P4, but not T, E2, or IGF-I release at the highest (100 μg/mL) dose. These data suggest that yucca supplementation can reduce small antral ovarian follicle development possibly via the stimulation of apoptosis of their granulosa cells, suppression of ovarian P4 and E2 release, and alteration of ovarian IGF-I output and ovarian response to gonadotropin. Thus, yucca can directly affect P4 and IGF-I release by ovine ovarian cells.

DO PROGESTERONE, IGF-I, IGFBP-3 AND IGFBP-4 RELATE TO SEXUAL MATURATION?

Hormones and binding proteins can regulate reproduction, but their involvement in sexual maturation remains to be elucidated. This study describes possible hormonal regulators of female sexual maturation. For this purpose, the release of steroid hormone progesterone (P4), insulin-like growth factor I (IGF-I) and IGF-binding proteins (IGFBP-3, IGFBP-4) were shown in this study. Sexual maturation in gilts was found to be associated with a significant increase in the release of P4, IGF-I and IGFBP-3 in vitro. Furthermore, sexual maturation was associated with significant increase in the expression of IGFBP-3 but not in IGFBP-4. The present data obtained from in vitro study indicate that sexual maturation in females is influenced by puberty-related changes in porcine ovarian signalling substances: increase in P4, IGF-I, IGFBP-3 but not IGFBP-4. It suggests that these signalling molecules could be potential regulators of porcine sexual maturation.

Direct influence of rooibos-derived compound on rabbit ovarian functions and their response to gonadotropins

AbstractRooibos is widely used in folk medicine, but its influence on reproductive processes has been studied insufficiently. The aim of present study was to examine possible involvement of rooibos on the release of progesterone (P4), testosterone (T) and insulin-like growth factor I (IGF-I), as well as its response to gonadotropin - follicle-stimulating and luteinizing hormone (FSH+LH), the most known hormonal regulators for reproduction. Rabbit ovarian fragments were cultured in the presence of rooibos (1, 10 and 100 μg/mL medium), FSH+LH (0.01, 0.1 and 1 IU/mL medium) and rooibos (10 μg/mL medium) in combination with FSH+LH (0.01, 0.1 and 1 IU/mL medium). The release of hormones was evaluated by using radioimmunoassay. It was observed that rooibos addition to culture medium stimulated P4 and inhibited T release. IGF-I release was significantly stimulated by rooibos at all doses added. FSH+LH when given alone promoted IGF-I, but not P4 or T release. The addition of rooibos induced the suppressive effect of FSH+LH on P4 release, the stimulatory action of FSH+LH on T output and reduced the response of IGF-I to FSH+LH treatment. These data suggest that rooibos is able to directly affect the release of steroid (P4 and T) and peptide (IGF-I) hormones by rabbit ovarian cells and to modulate their response to gonadotropins.

Pulmonary Insulin-like Growth Factor I Delivery from Trehalose and Silk-Fibroin Microparticles

Insulin-like growth factor I (IGF-I) is a strong anabolic peptide with promising therapeutic value in muscle wasting diseases such as sarcopenia. We report a pulmonary IGF-I delivery system deploying silk-fibroin (SF) as carrier and in comparison to trehalose. Both IGF-I delivery systems were characterized regarding IGF-I integrity, IGF-I release profiles and aerodynamic properties. Transepithelial in vitro transport of IGF-I using the pulmonary Calu-3 model cell system followed comparable kinetics and mechanism of uptake as earlier demonstrated for insulin (INS), for which effective pulmonary delivery is known. Microparticles were spray-dried using either trehalose or SF, resulting in geometries allowing alveolar deposition. The effective IGF-I shuttling through the epithelial barrier of the lung was demonstrated in an ex vivo human lung lobe model, and expanded the exciting possibility of this administration route to this effective and anabolic peptide.

In vitro changes in secretion activity of rat ovarian fragments induced by molybdenum.

The aim of this in vitro study was to examine the secretion activity (progesterone, 17beta-estradiol and insulin-like growth factor-I) of rat ovarian fragments after molybdenum (Mo) addition. Rat ovarian fragments were incubated with ammonium molybdate (NH(4))(6)Mo(7)O(24).4H(2)O at the doses 90, 170, 330 and 500 microg.ml(-1) for 24 h and compared with control group without Mo addition. Release of progesterone (P(4)), estradiol (17beta-estradiol) and insulin-like growth factor I (IGF-I) by ovarian fragments was assessed by radioimmunoassay (RIA). Data show that P(4) release by ovarian fragments was not affected by (NH(4))(6).Mo(7)O(24).4H(2)O addition at all the doses used (90-500 microg.ml(-1)). However, addition of ammonium molybdate was found to cause a significant (P<0.05) dose-dependent decrease (at the doses 90, 170 and 500 microg.ml(-1)) in release of 17beta-estradiol by ovarian fragments in comparison to control. Also, addition of ammonium molybdate significantly (P<0.05) inhibited IGF-I release at all the doses (90-500 microg.ml(-1)) used in the study. Results suggest ammonium molybdate induced inhibition in the release of growth factor IGF-I and its dose-dependent effect on secretion of steroid hormone 17beta-estradiol but not progesterone. These data contribute to new insights regarding the mechanism of action of Mo on rat ovarian functions.

Cobalt-induced hormonal and intracellular alterations in rat ovarian fragments in vitro

The objective of this in vitro study was to examine dose-dependent changes in the secretion activity (progesterone, 17β-estradiol and insulin-like growth factor-I) of rat ovarian fragments after experimental cobalt (Co) administration including the apoptotic potential of Co on rat ovarian fragments by evaluating the expression of apoptotic markers Bax and caspase-3. Ovarian fragments were incubated with cobalt sulphate (CoSO4.7H2O) at the doses 90, 170, 330 and 500 μg.mL−1 for 24 h and compared with control group without Co addition. Release of progesterone (P4) 17β-estradiol and insulin-like growth factor-I (IGF-I) by ovarian fragments was assessed by RIA, expression of Bax and caspase-3 by SDS-PAGE and Western blotting. Observations show that P4 release by ovarian fragments was significantly (P < 0.05) inhibited after cobalt sulphate addition at higher doses 170–500 μg.mL−1 used in the study in comparison to control. However, cobalt sulphate addition did not cause any significant change in the release of 17β-estradiol by ovarian fragments at all the doses used in the study (90–500 μg.mL−1) in comparison to control. On the contrary, IGF-I release by ovarian fragments was significantly (P < 0.05) stimulated after cobalt sulphate addition at the lowest dose 90 μg.mL−1 in comparison to control, while other doses did not cause any significant change. Also, addition of cobalt sulphate decreased the expression of both the apoptotic peptides Bax and caspase-3 at the higher doses 170, 330 and 500 μg.mL−1, but not at the lowest dose 90 μg.mL−1 used in the study. Obtained results suggest Co induced (1) inhibition in secretion of steroid hormone progesterone, (2) dose-dependent increase in the release of growth factor IGF-I, and (3) decrease in the expression of markers of apoptosis (Bax and caspase-3) of rat ovarian fragments.

In vitro changes in porcine ovarian granulosa cells induced by copper

Objective of this in vitro study was to examine the secretion activity (progesterone and insulin-like growth factor I) of porcine ovarian granulosa cells after copper (Cu) addition and to outline a potential intracellular mediator (cyclin B1) of its effects. It also aimed at investigating the apoptotic potential of Cu on porcine ovarian granulosa cells after addition in vitro. Ovarian granulosa cells were incubated with copper sulphate (CuSO4·5H2O) at the doses 0.33, 0.40, 0.50, 1.0 and 2.0 μL mL−1 for 18 h and compared with control group without Cu addition. Release of progesterone (P4) and insulin-like growth factor I (IGF-I) by granulosa cells was assessed by RIA, expression of cyclin B1 by immunocytochemistry and apoptosis by TUNEL assay. Observations show that P4 release by granulosa cells was inhibited while the release of IGF-I and cyclin B1 was stimulated significantly (P < 0.05) by CuSO4·5H2O addition at the dose 2.0 μL mL−1. Also, addition of CuSO4.5H2O at the lowest dose used in the study (0.33 μL mL−1) significantly (P < 0.05) decreased apoptosis in granulosa cells. In conclusion, results indicate dose dependent effect of Cu on (1) secretion of steroid hormone progesterone and growth factor IGF-I, (2) expression of cyclin B1 as marker of proliferation of porcine ovarian granulosa cells, (3) apoptosis of porcine ovarian granulosa cells and, (4) that the effect of Cu on ovarian cell proliferation could be mediated by IGF-I and cyclin B1. Obtained data suggest interference of Cu in the pathways of proliferation of porcine ovarian granulosa cells through hormonal and intracellular peptide cyclin B1.