Insulin-like Growth Factor-1 mRNA Expression Research Articles

Maternal nutritional restriction during late gestation impairs development of the reproductive organs in both male and female lambs

Maternal nutritional restrictions during late gestation could lead to fetal hypoglycemia. Glucose levels in the fetal sheep regulate circulating insulin-like growth factor 1 (IGF1) levels, which stimulate cell proliferation and differentiation of reproductive organs after binding to its own receptor or estrogen receptors. The objective of this study was to determine the effects of subnutrition of ewes during the last trimester of gestation on the serum glucose/IGF1 levels and development of reproductive organs in their lambs.Pregnant ewes carrying singletons were randomly assigned to restricted (R ewes, n = 8) or control (C ewes, n = 8) groups (4 lambs of each gender/group) and fed with 50% or 100% of metabolic energy requirements from ∼100 days of gestation to term (∼147 days), respectively. Blood samples from lambs were taken on the first day after born and once at week for serum glucose and IGF1 determination. Lambs were euthanatized at 2 months of age, reproductive organs were weighted and tissue samples were collected from them for histology and to measure mRNA expression of IGF1 and its receptor (IGF1R) by qRT-PCR. Pre-partum glucose levels in R ewes were significantly lower compared to C ewes (p < .05). Compared to lambs born from C ewes, lambs born from R ewes showed lower serum levels of glucose and IGF1 during the first week of age (p < .05). At 2 month of age, these lambs had significant lower uterine and testicular weight and lower ovarian, uterine and testicular mRNA expressions of IGF1 and its receptor (p < .05). Histological findings showed that diameter of secondary and tertiary follicles in ovaries and number of endometrial glands in uterus of females, or number of Sertoli cells and seminiferous tubules and diameter, perimeter and tubular area in testicles of males were significantly lower in lambs born from R ewes compared to the respective organs of lambs born from the C ewes (p < .05). In conclusion, these results demonstrate that maternal subnutrition during late gestation affects IGF1 levels during fetal life and impairs reproductive development in the neonatal lamb, which could have permanent negative consequences in the future reproductive performance of the offspring.

Efficacy of topical tofacitinib in promoting hair growth in non-scarring alopecia: possible mechanism via VEGF induction.

Tofacitinib is a Janus kinase 3 (JAK3) inhibitor that promotes hair growth; however, the efficacy and mechanism of this effect are not yet understood. This study aimed to evaluate the efficacy and mechanism of topical tofacitinib on hair growth in mice. Eight-week-old male C57BL/6 mice were divided equally into four groups and treated topically with tofacitinib, minoxidil, or vehicle once daily for 21days. Weekly photographs were taken to determine the area and rate of hair growth, and tissue samples were collected for histopathological evaluation. mRNA and protein expression of anagen-maintaining growth factors, including vascular endothelial growth factor (VEGF) and insulin-like growth factor-1 (IGF-1), were determined via RT-PCR and ELISA, respectively. Tofacitinib-treated mice exhibited more hair regrowth than either minoxidil-treated or control mice did between day 7 and 21 (P<0.05). Topical tofacitinib also promoted more rapid hair growth rate than topical minoxidil or control did (P<0.001). Histopathology showed a distinct increase in the number of hair follicles, mostly in the anagen phase, in the tofacitinib-treated group. Hair follicles in the minoxidil- and vehicle-treated groups were more often classified as catagen and anagen. VEGF mRNA and protein expression in the tofacitinib-treated group was significantly greater than those in the other groups (P<0.05). IGF-1 mRNA expression was not upregulated in tofacitinib-treated mice. Topical tofacitinib is effective in promoting hair growth, and the possible mechanism involves increased VEGF levels and lowered inflammation. This study will help develop a new therapeutic option for non-scarring alopecia.

Central and peripheral administrations of insulin-like growth factor-1 suppress food intake in chicks

A number of studies have been made on the physiological actions of insulin-like growth factor-1 (IGF-1) in mammals and birds. In mammals, the effects of central administration of IGF-1 on food intake have been examined. For example, intracerebroventricular administration of IGF-1 significantly decreased food intake in diabetic rats, but not in sheep and nondiabetic rats. The chicken is known to be a hyperglycemic animal. Like satiety hormones, plasma IGF-1 levels are elevated postprandially in chickens. In this study, we hypothesized that IGF-1 is involved in the regulation of food intake in chickens. Intracerebroventricular administration of IGF-1 significantly suppressed food intake in chicks in a dose dependent manner. Both the mRNAs of IGF-1 and its receptor were expressed throughout the brain. However, the mRNA levels of IGF-1 were not influenced by fasting and refeeding in all regions of the brain. On the other hand, 6h of fasting significantly suppressed mRNA expression of hepatic IGF-1, and this effect was significantly reversed by 6h of refeeding. Furthermore, intravascular administration of IGF-1 significantly suppressed food intake in chicks. These findings suggest that IGF-1 may function as a satiety hormone in chickens.

Maternal betaine supplementation in rats induces intergenerational changes in hepatic IGF-1 expression and DNA methylation.

Betaine is widely used in animal nutrition to promote growth. Here, we aimed to investigate whether maternal betaine supplementation during pregnancy can exert multigenerational effects on growth across two generations and the possible epigenetic modifications associated to such effects. In this study, 3-month-old female Sprague-Dawley rats were fed diet supplemented with 1% betaine throughout the pregnancy and lactation. Betaine-supplemented dams produced bigger litter but smaller F1 pups at birth and weaning. However, F2 pubs had higher weaning weight. In accordance with the growth performance, serum insulin-like growth factor 1 (IGF-1) levels were significantly lower in F1 yet higher in F2 pups, so was hepatic IGF-1 mRNA expression. Concurrently, dietary betaine supplementation to F0 dams increased hepatic expression of betaine homocysteine methyltransferase, at both mRNA and protein levels, in F1, but not F2 pups. Moreover, hepatic IGF-1 gene promoter 1 was detected to be significantly hypermethylated in F1 pups, whereas both promoters 1 and 2, together with almost all exons, were found to be hypomethylated in F2 offspring. Maternal betaine supplementation during pregnancy and lactation exerts distinct effects on growth of F1 and F2 rat offspring, probably through differential modification of IGF-1 gene methylation and expression in liver.

Dietary protein-induced hepatic IGF-1 secretion mediated by PPARγ activation.

Dietary protein or amino acid (AA) is a crucial nutritional factor to regulate hepatic insulin-like growth factor-1 (IGF-1) expression and secretion. However, the underlying intracellular mechanism by which dietary protein or AA induces IGF-1 expression remains unknown. We compared the IGF-1 gene expression and plasma IGF-1 level of pigs fed with normal crude protein (CP, 20%) and low-protein levels (LP, 14%). RNA sequencing (RNA-seq) was performed to detect transcript expression in the liver in response to dietary protein. The results showed that serum concentrations and mRNA levels of IGF-1 in the liver were higher in the CP group than in the LP group. RNA-seq analysis identified a total of 1319 differentially expressed transcripts (667 upregulated and 652 downregulated), among which the terms “oxidative phosphorylation”, “ribosome”, “gap junction”, “PPAR signaling pathway”, and “focal adhesion” were enriched. In addition, the porcine primary hepatocyte and HepG2 cell models also demonstrated that the mRNA and protein levels of IGF-1 and PPARγ increased with the increasing AA concentration in the culture. The PPARγ activator troglitazone increased IGF-1 gene expression and secretion in a dose dependent manner. Furthermore, inhibition of PPARγ effectively reversed the effects of the high AA concentration on the mRNA expression of IGF-1 and IGFBP-1 in HepG2 cells. Moreover, the protein levels of IGF-1 and PPARγ, as well as the phosphorylation of mTOR, significantly increased in HepG2 cells under high AA concentrations. mTOR phosphorylation can be decreased by the mTOR antagonist, rapamycin. The immunoprecipitation results also showed that high AA concentrations significantly increased the interaction of mTOR and PPARγ. In summary, PPARγ plays an important role in the regulation of IGF-1 secretion and gene expression in response to dietary protein.

Heat exposure alters the mRNA expression of growth- and stress-related genes in chicks

High ambient temperature, a major stressor, impairs the growth of chickens. In this study, we examined the effect of heat exposure on the mRNA expression of various growth related genes, such as growth hormone (GH), insulin-like growth factor-1 (IGF1), GH-releasing hormone (GHRH), thyrotropin-releasing hormone (TRH), and somatostatin (SST) in layer-type chickens (Gallus gallus domesticus) at an early age. In addition, we examined the reactivity of the corticotrophin-releasing hormone (CRH) system, one of the stress-regulating pathways, to heat exposure and its role in altering the growth related genes. Four-day heat exposure reduced the body weight gain, feed intake, and feeding efficiency and increased the rectal temperature of chicks (P<0.05). The mRNA expression levels of pituitary GH, liver IGF1, and diencephalic GHRH decreased with heat exposure (P<0.05) whereas the levels of TRH or SST did not change. Heat exposure also reduced the diencephalic mRNA expression level of CRH and increased pituitary CRH receptor-2 mRNA and the plasma corticosterone (CORT) concentration (P<0.05), suggesting that heat exposure affected the hypothamic-pituitary adrenal gland (HPA) axis. Similar to the heat-exposure study, subcutaneous injection of CORT for 4 days decreased body weight, and the mRNA expression of pituitary GH and liver IGF1 (P<0.05). The present study demonstrated that heat exposure reduced the mRNA expressions of pituitary GH and liver IGF1, and suggested that the change in mRNA expression may have been partly caused by CORT.

Impacts on thepregnancy outcome in the mice of controlled ovarian hyperstimulation treated with acupuncture at different time points

To explore the optimal time of acupuncture intervention in the assisted reproduction. One hundred and twenty female mice and 60 male mice were collected. 20 female mice were selected in the natural period group and the rest 100 female mice were prepared as the model of controlled ovarian hyperstimulation (COH). The model mice were randomized into a COH group, a down-regulation group, a gonadotropins (Gn) start group, an injection of human chorionic gonadotropin (HCG) group and an embryo culture group, 20 mice in each one. The donor mice and receptor mice were subdivided in each group, 10 mice in each subgroup. One week before the experiment, vas deferens ligature was done in 30 male mice and the other 30 male mice did not receive ligature. In the down-regulation group, the Gn start group, the HCG injection group and the embryo culture group, electroacupuncture (EA) was applied to "Guanyuan" (CV 4), "Zhongji" (CV 3) and "Sanyinjiao" (SP 6) at the time points accordingly. EA stimulation was in the condition of continuous wave, 2 Hz and 1 mA. No inter-vention was given in the natural period group and the COH group. On the day of HCG injection, the donor mice and the non-ligatured male mice were put in the same cage of each group. The fertilized ovum was collected with the date of fertilization marked and was fostered in the incubator. At the ratio of 1:1, the receptor mice and ligatured mice were put in the same cage in each group. The vaginal plug was examined in the next morning. The pseudopregnancy was marked with the date of plug observed. In the 68th hour of embryo culture, the embryo of the donor was shifted to the receptor on the same day when the plug was observed. The clinical pregnancy rate and embryo imbed site number were observed. RT-PCR assay was adopted to determine the expression of insulin-like growth factor-1 (IGF-1) mRNA in endometrium. In the COH group, the pregnancy rate, average imbed site number and endometrial IGF-1 mRNA expression were all significantly lower than those in the natural period group (all P<0.01). After EA treatment, in the Gn group, the HCG injection group and the embryo culture group, the pregnancy rates were higher significantly than those in the COH group (P<0.05, P<0.01). In the HCG injection group, the average imbed site number and IGF-1 mRNA expression were increased apparently as compared with those in the COH group (both P<0.01), better than those in the Gn group and the embryo culture group (all P<0.01). In the treatment with acupuncture combined with IVF-ET for infertility, the intervention of acupuncture on the day of HCG injection is the optimal time point. It increases the secretion of endometrial IGF-1 so as to improve the clinical pregnancy rate, the mean imbed site number and the embryo implantation.

Effects of Dietary Supplementation with Different Polyunsaturated Fatty Acids on Expression of Genes Related to Somatotropic Axis Function in the Liver, Selected Blood Indicators, Milk Yield and Milk Fatty Acids Profile in Dairy Cows

Abstract The objective of this study was to investigate whether dietary supplementation with different polyunsaturated fatty acids (PUFA s) affects expression of genes related to somatotropic axis and the plasma concentrations of insulin, glucose, non-esterified fatty acids (NEFA), beta hydroxyl butyrate acids (BHBA) and insulin-like growth factor 1 (IGF1) and milk fatty acids profile. Right after calving, Holstein cows (n=45) were randomly assigned to one of three diets supplemented with roasted whole soybean as a source of omega-6 PUFA (omega-6, n=15), linseed as a source of omega-3 PUFA (omega-3, n=15) or palm oil (control, n=15). Each cow was in the study over a period of 70 days. Blood samples were collected every two weeks from day 1 to 70 of lactation and plasma concentrations of insulin, glucose, NEFA, BHBA and IGF1 were determined. Liver samples were taken from a subset of 18 cows (6 per diet) at day 70 postpartum and hepatic mRNA level of total growth hormone-receptor 1A (GHR1A), insulin receptor (INSR), IGF1 and insulinlike growth factor binding protein (IGFBP2) was assessed. Experimental diets did not affect milk yield. Plasma glucose and insulin concentrations were greater for omega-3 treatment compared to omega-6 and control treatments. Cows fed diets enriched in omega-3 exhibited greater INSR and GHR1A mRNA expression, and a tendency for greater IGF1 mRNA expression in the liver compared to omega-6 and control cows. Plasma IGF1 concentration was significantly higher in omega-3 treatment compared with omega-6 and control treatments. Results of this study suggest that feeding omega-3 PUFA s during early postpartum couples with the somatotropic axis, leading to an increase in plasma IGF1 concentration in dairy cows.

EXPERIMENTAL STUDY ON LENTIVIRUS-MEDIATED MULTI-GENES CO-TRANSFECTION IN BONE MARROW MESENCHYMAL STEM CELLS FOR TREATMENT OF KNEE OSTEOARTHRITIS IN CYNOMOLGUS MONKEY

To observe the effect of lentivirus-mediated cyclooxygenase 2 (COX-2) and Aggrecanase-1 silencing and insulin-like growth factor 1 (IGF-1) in BMSCs after injecting into the knee joint cavity in cynomolgus monkeys with knee osteoarthritis (OA). BMSCs were isolated from the bone marrow of 10 donors. The lentivirus vector expressing genes of COX-2, Aggrecanase-1, and IGF-1 were constructed, and transfected into the third generation human BMSCs at 40 multiplicity of infection (virus group); BMSCs transfected with lentivirus-empty vector served as blank-virus group. The growth status and number of BMSCs were observed under inverted phase contrast microscope, and normal BMSCs were used as normal control group. At 1 week after transfected, the mRNA expressions of COX-2, Aggrecanase-1, and IGF-1 were detected with RT-PCR. Nine 3-year-old cynomolgus monkeys were selected to establish the OA model according to Hulth modeling method, and were randomly divided into 3 groups (n=3). At 6 weeks after remodeling, the right knee joint cavity was injected accordingly with 1 mL BMSCs (about 1×107 cells) in virus group and blank-virus group, with 1 mL of normal saline in the blank control group; the left knee served as normal controls. The general condition was observed after injection; at 1, 4, and 6 weeks, the concentrations of prostaglandin E2 (PGE2), IL-1, Aggrecanase-1, and IGF-1 of double knee liquid were detected with ELISA; at 6 weeks, MRI, general observation, histology method, and immunohistochemistry method were used to detect the knee cartilage changes and the expressions of COX-2, Aggrecanase-1, and IGF-1 were measured with RT-PCR. No significant difference was found in cell morphology and growth curve between 2 groups after transfection. By RT-PCR, COX-2, and Aggrecanase-1 expressions were significantly reduced, IGF-1 expression was significantly increased in virus group when compared with normal control group and the blank-virus group (P<0.05). All monkeys survived to the end of the experiment after injection. When compared with blank-virus group and blank control group, the concentrations of PGE2, Aggrecanase-1, and IL-1 significantly decreased and the concentration of IGF-1 significantly increased in the virus group (P<0.05), but the indicators in 3 groups were significantly higher than those in the normal control group (P<0.05). MRI showed that abnormal articular surface with high density could be found in virus group, blank-virus group, and blank control group, while the virus group had the minimum area. Gross observation and histological observation showed that the cartilage morphology of virus group, blank-virus group, and blank control group was accordance with early OA articular cartilage changes, but virus group was better than blank-virus group and blank control group in repair degree, whose improved Pineda score was significantly lower (P<0.05). Immunohistochemical staining showed that the virus group had deeper dyeing with occasional brown particles and more chondrocytes than blank-virus group and blank control group. By RT-PCR, COX-2 and Aggrecanase-1 mRNA expressions of cartilage in virus group were significantly decreased, and IGF-1 expression was significantly increased when compared with blank control group and the blank-virus group (P<0.05). Lentivirus-mediated multi-genes co-transfection in BMSCs can inhibit the expressions of COX-2 mRNA and Aggrecanase-1 mRNA, and enhance the IGF-1 mRNA expression, which decreases the concentration of inflammatory factors, and protects the joint cartilage effectively.

Effects of dietary postbiotic and inulin on growth performance, IGF1 and GHR mRNA expression, faecal microbiota and volatile fatty acids in broilers.

BackgroundPostbiotics (metabolic products by lactic acid bacteria) and prebiotics have been established as substitute to antibiotics in order to enhance immunity and growth performance in broiler chickens. Nonetheless, insufficient information is available on the effects of postbiotics and prebiotics combination on growth performance, faecal microbiota, pH and volatile fatty acids (VFA), as well as liver insulin like growth factor 1 (IGF1) and growth hormone receptor (GHR) mRNA expressions in broiler chickens. The aim of this experiment was to evaluate the effects of different types of postbiotics with different levels of prebiotic (inulin) on broiler for those parameters.ResultsThe results showed that birds fed T3: (0.3 % RI11 + 0.8 % Inulin), T4: (0.3 % RI11 + 1.0 % Inulin), and T6: (0.3 % RG14+ 1.0 % Inulin) had higher (p < 0.05) final body weight (BW) and total weight gain (WG) than other treatments. Birds fed T3 had lower feed conversion ratio (FCR) which was significantly different from those fed with negative control diet but was similar to other treatments. Postbiotic and inulin increased (p < 0.05) faecal lactic acid bacteria (LAB) and reduced (p < 0.05) Enterobacteriaceae count. Birds fed T4 and T6 had higher faecal acetic acid and propionic acid respectively, and both had higher total VFA and lactic acid bacteria but lower pH and Enterobacteriaceae (ENT) counts compared to other treatments. The liver of birds fed T4 and T6 had higher IGF1 expression compared to other treatments while T6 had higher GHR mRNA expression compared to other treatments.ConclusionsResults indicate that the addition of postbiotics and inulin combinations had beneficial effects on total BW, feed efficiency, mucosa architecture and IGF1 and GHR mRNA expression in broiler chickens.

Interaction between muscle and bone, and improving the effects of electrical muscle stimulation on amyotrophy and bone loss in a denervation rat model via sciatic neurectomy.

The side-to-side difference in bone mineral content and soft tissue composition of extremities and their associations have been observed in patients with stroke and the results are inconsistent. The aim of the present study was to investigate the interaction between bone mineral content (BMC), lean mass (LM) and fat mass (FM) in the paretic extremities in patients following stroke and to determine the effectiveness of electrical muscle stimulation (EMS) following sciatic neurectomy (SN) in rats. BMC, LM and FM were measured by dual-energy X-ray absorptiometry in 61 hemiplegic patients following stroke. In the rat model study, groups of 10 Sprague-Dawley rats were divided into EMS and non-EMS subgroups. Myostatin expression and tetracycline interlabel width were measured. There were significant decreases in BMC, LM and FM in paretic limbs compared to non-paretic limbs. Compared to non-EMS, downregulated myostatin mRNA, and upregulated mechano growth factor (MGF) and insulin-like growth factor 1 (IGF-1) mRNA expression levels were observed in the EMS subgroup (P<0.05). In conclusion, muscle may have an important role in maintaining BMC. EMS-induced muscle contraction effectively downregulated myostatin mRNA, upregulated MGF and IGF-1 mRNA expression in muscle fiber, and mitigated amyotrophy and cortical bone loss from SN.

Study on IGF-1 and IGFBP-3 gene expression in liver of young rats with intermittent hypoxia

Objective:Using the model of young rats with intermittent hypoxia, to study the expression of insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein-3 (IGFBP-3) in liver, and to provide objective evidence for the possible existence of growth retardation in children with OSAHS. Method:Divided 24 rats randomly into three groups, as control group and the mild and severe hypoxia group. Normally fed control group, without treatment, the other two groups were kept in the cabin has been simulated mild and severe intermittent hypoxia conditions of OSAHS in children. 8-hour day cycle of intermittent hypoxia, a total of 35 days. To the last day, animals were sacrificed and fresh liver tissues were fixed in liquid nitrogen, stored in the -80°C refrigerator, then took the molecular biology experiment of IGF-1 and IGFBP-3 in the liver. Result:The expression of IGF-1 mRNA in liver, semi-quantitative analysis showed that the difference of mRNA expression of IGF-1 among the three groups was statistically significant (P<0.05), pairwise comparison, mild hypoxia group was higher than control and severe group, there were significant differences (P<0.05), the control group and severe hypoxia had no significant difference (P>0.05). The expression of IGFBP-3 mRNA in liver showed that, differences between the three groups was not significant (P>0.05). IGF-1 expression in liver tissue analysis showed that, differences between the three groups was statistically significant (P<0.05), pairwise comparisons, its expression in the control group was lower than mild hypoxia group and severe group, there were significant differences (P<0.05), mild hypoxia and severe hypoxia group had no significant difference (P>0.05). IGFBP-3 expression in liver tissue analysis showed that, differences between the three groups was statistically significant (P<0.05), pairwise comparisons, the control group was higher than mild and severe hypoxic group, there were significant differences (P<0.05), mild hypoxia and severe hypoxia group had no significant difference (P>0.05). Conclusion:IGF-1 and IGFBP-3 in the liver of mild hypoxia group, severe hypoxic group and control group showed that their transcription levels and the severity of hypoxia had no significant correlation.

The Beneficial Impact of Antidepressant Drugs on Prenatal Stress-Evoked Malfunction of the Insulin-Like Growth Factor-1 (IGF-1) Protein Family in the Olfactory Bulbs of Adult Rats.

Insulin-like growth factor-1 (IGF-1) promotes the growth, differentiation, and survival of both neurons and glial cells, and it is believed to exert antidepressant-like activity. Thus, disturbances in the IGF-1 system could be responsible for the course of depression. To date, there have been no papers showing the impact of chronic antidepressant treatment on the IGF-1 network in the olfactory bulb (OB) in an animal model of depression. Prenatal stress was used as model of depression. Twenty-four 3-month-old male offspring of control and stressed mothers were subjected to behavioral testing (forced swim test). The mRNA expression of IGF-1 and IGF-1 receptor (IGF-1R) and the protein level of IGF-1 and its phosphorylation, as well as the concentrations of IGF-binding proteins (IGFBP-2, -4, -3, and -6), were measured in OBs before and after chronic imipramine, fluoxetine, or tianeptine administration. Adult rats exposed prenatally to stressful stimuli displayed not only depression-like behavior but also decreased IGF-1 expression, dysregulation in the IGFBP network, and diminished mRNA expression, as well as IGF-1R phosphorylation, in the OB. The administration of antidepressants normalized most of the changes in the IGF-1 system of the OB evoked by prenatal stress. These results suggested a beneficial effect of chronic antidepressant drug treatment in the alleviation of IGF-1 family malfunction in OBs in an animal model of depression.

Impact of periconceptional and preimplantation undernutrition on factors regulating myogenesis and protein synthesis in muscle of singleton and twin fetal sheep.

In this study, we determined the effect of maternal undernutrition in the periconceptional (PCUN: ∼80 days before to 6 days after conception) and preimplantation (PIUN: 0–6 days after conception) periods on the mRNA and protein abundance of key factors regulating myogenesis and protein synthesis, and on the relationship between the abundance of these factors and specific microRNA expression in the quadriceps muscle of singleton and twin fetal sheep at 135–138 days of gestation. PCUN and PIUN resulted in a decrease in the protein abundance of MYF5, a factor which determines the myogenic lineage, in singletons and twins. Interestingly, there was a concomitant increase in insulin-like growth factor-1 mRNA expression, a decrease in the protein abundance of the myogenic inhibitor, myostatin (MSTN), and an increase in the mRNA and protein abundance of the MSTN inhibitor, follistatin (FST), in the PCUN and PIUN groups in both singletons and twins. These promyogenic changes may compensate for the decrease in MYF5 protein abundance evoked by early embryonic undernutrition. PCUN and PIUN also increased the protein abundance of phosphorylated eukaryotic translation initiation factor binding protein 1 (EIF4EBP1; T70 and S65) in fetal muscle in singletons and twins. There was a significant inverse relationship between the expression of miR-30a-5p, miR-30d-5p, miR-27b-3p, miR106b-5p, and miR-376b and the protein abundance of mechanistic target of rapamycin (MTOR), FST, or MYF5 in singletons or twins. In particular, the expression of miR-30a-5p was increased and MYF5 protein abundance was decreased, in PCUN and PIUN twins supporting the conclusion that the impact of PCUN and PIUN is predominantly on the embryo.

애엽을 포함하는 해애탕의 에탄올 추출물이 제모된 C57BL/6 마우스의 발모에 미치는 영향

Objectives : The experiment was performed to investigate promotive effects of haeae-tang (HET) extract, a traditional Korean medicinal recipe, on hair growth, protein and gene expression in hair-removed C57BL/6. Methods : In experiment, animals were divided into 3 groups including normal (vehicle), HET ethanol extract and 5% minoxidil-treated groups (Minoxidil, positive control). The vehicle or testing samples were daily treated with 0.2ml per on hair-shaved dorsal skin of C57BL/6mice for 15 days. Effects of testing samples on hair growth was monitored through phototrichogram analysis by folliscope on the initial, <TEX>$5^{th}$</TEX>, <TEX>$10^{th}$</TEX>, <TEX>$15^{th}$</TEX> day, respectively. Also, gene and protein expressions of vascular endothelial growth factor (VEGF) and Insulin like growth factor-1 (IGF-1), relevant to hair growth, were examined. Results : Hair density and hair thickness of Minoxidil treated-group was significantly increased compared to that of vehicle application on the <TEX>$15^{th}$</TEX>day, respectively. Dorsal hair density of HET treated-group was significantly increased compared to that of vehicle application on the <TEX>$15^{th}$</TEX>day. In addition, the Minoxidil group significantly increased the expression of cutaneous IGF-1 protein and mRNA compared to that of the vehicle-applied group on the <TEX>$15^{th}$</TEX> day. And HET-treated group significantly increased the expression of dorsal VEGF protein compared to that of the vehicle-applied group on the <TEX>$15^{th}$</TEX> day. Conclusions : These results suggest that this Korean medicinal recipe, HET has promoting activity on hair growth in an Alopecia animal model thus it can be used as a material of agent or products for improvement or prevention of alopecia.

Administration of the inverse benzodiazepine agonist MRK-016 rescues acquisition and memory consolidation following peripheral administration of bacterial endotoxin

Recent evidence suggests that inflammation-induced decrements in cognitive function can be mitigated via manipulation of excitatory or inhibitory transmission. We tested the ability of the inverse benzodiazepine agonist, MRK-016 (MRK) to protect against LPS-induced deficits in memory acquisition and consolidation, using a contextual fear conditioning (CFC) paradigm. In Experiment One, mice received lipopolysaccharide (LPS) and/or MRK injections prior to CFC training, and were then tested 24h after training. In Experiment Two, animals received similar treatment injections immediately after training, and were tested 24h later. Additionally, hippocampal samples were collected 4h after LPS injections and immediately after testing, to evaluate brain-derived neurotrophic factor (BDNF) and insulin-like growth factor 1 (IGF-1) mRNA expression. Results indicate that MRK can protect against LPS-induced learning/memory decrements in both paradigms. We also found, in both paradigms, that animals treated with LPS/Saline expressed significantly less BDNF mRNA when compared to Saline/Saline-treated animals 4h after LPS administration, but that MRK did not restore BDNF expression levels. Further, treatment administrations had no effect on IGF-1 mRNA expression at any collection time-point. In summary, MRK-016 can protect against LPS-induced deficits in memory acquisition and consolidation, in this hippocampus-dependent paradigm, though this protection occurs independently of recovery of BDNF expression.

Correlation of insulin-like growth factor 1 expression in placenta with DNA methylation and fetal macrosomia

To explore the correlation between methylation of insulin-like growth factor 1 (IGF-1) gene promoter and its placenta-specific expression and fetal macrosoma. One hundred twenty nine healthy pregnant women were recruited between April 2011 and March 2012. Baseline data were collected with self-report questionnaires. Real-time quantitative PCR was used to determine the expression of IGF-1 mRNA in the placenta. Methylation level of the IGF 1 gene was determined with matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The expression of IGF-1 in placenta and its methylation level showed no significant difference between macrosomic fetuses and controls. No linear correlation was found between IGF-1 mRNA expression and methylation level of IGF-1 promoter (r=0.128, P=0.295). IGF-1 promoter region in placenta showed a hypomethylation status. However, a positive correlation was found between IGF-1 expression and birth weight below 4260 g (r=0.264, P=0.022). The expression of IGF-1 mRNA was significantly higher in those with a birth weight below 4260 g, which suggested that placental IGF-1 expression may contribute to increased birth weight. In regard to fetal overgrowth, however, there seemed to be a negative correlation in which placental IGF-1 expression was downregulated to limit fetal overgrowth. No linear correlation was found between placental IGF-1 expression and methylation level of IGF-1 promoter with a hypomethylation status. The contribution of placental IGF-1 expression to birth weight is bidirectional. Increased expression seems to promote fetal growth, while decreased expressions may curb overgrowth, therefore control fetal growth in a relatively normal range.

RNAi-mediated gene silencing of ST6GalNAc I suppresses the metastatic potential in gastric cancer cells.

ST6GalNAc I is a sialyltransferase controlling the expression of sialyl-Tn antigen (STn), which is overexpressed in several epithelial cancers, including gastric cancer, and is highly correlated with cancer metastasis. However, the functional contribution of ST6GalNAc I to development or progression of gastric cancer remains unclear. In this study, we investigated the effects of suppression of ST6GalNAc I on gastric cancer in vitro and in vivo. Gastric cancer cell lines were transfected with ST6GalNAc I siRNA and were examined by cell proliferation, migration, and invasion assays. We also evaluated the effect of ST6GalNAc I siRNA treatment in a peritoneal dissemination mouse model. The differences in mRNA levels of selected signaling molecules were analyzed by polymerase chain reaction (PCR) arrays associated with tumor metastasis in MKN45 cells. The signal transducer and activator of transcription 5b (STAT5b) signaling pathways that reportedly regulate the insulin-like growth factor-1 (IGF-1) were analyzed by Western blot. ST6GalNAc I siRNA inhibited gastric cancer cell growth, migration, and invasion in vitro. Furthermore, intraperitoneal administration of ST6GalNAc I siRNA- liposome significantly inhibited peritoneal dissemination and prolonged the survival of xenograft model mice with peritoneal dissemination of gastric cancer. PCR array confirmed that suppression of ST6GalNAc I caused a significant reduction in expression of IGF-1 mRNA. Decreased IGF-1 expression in MKN45 cells treated with ST6GalNAc I siRNA was accompanied by reduced phosphorylation of STAT5b. ST6GalNAc I may regulate the gene expression of IGF-1 through STAT5b activation in gastric cancer cells and may be a potential target for treatment of metastasizing gastric cancer.

Regulatory effects of intermittent noxious stimulation on spinal cord injury-sensitive microRNAs and their presumptive targets following spinal cord contusion.

Uncontrollable nociceptive stimulation adversely affects recovery in spinally contused rats. Spinal cord injury (SCI) results in altered microRNA (miRNA) expression both at, and distal to the lesion site. We hypothesized that uncontrollable nociception further influences SCI-sensitive miRNAs and associated gene targets, potentially explaining the progression of maladaptive plasticity. Our data validated previously described sensitivity of miRNAs to SCI alone. Moreover, following SCI, intermittent noxious stimulation decreased expression of miR124 in dorsal spinal cord 24 h after stimulation and increased expression of miR129-2 in dorsal, and miR1 in ventral spinal cord at 7 days. We also found that brain-derived neurotrophic factor (BDNF) mRNA expression was significantly down-regulated 1 day after SCI alone, and significantly more so, after SCI followed by tailshock. Insulin-like growth factor-1 (IGF-1) mRNA expression was significantly increased at both 1 and 7 days post-SCI, and significantly more so, 7 days post-SCI with shock. MiR1 expression was positively and significantly correlated with IGF-1, but not BDNF mRNA expression. Further, stepwise linear regression analysis indicated that a significant proportion of the changes in BDNF and IGF-1 mRNA expression were explained by variance in two groups of miRNAs, implying co-regulation. Collectively, these data show that uncontrollable nociception which activates sensorimotor circuits distal to the injury site, influences SCI-miRNAs and target mRNAs within the lesion site. SCI-sensitive miRNAs may well mediate adverse consequences of uncontrolled sensorimotor activation on functional recovery. However, their sensitivity to distal sensory input also implicates these miRNAs as candidate targets for the management of SCI and neuropathic pain.

Rosiglitazone suppresses angiogenesis in multiple myeloma via downregulation of hypoxia-inducible factor-1α and insulin-like growth factor-1 mRNA expression.

Rosiglitazone (RGZ) is a thiazolidinedione ligand of peroxisome proliferator-activated receptor-γ. Our previous studies have confirmed that RGZ possesses antitumoral properties. Bone marrow angiogenesis exhibits an important role in multiple myeloma (MM), and angiogenesis often correlates with the prognosis and disease burden of MM. However, to the best of our knowledge, inhibition of angiopoiesis by RGZ in MM has not yet been reported. The present study aimed to investigate whether RGZ prevents angiogenesis and the possible underlying mechanism of this effect in MM. RPMI‑8226 cells, primary myeloma cells from patients with MM or mononuclear cells from healthy patients were treated with different concentrations of RGZ, and various biological responses were detected using MTT, reverse transcription-polymerase chain reaction and western blot assays. The expression levels of hypoxia-inducible transcription factor‑1α (HIF1α) and insulin-like growth factor‑1 (IGF1) were significantly increased in the RPMI-8226 cells and the primary myeloma cells from the patients with MM compared with those in the mononuclear cells from the healthy patients. The results also showed that RGZ was able to inhibit proliferation and reduce viability of RPMI-8226 cells in a concentration-and time-dependent manner. RGZ was able to concentration‑dependently inhibit the expression of HIF1α and IGF1 mRNA in RPMI-8226 and primary myeloma cells from patients with MM. RGZ also inhibited the expression of pAKT and downregulated the expression levels of phosphorylated extracellular signal-regulated kinase (ERK) in RPMI-8226 cells. The results suggested that RGZ inhibits the angiopoiesis of tumors by interfering with the phosphatidylinositol 3-kinase/AKT and ERK signaling pathways.