Androgens and estradiol-17β production by porcine uterine cells: In vitro study

Porcine (Sus scrofa domestica) uterine slices harvested during both early pregnancy and luteolysis produce steroid hormones. The aim of the present study was to determine (1) which porcine separated uterine cells secrete androgens: androstenedione (A₄) and testosterone (T), and estradiol-17β (E₂) in culture; (2) if the production of A₄, T and E₂ in the uterine cells is regulated by P4 and OT; (3) if uterine tissues expressed cytochrome P450arom gene (CYP19). Uteri were collected on Days 14 to 16 of early pregnancy and the estrous cycle. Enzymatically separated epithelial cells, stromal cells, and myocytes were cultured in vitro for 2, 6, and 12 h with control medium, progesterone (P₄; 10^-5 M), oxytocin (OT; 10^-7 M), and both hormones (P₄ + OT). The studied cells secreted A₄, T, and E₂ in vitro. Progesterone served as a substrate for steroid synthesis in the uterine cells. Isolated uterine cells, cultured separately, contributed in equal portion to the basal production of androgens (A₄ and T) during both early pregnancy and luteolysis. In pregnant pigs, the epithelial and stromal cells were rich sources of E₂ compared with myocytes. Myocytes produced E₂ mainly during luteolysis. Pregnant porcine endometrium and myometrium expressed the gene CYP19, which encodes for P450 aromatase, a steroidogenic enzyme. The results indicate an active steroidogenic pathway in porcine uterine cells. The epithelial cells, stromal cells, and myocytes participate in steroid production as an alternative source for their action in pigs.     

Preparturient changes in plasma concentrations of estrone,estradiol-17β and estradiol-17α in the fetal calf: Effects of dexamethasone infusion

The concentrations of total estrogens in fetal calf plasma were determined during a 6–10 day period immediately before delivery. Comparison was made between levels found in untreated calves and calves infused with dexamethasone at the rate of 0.1, 1.0 and 10 mg/24 hours. In untreated calves the plasma estrone, estradiol-17β and estradiol-17α levels remained relatively constant at 38 ± 7 ng ml^?1 (mean ± SEM n = 3), 46 ± 6 ng ml^?1 and 29 ± 5 ng ml^?1 respectively. Infusion with dexamethasone at 0.1 mg/24 hr (3 calves) and 1.0 mg/24 hr (3 calves) was without dramatic effect on plasma estrogen levels. However, in one fetus infused with 10.0 mg/24 hr the dexamethasone treatment may have caused a transitory rise in the levels of all estrogens examined.     

Induction of PGFM pulses and luteolysis by sequential estradiol-17β treatments in heifers

The effects of sequential induction of PGFM pulses by estradiol-17β (E2) on prominence of PGFM pulses and progesterone (P4) concentration were studied in heifers. Three treatments of vehicle (n = 12) or E2 (n = 12) at doses of 0.05 or 0.1 mg were given at 12-h intervals beginning on Day 15 postovulation. Blood samples were collected every 12 h from Days 13-24 and hourly for 12 h after the first and third treatments. On Day 15, all heifers were in preluteolysis and on Day 16 were in preluteolysis in the vehicle-treated heifers (n = 11) and either preluteolysis (n = 4) or luteolysis (n = 8) in the E2-treated heifers. Peak concentration of induced PGFM pulses during preluteolysis on Day 15 was greater (P < 0.04) than for pulses during preluteolysis on Day 16. The interval from ovulation to the beginning of luteolysis was shorter (P < 0.04) in the E2-treated heifers than in the vehicle-treated heifers. An E2-induced PGFM pulse was less prominent (P < 0.008) in heifers in temporal association with a transient resurgence in P4 than in heifers with a progressive P4 decrease. The hypothesis that repeated E2 exposure stimulates increasing prominence of PGFM pulses was not supported. Instead, repeated exposure reduced the prominence of PGFM pulses, in contrast to the stimulation from the first E2 treatment. Reduced prominence of a PGF_2α pulse during luteolysis can lead to a transient resurgence in P4 concentration.     

Interconversion of estrone and estradiol-17β in lung slices of the adult human

The metabolism of tritium-labeled estrone and estradiol-17β in slices of lung tissue obtained from an adult human was studied: estrone was identified as the only metabolite of estradiol-17β and estradiol-17β as the exclusive product of estrone metabolism. Product formation remained linear as a function of time of incubation up to 3 h and of wet lung tissue mass up to 300 mg/ml. At equimolar substrate concentrations, the rates of estrone formation were at least 2-fold greater than those of estradiol-17β. The apparent K_M of 17β-hydroxysteroid oxidoreductase for estrone was 11 μM and that for estradiol-17β was 10 μM. These results are suggestive that the human lung enzyme binds estrone and estradiol-17β with similar affinities; however, the oxidative pathway is favored as indicated by the greater V_max attained in the formation of estrone. It is possible that, in vivo, the human lung constitutes a site for estradiol-17β inactivation to estrone as well as a site for the conversion of estrone to estradiol-17β. This last process may become particularly important in instances in which the ovaries have ceased to function and secrete estradiol-17β, e.g. the postmenopausal women.     

Autoradiographic study on the localization of estradiol-17β in neonatal mouse uterus and cervix

Summary The uptake of ³H-estradiol-17 in the neonatal mouse uterus and cervix has been studied by an autoradiographic method. When the radio-active hormone is administered in vivo and in vitro, grains are found to be concentrated above the nuclei both in the uterine and cervical epithelium and stroma. Grain counts revealed that the nuclear concentration of grains is higher at 4 h than at 2 h after isotope injection. The cervical epithelium has a higher nuclear concentration than the uterine epithelium both in vivo and in vitro. In the stroma, this situation is reversed except after in vitro treatment of the tissues.In the cervix, more of the hormone seems to be located within the nucleus while in the uterus a higher proportion of the grains are found in the vicinity of the nuclear periphery.Although the nuclear concentration of grains is higher at 4 h than at 2 h, the number of grains above the sections is lower at 4 h. Both in vivo and in vitro, the number of grains is higher above the stromal than above the epithelial compartments of the uterus and cervix.Five days old animals showed the same labeling pattern. The differences in uptake and distribution of ³H-estradiol are discussed in relation to other known differences in the hormone responsiveness in these tissues.We are greatly indebted to Professor W.E. Stumpf and the Laboratories for Reproductive Biology, University of North Carolina Medical School for the opportunity to study the method of dry mount autoradiography. The work has been supported by the Norwegian Research Council for Science and the Humanities and by the Norwegian Cancer Society (Landsforeningen mot Kreft)     

Differential regulation of estrous behavior and luteinizing hormone secretion by estradiol-17β in ovariectomized dairy cows

Holstein cows (n = 9) were used in an experiment to characterize the behavioral and endocrine responses to estradiol-17β when administered at rates designed to maintain peripheral concentrations within a physiological range. Cows were pretreated with progesterone for 3 d. Three days after progesterone treatment was completed, each cow was assigned to one of five estradiol-17β treatment groups (Doses 0 to 4), calculated to produce and maintain 0, 3, 6, 9, or 12 pg/mL in peripheral blood for 8 h. The experiment was conducted in eight replicates (with 3 to 7 cows each), with no dose repeated in any replicate. In each replicate, at least one additional cow was given an injection of estradiol-17β (500 μg im, in a corn oil vehicle) to facilitate estrus detection. Estrus was detected by visual observation for 30 min at 4 h intervals. Estrus was defined as a cow that stood to be mounted at least twice during the 50 h interval over which estrus was observed. Jugular venous blood samples were collected at 2 h intervals throughout the infusion and observation periods for quantification of luteinizing hormone (LH). Cows that received the highest dose (Dose 4, n = 7) all showed estrus, whereas those that received the two lowest doses (Dose 0, n = 5; Dose 1, n = 6) did not. Over the course of the experiment, five cows received each dose at least once. Of these, three showed estrus at Doses 2, 3, and 4, whereas the other two showed estrus only at Dose 4. Therefore, individual cows differed in the amount of estradiol-17β needed to induce estrus. There was a linear effect of dose on duration of estrus (P < 0.01). Estrus was shorter for Dose 2 (8.0 h) than for Dose 4 (18.4 h). The onset of estrus (after start of infusion) tended to be later for Dose 2 (20.0 h) than for Doses 3 and 4 (14.0 and 13.4 h, respectively; P = 0.15). Preovulatory-like surges of LH were induced in all cows at Doses 2, 3, and 4. Surges also were detected in 3 of 5 cows receiving Dose 1. The magnitude of the LH surge was less for Doses 1, 2, and 3 than for Dose 4 (P = 0.06). In contrast to the timing of estrus, the timing of the LH surge (after start of infusion) was not different among doses (P = 0.88). Thus, the hypothalamic centers responsible for regulating expression of estrus and secretion of LH responded differently to estradiol-17β.     

Production,clearance rates and metabolic fate of estradiol-17β in the plasma of the laying hen

A single dose of tritiated estradiol-17β (³H-E₂β) was injected i.v. into 5 high egg producing White Leghorn hens, 31 weeks of age, at 19.2 ± 2.1 (mean ± S.D.) hr before oviposition. Blood (2 ml) was sampled at approximately 5 min intervals over 40 min. Whenever possible, metabolites were monitored and identified by the double isotope technique with the addition of the corresponding ¹⁴C-labelled standards to plasma prior to analysis. The metabolic half-life and clearance rate of ³H-E₂β in plasma were 10.9 ± 1.9 min and 118 ± 18 ml/min/kg body weight, respectively. The calculated production rate of E₂β at 19.2 hr before oviposition was 19.5 ± 5.7 ng/min based on the plasma level (93±22 pg/ml) measured at that time. The relative concentrations (% of plasma radioactivity) of the major metabolites isolated at 5.7 ± 0.6 min post injection were, in descending order: estradiol-17β-3-sulfate (E₂β-3S : 14.9 ± 2.7), estradiol-17α-3-sulfate (E₂α-3S; 5.7 ± 0.3), estrone (E₁; 4.6 ± 0.5), estrone sulfate (E₁S; 2.2 ± 0.5), and estradiol-17 α (E₂α; 1.2 ± 0.4). As time proceeded, the relative concentration of E₂α-3S gradually increased so that by 43.2 ± 1.0 min it became the most abundant identifiable metabolite (12.3 ± 1.1) followed by E₂β-3S (9.1 ± 1.7), E₂S (1.2 ± 0.6), E₁ (0.7 ± 0.4) and E₂α (0.3 ± 0.2). These findings are consistent with the view that one of the major pathways of E₂β metabolism in the circulation of the hen is via E₂β

E₂β?3S ?E₁S

E₂α-3S.     

Effects of estradiol-17β in the male xenopus laevis: Isolation and translation of cytoplasmic messenger rna populations

Summary Total Xenopus liver cytoplasmic RNA isolated following long-term estrogen administration (14 days) was fractioned using Sepharose 4B chromatography. One of the Sepharose 4B peaks was shown to contain RNA with a molecular weight reported for vitellogenin mRNA (34S). The presence of estrogeninduced vitellogenin mRNA in the peak 5 RNA was determined by translation of the RNA in the oocyte and analysis of the oocyte translational products by immunoprecipitation with anti-vitellogenin.Sepharose 4B peaks 2 and 3 were also observed to contain estrogen induced mRNA populations sedimenting between 9-18S. These findings suggest that Sepharose 4B chromatography might prove useful in separating different mRNA populations following estrogen-induced gene activation.     

Short-term bioaccumulation, circulation and metabolism of estradiol-17β in the oyster Crassostrea gigas

Steroids are active signal transmitters in Vertebrates. These roles have also been hypothesized in other Phyla and endocrine disrupting effects have been reported for different estrogen-like compounds in fishes and some marine invertebrates. As estradiol-17β has shown some physiological activities in the oyster and as estrogens or estrogen-like molecules can be present in water, we have investigated the bioaccumulation and metabolism of this estrogen in vivo in the oyster Crassostrea gigas. When dissolved in seawater, in less than 48 h estradiol-17β concentrated up to 31 times in the soft tissues of the suspension-feeder mollusc. Injected in the adductor muscle, estradiol-17β circulated from muscle to the gonad, the gills, the mantle, the labial palps, and to a lesser extent to the digestive gland. After 2 h, estradiol flow increased specifically towards this gland. Different hypotheses were raised concerning the circulation paths. However, in all cases estradiol metabolism primarily evidenced an in vivo transformation into estrone in the whole oyster and in its digestive gland. This strong 17β-hydroxysteroid-dehydrogenase activity confirms our previous in vitro results. In conclusion, it is proposed that oyster is able to take in charge estradiol as a potential contaminant in seawater. Therefore, its bioaccumulation and transformation into estrone could be studied as potential biomarkers of endocrine disruption. Furthermore, the experimental approach with dissolved steroids in the seawater combined to an anatomical screening appears as an interesting tool to investigate the bivalve endocrinology.     

Effects of estradiol-17β on cholesterol metabolism in the rat: A study using a deuterium label and mass spectrometry

The effect of estradiol-17β (E₂) on several important aspects of cholesterol metabolism were examined in the rat. Ovariectomized rats were implanted subcutaneously with 1 or 4 cm. of silastic tubing packed with E2, and were also given 2% D₂O in their drinking water. The E2 diffused slowly out of the implants and the two different lengths of tubing resulted in constant E2 blood concentrations of either high (4.0 cm) or physiological (1.0 cm) levels. By measuring the rate of incorporation of deuterium into plasma cholesterol by mass spectrometry over a period of 42 days, we determined the rate constant of cholesterol synthesis and cholesterol turnover time and rate under two E2 dosage conditions. E2 treatment did not affect the rate constant of cholesterol synthesis or the cholesterol turn-over time. However, cholesterol turnover rate (mg synthesized/day) showed a dose dependent reduction with increasing doses of E2. This may be secondarily caused by E2's suppression of both food Intake and subsequent weight gain; E2 treated animals are smaller and, therefore, synthesize less cholesterol per day. Additionally, E2 treated animals showed a rise in plasma cholesterol levels and in the fraction of labeled cholesterol appearing in the plasma.     

Chronic Sleep Restriction Induces Aβ Accumulation by Disrupting the Balance of Aβ Production and Clearance in Rats

Amyloid-β (Aβ) plays an important role in Alzheimer’s disease (AD) pathogenesis, and growing evidence has shown that poor sleep quality is one of the risk factors for AD, but the mechanisms of sleep deprivation leading to AD have still not been fully demonstrated. In the present study, we used wild-type (WT) rats to determine the effects of chronic sleep restriction (CSR) on Aβ accumulation. We found that CSR-21d rats had learning and memory functional decline in the Morris water maze (MWM) test. Meanwhile, Aβ₄₂ deposition in the hippocampus and the prefrontal cortex was high after a 21-day sleep restriction. Moreover, compared with the control rats, CSR rats had increased expression of β-site APP-cleaving enzyme 1 (BACE1) and sAPPβ and decreased sAPPα levels in both the hippocampus and the prefrontal cortex, and the BACE1 level was positively correlated with the Aβ₄₂ level. Additionally, in CSR-21d rats, low-density lipoprotein receptor-related protein 1 (LRP-1) levels were low, while receptor of advanced glycation end products (RAGE) levels were high in the hippocampus and the prefrontal cortex, and these transporters were significantly correlated with Aβ₄₂ levels. In addition, CSR-21d rats had decreased plasma Aβ₄₂ levels and soluble LRP1 (sLRP1) levels compared with the control rats. Altogether, this study demonstrated that 21 days of CSR could lead to brain Aβ accumulation in WT rats. The underlying mechanisms may be related to increased Aβ production via upregulation of the BACE1 pathway and disrupted Aβ clearance affecting brain and peripheral Aβ transport.

     

Effect of 17α-methyltestosterone,estradiol-17β and synthetic LHRH on production of gonadotropic hormone in pituitaries of rainbow trout (organ culture)

Pituitary glands from 6-month-old sexually immature female rainbow trout, Salmo gairdneri, were kept in organ culture for 48 or 72 h. Certain groups of pituitaries were cultivated for 48 h on either control medium or medium with 17 alpha-methyltestosterone (MT), or with estradiol-17 beta (E2) in concentrations of 8.5 X 10(-7) M. Other groups of pituitaries were cultivated for 72 h on control medium, or for 48 h on either control medium or MT-medium or E2-medium, and subsequently for 24 h on medium with synthetic LHRH in concentrations of 8.5 X 10(-7) M and 8.5 X 10(-10) M. Gonadotropic (GTH) cells are identified by Alcian Blue-Periodic Acid Schiff-Orange G staining and the double-antibody immunoenzyme-cytochemical technique using anti-carp beta GTH as the first antibody. A quantitative histological procedure was used to study the nuclear size of the GTH cells in response to the different hormones. Secretory activity was estimated by measuring the gonadotropin (GTH) content in extracts of pituitaries, plasma, and the culture media every 24 h by radioimmunoassay. Cultivation on MT- or E2-enriched medium results in an increase of the total amount of GTH in the pituitary and medium, an accumulation of GTH in GTH-cells (approximately 20 percentage points) and an increase in their nuclear size, indicating a stimulation of GTH synthesis. However, autonomous GTH-release is not affected by these steroids. Subsequent cultivation of the pituitaries for 24 h with LHRH causes stimulation of GTH synthesis (approximately 20 percentage points). Preincubation with steroids increases the GTH synthesis capacity of LHRH only when used in a concentration of 8.5 X 10(-10) M. Moreover, 8.5 X 10(-7) M LHRH causes a stimulation of GTH-release. Preincubation of the pituitaries with steroids increases the responsiveness of GTH-cells to LHRH. It is concluded that GTH-production in pituitaries of immature female rainbow trout can be directly influenced by gonadal steroids and by a hypophysiotropic substance.     

Serum chorionic gonadotropin,progesterone, and estradiol-17β levels during pregnancy in the common marmoset,Callithrix jacchus

The serum levels of chorionic gonadotropin (LH/CG), progesterone, and estradiol-17β were measured during pregnancy in the common marmoset. The gestation period in five females was 144±1.5 (141–145) days. The LH/CG level increased from the early stage of pregnancy, reached a maximum of 10–17 ng/ml at 50 to 70 days and decreased to under 40 pg/ml at about 100 days. The progesterone level maintained the same value as that at the luteal phase of 20–40 ng/ml until 90 days of pregnancy, when the serum LH/CG was declining, thereafter increased abruptly, reached a maximum of 140–210 ng/ml at 110–130 days and fell to a low level of under 0.4 ng/ml at 5–10 days before parturition. The estradiol-17β was less than 2 ng/ml until 90 days of pregnancy, thereafter increased abruptly and maintained a high level of 40–135 ng/ml until just before parturition. The 3β-hydroxysteroid dehydrogenase activity in the placenta of the common marmoset was 40 times higher than that in the fetal adrenal, while in the Japanese monkey the former was only about one 40th of the latter. The time course of the serum progesterone and estradiol-17β during pregnancy and the role of the placenta which synthesized and secreted these hormones in the common marmoset showed a similarity to those of humans and anthropoid apes rather than those ofMacaca species. The common marmoset represents a valuable animal model for investigating the feto-placental unit in humans.     

Correlation of plasma estradiol-17β and progesterone levels with ultrastructure and histochemistry of ovarian follicles in the white-spotted char,Salvelinus leucomaenis

Summary Plasma estradiol-17 and progesterone profiles were correlated with morphological changes in ovarian follicles during the preovulatory and postovulatory periods in the white-spotted char, Salvelinus leucomaenis. Plasma estradiol levels were highest in September, and were followed by a sharp drop in October; they remained very low throughout the postovulatory period. There was a good correlation between plasma estradiol levels and the gonadosomatic index, thus suggesting that estradiol is responsible for the synthesis of vitellogenic proteins. Plasma progesterone levels were very low in August, began to rise in September and reached a peak soon after ovulation; progesterone remained high for several days after ovulation. A preovulatory rise in plasma progesterone levels was recorded, and this is discussed in relation to the induction of oocyte maturation.In the preovulatory follicles, neither granulosa cells nor special thecal cells (ST cells) showed ⁵, 3-hydroxysteroid dehydrogenase (3-HSD) activity. In the young postovulatory follicles, in contrast, the ST cells showed intense 3-HSD activity with extensive agranular endoplasmic reticulum and numerous large mitochondria, while granulosa cells did not show 3-HSD activity. These results strongly suggest that the ST cells are the major sites of progesterone synthesis during the postovulatory period.Nanae Fish Culture Experimental Station Contribution No 14     

Effects of dietary estradiol-17β on feminization,growth and body composition in the Japanese eel (Anguilla japonica)

• 1.1.Juvenile Japanese eels (Anguilla japonica) were fed on a diet supplemented with estradiol-17β (E₂) at doses of 25, 50 and 75 mg/kg. The effects on growth, sex distribution and body composition were investigated in two groups of gonadally undifferentiated stages (early and later juvenile stages).
• 2.2.Feminization (95–100%) was observed in all E₂-treated groups.
• 3.3.The growth rate of fish treated with 25 and 50 mg/kg E₂ diet at the early juvenile stage was significantly increased.
• 4.4.The amount of protein in muscle decreased and that of fat increased in the E₂-treated groups except in the early juvenile stage fed with 25 mg/kg E₂.

     

The induction of male sexual behavior in red deer (Cervus elaphus) by the administration of testosterone to hinds and estradiol-17β to stags

One-gram implants of testosterone were placed subcutaneously in two adult intact and one castrated red deer hinds, and 100-mg implants of estradiol-17β were given to three castrated and two intact red deer stags. Their sexual behavior was then observed in the wild and in captivity for up to 3 years. Testosterone initially produced prolonged and intense estrous behavior in the hinds; this gradually gave way to behavior normally shown only by rutting stags, such as flehmen, herding threats, and roaring. Testosterone-implanted hinds rose in the dominance hierarchy as measured by competition for food. Pregnancy, or the administration of progestagens, suppressed most of this testosterone-induced behavior. Testosterone also induced development of male secondary sexual characteristics such as formation of small antler pedicles (but not antlers), hypertrophy of the clitoris and neck musculature, and development of a neck mane and male rutting odor. Estradiol-17β mimicked the behavioral effects of testosterone when given to castrated stags, stimulating all components of rutting behavior. In intact stags, the only effects were to abolish antler casting and stimulate roaring; normal rutting behavior continued unchanged. In contrast to the effects of testosterone, estradiol-17β did not influence the social status of either intact or castrated stags. The distinctions between the behavioral effects of androgens and estrogens are ill defined and are determined more by the prior sexual differentiation of the brain and the duration of steroid treatment, than by the nature of the steroid.     

Serum LH,progesterone and estradiol-17β levels throughout the ovarian cycle,during the early stage of pregnancy and after the parturition and the abortion in the common marmoset,Callithrix jacchus

In the ovarian cycle of common marmosets, serum progesterone began to increase at two to three days after estradiol-17β or LH surge, attained a peak of 25–70 ng/ml and then declined to a level of under 2 ng/ml before the ensuing rise in estradiol-17β and LH. Serum estradiol-17β increased to 700–5,500 pg/ml during the luteal phase, synchronizing with progesterone. It is suggested that the corpus luteum secreted estradiol-17β as well as progesterone. The cycle length as determined from the interval between successive LH surges was approximately 28 days. During the luteal phase, the levels of progesterone and estradiol-17β were higher than in Old World monkeys and women, but marmosets were not accompanied by any clinical symptoms due to excessive progesterone and estradiol-17β. This suggests that such unresponsiveness to progesterone and estradiol-17β in marmosets reflects the small amount of estradiol-17β receptor and presumably also the lower function of the post receptor system. Recovery of the post-partum ovarian cycle in two marmosets differed from that observed in Old World monkeys and women. The first LH surge was found on the ninth and tenth day after parturition and the first ovulation led to the next pregnancy. This suggests that the suckling stimulus of newborns in the common marmoset does not cause any delay in recovery of the ovarian cycle. In three cases of abortion, the recovery of the ovarian cycle was almost the same as that in the case of normal parturition: the first LH surge appeared on the 10th, 14th, and 34th day after abortion.     

Activation of the PI3K pathway increases TLR-induced TNF-α and IL-6 but reduces IL-1β production in mast cells

Recognition of bacterial constituents by mast cells (MCs) is dependent on the presence of pattern recognition receptors, such as Toll-like receptors (TLRs). The final cellular response, however, depends on the influence of multiple environmental factors. In the current study we tested the hypothesis that the PI3K-activating ligands insulin-like growth factor-1 (IGF-1), insulin, antigen, and Steel Factor (SF) are able to modulate the TLR4-mediated production of proinflammatory cytokines in murine MCs. Costimulation with any of these ligands caused increased LPS-triggered secretion of IL-6 and TNF-α, but attenuated the production of IL-1β, though all three cytokines were produced in an NFκB-dependent manner. The pan-specific PI3K-inhibitor Wortmannin reverted the altered production of these cytokines. In agreement, MCs deficient for SHIP1, a negative regulator of the PI3K pathway, showed augmented secretion of IL-6/TNF-α and reduced production of IL-1β in response to LPS alone. The differential effects of IGF-1 on TLR4-mediated cytokine production were also observed in the context of TLR2 and IL-33 receptor-mediated MC activation. Importantly, these effects were seen in both bone marrow-derived and peritoneal MCs, suggesting general relevance for MCs. Using pharmacological and genetic tools, we could show that the p110δ isoform of PI3K is strongly implicated in SF-triggered suppression of LPS-induced IL-1β production. Costimulation with antigen was affected to a lesser extent. In conclusion, NFκB-dependent production of proinflammatory cytokines in MCs is differentially controlled by PI3K-activating ligand/receptor systems.