IGF-I Expression Research Articles

Muscle regeneration in gilthead sea bream: Implications of endocrine and local regulatory factors and the crosstalk with bone.

Fish muscle regeneration is still a poorly known process. In the present study, an injury was done into the left anterior epaxial skeletal muscle of seventy 15g gilthead sea bream (Sparus aurata) juveniles to evaluate at days 0, 1, 2, 4, 8, 16 and 30 post-wound, the expression of several muscle genes. Moreover, transcripts' expression in the bone (uninjured tissue) was also analyzed. Histology of the muscle showed the presence of dead tissue the first day after injury and how the damaged fibers were removed and replaced by new muscle fibers by day 16 that kept growing up to day 30. Gene expression results showed in muscle an early upregulation of igf-2 and a downregulation of ghr-1 and igf-1. Proteolytic systems expression increased with capn2 and ctsl peaking at 1 and 2 days post-injury, respectively and mafbx at day 8.A pattern of expression that fitted well with active myogenesis progression 16 days after the injury was then observed, with the recovery of igf-1, pax7, cmet, and cav1 expression; and later on, that of cav3 as well. Furthermore, the first days post-injury, the cytokines il-6 and il-15 were also upregulated confirming the tissue inflammation, while tnfα was only upregulated at days 16 and 30 to induce satellite cells recruitment; overall suggesting a possible role for these molecules as myokines. The results of the bone transcripts showed an upregulation first, of bmp2 and ctsk at days 1 and 2, respectively; then, ogn1 and ocn peaked at day 4 in parallel to mstn2 downregulation, and runx2 and ogn2 increased after 8 days of muscle injury, suggesting a possible tissue crosstalk during the regenerative process. Overall, the present model allows studying the sequential involvement of different regulatory molecules during muscle regeneration, as well as the potential relationship between muscle and other tissues such as bone to control musculoskeletal development and growth, pointing out an interesting new line of research in this group of vertebrates.

The effects of combined or singular administration of formic acid and Pediococcus acidilactici on stress resistance, growth performance, immune responses and related genes expression in common carp, Cyprinus carpio

In the current study, the effects of dietary formic acid and probiotic supplementation in growth, serum immune parameters, salinity tolerance, and gene expression of common carp, Cyprinus carpio were studied. 360 healthy common carp with average weight of (5.54 ± 1.08 g) were randomly stocked into 12 fiberglass tanks with 300 L capacity having 30 individuals in each tank. The fish were fed for 56 days with four experimental diets i.e., Control (Formic acid 0 + Pediococcus acidilactici 0), FA (Formic acid 0.3% + Probiotic 0), Pro (Formic acid 0 + P. acidilactici 0.1%), and FA+Pro (Formic acid 0.3% + P. acidilactici 0.1%). Growth performance, serum immune parameters, and gene expression of the experimental fish were examined at the completion of the feeding study. Fish fed the combination of formic acid and probiotic (FA+Pro) caused significantly (P < 0.05) higher weight gain and lower feed conversion ratio compared to the control and the experimental diets. Specific growth rate didn’t show significant differences among experimental diets (P > 0.05). Fish administered formic acid and probiotics (FA+Pro) also had significantly higher serum immune parameters (total protein, globulin, and lysozyme) than the control (P < 0.05). Moreover, the relative expression of GH, IGF1, Lyz, and IL-1 genes increased significantly (P < 0.05) with combined supplementation when compared to fish fed just formic acid or probiotics and the control. During 7 days of exposure to saline conditions, fish fed with a combination of formic acid and/or probiotic showed a significantly higher survival rate (P < 0.05). It can be concluded that combined administration of P. acidilactici as probiotic with an organic acid (FA) exert much more beneficial effects rather than singular administration. Therefore, it can be considered as promising feed additive for carp culture.

Inhibiting miR-186-5p relieves traumatic brain injury by regulating insulin-like growth factor-I-NLRP3/ASC/caspase-1 signaling pathway.

Previous studies have shown that micro-RNA (miR)-186-5p can affect apoptosis of cells by regulating insulin-like growth factor-I (IGF-1). However, the role of miR-186-5p-IGF1 axis in traumatic brain injury (TBI), especially oxidative stress and neuroinflammatory response, remains to be further studied. Lipopolysaccharide (5 μg/mL) was used to activate microglia in vitro. The expression of miR-186-5p, IGF-1 was detected by quantitative reverse transcription PCR (qRT-PCR). ELISA and western blot were used to detect the inflammatory factors and oxidative stress. Western blot was used to detect apoptotic proteins (Bax, Bcl2 and C-caspase3), inflammatory proteins (iNOS and COX2), oxidative stress proteins (Nrf2 and HO-1) and NLRP3/apoptosis-associated speck-like protein containing a CARD (ASC)/caspase-1 inflammatory bodies. MiR-186-5p inhibitor could reduce the inflammatory factors and oxidative stress in BV2 treated with lipopolysaccharide, and reduce apoptosis. In addition, we also found that inhibition of miR-186-5p increased the expression of IGF-1, which is necessary for nervous system development. Luciferase activity assay confirmed that IGF-1 was the direct target gene of miR-186-5p. Inhibiting miR-186-5p, through upregulation IGF-1, attenuates the inflammatory factors, oxidative stress and by inhibiting NLRP3/ASC/caspase-1 signal pathway TBI in-vitro model.

Resveratrol regulates insulin resistance to improve the glycolytic pathway by activating SIRT2 in PCOS granulosa cells.

Insulin resistance (IR) has a close relationship with the main clinical manifestations of patients with PCOS; hence, the research and development of new drugs to treat PCOS by improving IR is a desiderate task at present. Resveratrol (RES) possesses a variety of beneficial pharmacological functions, such as antioxidation, anti-inflammatory, regulating glucose, and lipid metabolism. However, whether RES could improve IR and the underlying mechanisms remained unclear in PCOS. SD rats received a high-fat diet and letrozole for 30 days to establish the PCOS model and then intervened with RES for 30 days. The results demonstrated that RES played a protective role on the IR in PCOS rats, which significantly decreased the levels of blood glucose and serum insulin, up regulated the expression of IGF1R, and down regulated the expression of IGF1. In vitro, KGN cells were treated with insulin, RES, and AGK2, respectively. We found that a high dose of insulin (4μg/mL) significantly inhibited KGN cell viability, decreased the level of lactic acid, and increased the level of pyruvate, while RES (25μM) attenuated the growth-inhibitory effect, as well as increased the level of lactic acid and decreased the level of pyruvate after high levels of insulin treatment. Simultaneously, RES up regulated the expression level of the crucial rate-limiting enzymes relating to glycolytic pathways, such as LDHA, HK2, and PKM2. Furthermore, AGK2 remarkably inhibited the expression level of SIRT2, which was similar to the same negative effects processed by insulin. Meanwhile, RES overtly repaired the glycolysis process by reversing the levels of lactic acid and pyruvate, together with up regulating the expression level of LDHA, HK2, and PKM2, after AGK2 treatment. RES could effectively improve insulin resistance and restore the glycolysis pathway by regulating SIRT2, which may contribute to attenuating the ovarian damage of PCOS rats and provide a potential treatment for patients with PCOS.

Mechanism of autophagy mediated by IGF-1 signaling pathway in the neurotoxicity of lead in pubertal rats

Lead can damage neuron synapses in the hippocampus and cause synaptic plasticity losses, and learning, memory, and intelligence impairments. Previous studies have focused on the functional and structural plasticity of hippocampal synapses; however, the specific molecular mechanisms behind such impairments are not fully understood. This study aimed to elucidate the molecular mechanisms of cognitive impairment in rats following chronic lead exposure and mitigate or prevent lead toxicity in the central nervous system. We found that lead exposure caused significant damage to rat nervous systems, that is, compared with the control group, the lead treatment group had more autophagosomes in their hippocampal neurons; lower serum and hippocampal IGF-1 levels; lower hippocampal IGF-1, IGF-1R, PI3K, Akt, and mTOR gene expression; and upregulated hippocampal autophagy-associated proteins levels. Brain stereotactic technology was used to conduct autophagy inhibitor in vivo intervention experiments, and the results of these experiments suggest that the autophagy inhibitor DC661 inhibited lead-exposure-induced autophagy and autophagy-related gene expression in the rat hippocampus, possibly through activation of the IGF-1 pathway. Overall, our findings suggest that lead might activate hippocampal autophagy through the IGF-1/PI3K/Akt/mTOR signaling pathway. Therefore, this study provides a novel molecular mechanism underlying developmental toxicity in pubertal rats induced by lead exposure and provides a new target for anticipation and reversal of such neurotoxicity.

Ewe early gestation supplementation with eicosapentaenoic and docosahexaenoic acids affects the liver, muscle, and adipose tissue fatty acid profile and liver mRNA expression in the offspring.

Our objectives were to assess the effects of eicosapentaenoic (EPA) and docosahexaenoic acids (DHA) supplementation to pregnant ewes during the first third of gestation on their offspring's liver, adipose, and muscle tissues fatty acid (FA) profile and liver mRNA expression after a finishing period receiving diets with different FA profiles. Twenty-four post-weaning lambs, blocked by sex and body weight, were used in a 2 × 2 factorial arrangement of treatments. The first factor was dam supplementation (DS) in the first third of gestation with 1.61% of Ca salts of palm fatty acid distillate (PFAD) or Ca salts enriched with EPA-DHA. Ewes were exposed to rams with marking paint harnesses during the breeding. Ewes started DS at the day of mating, considered day 1 of conception. Twenty-eight days after mating, ultrasonography was used to confirm pregnancy, and nonpregnant ewes were removed from the groups. After weaning, the offspring lambs were supplemented (LS, second main factor) with two different FA sources (1.48% of PFAD or 1.48% of EPA-DHA) during the growing and fattening phase. Lambs were fed the LS diet for 56 d and sent to slaughter, where the liver, muscle, and adipose tissue samples were collected for FA analysis. Liver samples were collected for relative mRNA expression for genes associated with FA transport and metabolism. The data were analyzed as a mixed model in SAS (9.4). In the liver, the amount of C20:5 and C22:6 (P < 0.01) increased in lambs with LS-EPA-DHA, while some C18:1 cis FA isomers were greater in the lambs from DS-PFAD. In muscle, amounts of C22:1, C20:5, and C22:5 increased (P < 0.05) in lambs born from DS-EPA-DHA. The adipose tissue amounts of C20:5, C22:5, and C22:6 were greater (P < 0.01) in lambs from LS-EPA-DHA. Interactions (DS × LS; P < 0.05) were observed for DNMT3β, FABP-1, FABP-5, SCD, and SREBP-1; having greater mRNA expression in liver tissue of LS-EPA-DHA, DS-PFAD and LS-PFAD, DS-EPA-DHA lambs compared with the lambs in the other two treatments. Liver ELOVL2 mRNA relative expression (P < 0.03) was greater in the offspring of DS-PFAD. Relative mRNA expression (P < 0.05) of GLUT1, IGF-1, LPL, and PPARγ increased in the liver from LS-EPA-DHA lambs. Dam supplementation during early gestation using with different FA sources changed the lipid FA profile in MT, LT, and SAT during the finishing period depending on the tissue and type of FA source administered during the growing phase.

The effect of aerobic training and curcumin supplementation on the expression of IGF-1 gene in muscle rat

Background: The purpose of this research was to investigate the effect of four weeks of moderate aerobic training and nanocurcumin supplementation on IGF-1 gene expression and its receptor in the soleus muscle of rats. Materials and Methods: 32 male Wistar rats, approximately eight weeks old, with a weight range of 200±10 grams, were prepared and kept for one week under standard conditions of light, temperature, and humidity. Rats were randomly divided into 4 groups of 8 series: healthy control group, aerobic training group, nanocurcumin group and aerobic training + nanocurcumin group. The training protocol was for 4 weeks, 5 days a week, in the first week, 20 minutes of activity with a speed of 18 meters per minute, and finally in the 4th week, it reached 35 minutes of activity with a speed of 20 meters per minute. Nanocurcumin supplement was also used for each animal at 80 mg / kilogram of body weight. One-way analysis of variance and Bonferroni's post hoc test was used at the significance level (P<0.05). Results: The results showed an increase in IGF-1 and IGF-1R gene expression levels in the soleus muscle in all groups compared to the control group. Conclusion: The combination of training and nanocurcumin supplementation probably increases the antioxidant capacity of the body and causes a greater increase in the expression of the IGF-1 gene and IGF-1R in skeletal muscles, which indicates more anabolic effects.

The Endocrine Approach of Melanoma: The Puzzle of Estrogen Receptors Expression

"Melanoma outcome seems different between females and males, with a potential protective role of estrogen (E) through estrogen receptors (ER) expression into the tumor. In the study of ERs, both alfa (ERα) and beta (ERβ) is a well-known endocrine elements in non-melanoma tumors, like mammary and endometrial cancer. Immunohistochemistry (IHC) assessment of melanoma concerning ERs represents a path to explore the tumor profile to provide useful information concerning the prognostic and potential adjuvant treatment. Currently, this is not a routine practice, nor a mandatory step for deciding the medical therapy. Typically, IHCs are based on usual kits for mammary tumors regarding ERs configuration. Prior/concomitant use of oral contraceptives and hormonal replacement therapy is not correlated with a better prognostic in melanoma; neither have they represented a contraindication for survivors of melanoma; a subset of tumors might present a higher ER expression which is potentially targeted by the hormone-based treatment as SERMs (Selective Estrogen Receptors Modulator), for instance, tamoxifen. Experimental studies on melanoma cell lines confirmed the anti-tumor activity of ERβ which might function as a prognostic marker. G-protein-coupled estrogen receptors in melanocytes and keratinocytes might be involved, too. Additional crosstalk of TGF-β (Transforming Growth Factor β), respective IGF1 (Insulin-like Growth Factor), and ERα expression are involved in tumorigenic pathways. Recent preclinical studies showed the potential benefits of diarylpropionitrile, a selective agonist of ERβ; pyrazole derivates 21-23 can block ERs. Murine melanoma models showed the interference of anti-estrogenic medication (like molecule fulvestrant) to enhance immune checkpoint blockade, a modern approach to solid cancers. The proliferation of melanoma might be partially explained by ERs; whether this is generally applicable or there is a subgroup of tumors particularly related to E status is still debatable. The subject of E status in melanoma is far from clear at this point and further studies are necessary concerning this particular issue to implement it as a practical approach in the daily management of a disease that still has a very severe prognostic nowadays "

Impact of Crossbreeding Between Local and Foreign Strains on the Growth Traits and the Expression of Insulin-Like Growth Factor Genes on Their Chicks

Abstract: The aim of this work was to evaluate the impact of crossing pure Gimmizah or Bandara (local breeds) with pure Ross (a foreign breed) on growth rate, and the expression of IGF genes. Two local breeds, Gimmizah (G) and Bandara (B), plus one exotic strain of meat-type Ross 308 (R) parents, were used. Combinations of these genotypes were tested on a total of 120 birds from three pure parents. The 180 hatched chicks were divided into three purebreds G×G, B×B, and R×R and three crossbreds R×G, R×B, and G×R and reared for 7 weeks. Body weight and weight gain were recorded for each genetic group. The feed intake and feed conversion ratio were calculated. At the 7th week of age, tissue samples from breast muscle and pituitary gland were collected for IGF gene mRNA expression analysis. The results revealed that the R×R chicks displayed significantly higher final body weight, average daily weight gain, and average daily feed intake when compared to the G×G, B×B, and other crossbred chick lines. While both R×B and G×R crossbred chicks had significantly higher final body weights and average daily weight gains than R×G crossbred chicks and local chicks (G×G and B×B). A better feed conversion ratio was observed in G×R, B×B, R×R and R×B chicks. Crossbred lines (R×B and G×R) had higher dressing percentages than R×G and local breeds. The highest muscular expression of the IGF-II gene was recorded in R×G chicks, while the other crossbred lines, R×B and G×R expressed higher levels of the IGF-II gene than purebred lines. In relation to IGF-I*, R×B chicks had higher relative expression of this gene than other crossbred and purebred lines. When compared to other crossbred lines (R×B and G×R), the expression of IGF-I in brain tissues was significantly higher in the R×G crossbred line. IGF-II and IGF-I* expression levels in brain tissue were higher in the R×G chicks compared to the G×R, G×G, B×B, and R×R chicks. It was concluded that crossing pure Gimmizah or Bandara, as local breeds, with pure Ross 308, as a foreign breed, improved growth traits in the obtained crossbred chicks.

Revisiting Skeletal Muscle Dysfunction and Exercise in Chronic Obstructive Pulmonary Disease: Emerging Significance of Myokines.

Skeletal muscle dysfunction (SMD) is the most significant extrapulmonary complication and an independent prognostic indicator in patients with chronic obstructive pulmonary disease (COPD). Myokines, such as interleukin (IL)-6, IL-15, myostatin, irisin, and insulin-like growth factor (IGF)-1, play important roles in skeletal muscle mitochondrial function, protein synthesis and breakdown balance, and regeneration of skeletal muscles in COPD. As the main component of pulmonary rehabilitation, exercise can improve muscle strength, muscle endurance, and exercise capacity in patients with COPD, as well as improve the prognosis of SMD and COPD by regulating the expression levels of myokines. The mechanisms by which exercise regulates myokine levels are related to microRNAs. IGF-1 expression is upregulated by decreasing the expression of miR-1 or miR-29b. Myostatin downregulation and irisin upregulation are associated with increased miR-27a expression and decreased miR-696 expression, respectively. These findings suggest that myokines are potential targets for the prevention and treatment of SMD in COPD. A comprehensive analysis of the role and regulatory mechanisms of myokines can facilitate the development of new exercise-based therapeutic approaches for patients with COPD.

Nine kinds of novel cytokines identified in children with lead exposure

Lead (Pb) is a neurotoxic heavy metal element with many recognized adverse health side effects, and its main target of lead toxicity is the central nervous system. The mechanism of lead toxicity is still uncertain. However, there are few studies investigated the cytokines changes caused by lead exposure in children. The BLLs was quantified using flame atomic absorption spectroscopy. The novel cytokines were detected by RayBio@ Human Cytokine Antibody Array. A total of 4 children with elevated blood lead levels (BLLs) and 4 children with low BLLs were chosen in the study. Volcano plot analysis was performed to identify significant proteins, with the criteria: P value &lt;0.05 and log2 fold change &gt;1. The mean BLLs of children with elevated BLLs (5.675±1.018 μg/dL) has significant difference compared with those with low BLLs (1.975±0.3966 μg/dL) (P=0.0148, t=3.385). And 9 kinds of novel cytokines were identified. The expression of IL-6, IL-8 and IL-17 was significantly up-regulated, while the expression of BDNF, BMP-4, IGF-1, IL-7, IL-10 and Leptin was significantly down-regulated.

Oral Administration of Spirulina platensis at Early Gestation Modulates Litter Size and the Expression of Inhibin, Insulin, IGF-I, CO Q10, and BMP-15 in Ewes Induced for Twinning.

Gestation in sheep necessitates the support of nutrients to avoid early embryonic mortalities. Therefore, this study investigates the effects of supplementing either L-arginine or Spirulina alga in the first trimester on the pregnancy rate, litter size, inhibin, insulin, IGF-I, CO Q10, and bone morphogenetic peptide 15 (BMP15) in maternal circulation. Animals were offered barley (500 g/head/day, 14% CP), alfalfa hay (1 kg/head/day, 12% CP), clean water, and balanced salt block licks as free choices. Forty Noemi and Najdi ewes were randomly allotted into three groups: control (C, n = 8), L-arginine (ARG, n = 16), and Spirulina (SP, n = 16). All females were implanted with CIDR for ten days. On days 8, 9, and 10, treated ewes were given a protocol comprised of human recombinant FSH at descending doses (50, 50, 40, 40, and 30, 30 IU, A.M, and P.M, respectively). At the fifth dose, animals were given an equivalent dose of hCG (240 IU). After CIDR withdrawal, ewes were exposed to fertile rams for mating. SP-ewes were orally given 50 ml (2%) Spirulina, and ARG-ewes were given 50 ml (35 mg/kg BW) L-arginine daily for 50 days postbreeding. Blood inhibin, insulin, IGF-I, CO Q10, and BMP15 were determined throughout gestation until parturition. The findings indicated that the conception rates were 25, 75, and 87.5% in C, ARG, and SP, respectively (P < 0.05). The percent of ewes giving birth to twins was 0, 25, and 50% in C, ARG, and SP, respectively (P < 0.05). The survival rates were 100, 81.8, and 83.3%, respectively. Birth weight was 5.6, 3.2, and 3.4 kg in C, ARG, and SP, respectively. Weaning weights were 28.3, 25.6, and 27.2 kg in C, ARG, and SP, respectively. BMP-15 was reduced (P < 0.05) in ARG than in C and SP. However, SP decreased (P < 0.05) inhibin more than in C and ARG. ARG and SP increased (P < 0.05) insulin than in C, whereas SP decreased (P < 0.05) IGF-I. SP increased CO Q10 compared with ARG. Ewes bearing twins revealed higher (P < 0.05) IGF-I (8.57 ng/ml) than those bearing singles (4.63 ng/ml); however, BMP-15 was higher in single (796.6 pg/ml) than in twin-bearing (387.5 pg/ml) ewes. In conclusion, providing early-gestating ewes with Spirulina enhances maternal health, productivity, and reproductive outcomes.

Abstract C064: IGF-II regulates a critical mitochondrial protein, Magmas, with differential expression between African American and Non-Hispanic White women with TNBC

Abstract Breast cancer is the most common malignancy in women. Young African American (AA) women are affected by a higher incidence and mortality rate of triple-negative breast cancer (TNBC) compared to Non-Hispanic White (NHW) women. Currently, available treatment options are suboptimal, with a 5-year survival of less than 12% for metastatic TNBC. Our laboratory demonstrated that IGF-2 plays a pivotal role in the tumor aggressiveness of AA women. Moreover, tissues from AA women express high levels of IGF-2 and exhibit high activation of IGF-2 signaling pathways associated with tumorgenesis and chemoresistance. An emerging pathway linked to IGF-2 mediated chemoresistance and cell survival in TNBC is the regulation of mitochondrial and survival proteins. Mitochondrial Magmas is a mitochondrial protein found in all eukaryotic cells that is required for cell viability. It functions to transport nuclear proteins from the inner mitochondrial membrane to the mitochondrial matrix. Magmas can buffer increased reactive oxidative species (ROS) and promote survival of cancer cells. Overexpression of Magmas reduced ROS and enhanced cellular tolerance to oxidative stress; however, its downregulation raised cellular ROS and made cells more vulnerable to ROS-mediated death. Given the role of IGF-2 in regulating mitochondrial proteins, we investigated if IGF-2 regulated Magmas. We hypothesize that IGF-2 increases Magmas, and since AA women express higher levels of IGF-2, they will express higher Magmas levels. We used the CRL-2335 AA TNBC cell line and an IGF-2 knockdown of those same cells. Western blotting was used to detect the specific isoforms of IGF-2 and Magmas from cell lysates. Immunohistochemistry (IHC) characterized Magmas expression in paired, normal-tumor breast cancer tissues. Real time PCR was used to evaluate the expression of IGF2 and MAGMAS. The preliminary results showed that IGF-2 knockdown in CRL-2335 AA TNBC cells significantly decreased Magmas protein expression and its corresponding mRNA. In addition, tissue analysis demonstrated high levels of Magmas staining in TNBC tissue while little or no staining was detected in the normal tissues. Furthermore, tumor tissues of AA women showed significantly higher expression of Magmas as compared to tissues from NHW women. Targeting Magmas represents a novel and much needed tool to treat TNBC. Our team is currently validating Magmas as a target for Theranostics treatment, a promising emerging radiotherapy for TNBC patients. Citation Format: Jennifer D. Tran, Alfonso D. Duran, Qian-Wei Tan, Frankis G. Almaguel, Daisy De León. IGF-II regulates a critical mitochondrial protein, Magmas, with differential expression between African American and Non-Hispanic White women with TNBC [abstract]. In: Proceedings of the 15th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2022 Sep 16-19; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr C064.

Characteristic amino acid residues in the growth hormone receptor gene on Mus minutoides underlying dwarfism.

The African pygmy mouse ( Mus minutoides ) displays a dwarfism phenotype distinctive from closely related species. This study aimed to investigate the growth hormone receptor (Ghr) gene sequence in M. minutoides . We identified several amino acid variations, including the P469L mutation. Our findings suggest that this mutation affects Ghr protein functionality, decreasing Igf1 expression and contributing to the dwarfism observed in M. minutoides . Further studies utilizing genome editing technology are necessary to elucidate the mechanisms involved in mammalian body size determination.

Concentraciones del factor de crecimiento similar a la insulina 1 (IGF-1) en el líquido sinovial de caballos sanos y osteoartríticos, y su correlación con las citoquinas proinflamatorias IL-6 y TNF

Insulin-like growth factor I (IGF-1) is the most important known growth factor for cartilage repair in horses. It promotes mitosis of chondrocytes, collagen II expression, and extra cellular matrix production. Osteoarthritis (OA) is the most common musculoskeletal condition that causes lameness and poor performance in sport horses. A total of 11 lame horses were clinically and radiographically evaluated, and all were confirmed to suffer a front metacarpophalangeal lameness by a positive flexion test, a low-4-point nerve block and an intraarticular block. Total protein, IGF-1, IL-6 and TNFa were determined by ELISA, demonstrating changes and different correlations between clinical condition, radiographic changes and degree of inflammation. All horses with joint associated pain and therefore associated lameness, demonstrated a significant increase of total protein (P&lt;0.0001) and IGF-1 concentration (P&lt;0.05). Concentrations of IL-6 and TNFa between controls and lame horses demonstrated significant differences (P&lt;0.01 and P&lt;0.001 respectively). Horses with less radiographic changes, demonstrated the highest IGF-1 expression in synovial fluid, and horses with more chronic OA conditions had very similar IGF-1 expression levels than control joints. In all lame joints, it was identified by Western blot a lighter isoform of IGF-1 (~7.5 kDa) which was inflammation related and it is the molecular weight of the mature peptide, and all control joints expressed a heavier isoform (~12 kDa). This finding could lead to new research for sequencing and targeting the isoform which is not expressed during an inflammatory process within a joint, and to have a better understanding of its role in the horse’s joint.

The effective constituent puerarin, from Pueraria lobata, inhibits the proliferation and inflammation of vascular smooth muscle in atherosclerosis through the miR-29b-3p/IGF1 pathway

Context Atherosclerosis (AS) is the main cause of cardiovascular and cerebrovascular diseases. Pueraria lobata (Willd.) Ohwi (Fabaceae) has a positive effect on improving these diseases. Objective The P. lobata effect on the proliferation and inflammation of vascular smooth muscle in AS and the potential mechanism were investigated. Materials and methods By feeding a high-fat diet to 8-week-old apolipoprotein E knockout mice, an atherosclerosis model was created. H&E and IHC staining were used to analyse the histopathology of mice. CCK-8, TUNEL, and scratch tests were used to detect cell proliferation, apoptosis, and migration after 24 h treatment, respectively. ELISA was performed to evaluate the level of IL-6 and IL-8. The target miRNA and its downstream target gene were screened by the bioinformatics method; RT-qPCR has conducted to analyse the expression of these genes. Results In the aortic tissue and serum of AS mice, puerarin can lower the expression of α-SMA and the inflammatory proteins IL-6 and IL-8. Puerarin (200 M) decreased hVSMC proliferation, migration, and IL-6 and IL-8 secretion by more than half. The inhibitory impact of puerarin on hVSMC was decreased by overexpression of miR-29b-3p. IGF1 was miR-29b-3p's downstream target gene. IGF1 expression increased almost 3-fold in AS mice and hVSMC, but miR-29b-3p mimic inhibited it. The effect of miR-29b-3p on hVSMC was reversed when IGF1 was overexpressed. Discussion and conclusions Puerarin inhibits the proliferation and inflammation of vascular smooth muscle in AS through the miR-29b-3p/IGF1 pathway. Puerarin may have a beneficial effect in the treatment of atherosclerosis and offer a novel therapy option.

Protective and sex-specific effects of moderate dose folic acid supplementation on the placenta following assisted reproduction in mice.

Epigenetic defects induced by assisted reproductive technologies (ART) have been suggested as a potential mechanism contributing to suboptimal placentation. Here, we hypothesize that ART perturbs DNA methylation (DNAme) and gene expression during early placenta development, leading to abnormal placental phenotypes observed at term. Since folic acid (FA) plays a crucial role in epigenetic regulation, we propose that FA supplementation can rescue ART-induced placental defects. Female mice were placed on a control diet (CD), a moderate 4-fold (FAS4) or high dose 10-fold (FAS10) FA-supplemented diet prior to ART and compared to a natural mating group. ART resulted in 41 and 28 differentially expressed genes (DEGs) in E10.5 female and male placentas, respectively. Many DEGs were implicated in early placenta development and associated with DNAme changes; a number clustered at known imprinting control regions (ICR). In females, FAS4 partially corrected alterations in gene expression while FAS10 showed evidence of male-biased adverse effects. DNAme and gene expression for five genes involved in early placentation (Phlda2, EphB2, Igf2, Peg3, L3mbtl1) were followed up in placentas from normal as well as delayed and abnormal embryos. Phlda2 and Igf2 expression levels were lowest after ART in placentas of female delayed embryos. Moreover, ART concomitantly reduced DNAme at the Kcnq1ot1 ICR which regulates Phlda2 expression; FAS4 partially improved DNAme in a sex-specific manner. In conclusion, ART-associated placental DNAme and transcriptome alterations observed at mid-gestation are sex-specific; they may help explain adverse placental phenotypes detected at term and are partially corrected by maternal moderate dose FA supplementation.

MiR-216b-5p regulates proliferation and apoptosis of ox-LDL-stimulated VSMCs and HUVECs via IGF2.

Atherosclerosis (AS) is one of the principal causes of cardiovascular disorder. Reportedly, vascular smooth muscle cells (VSMCs) and human umbilical vein endothelial cells (HUVECs) play key roles in AS development, and microRNAs (miRNAs) regulate their functions. The function of miR-216b-5p in AS remains unknown. Human VSMCs and human HUVECs were treated with ox-LDL to establish the in vitro model of AS. MiR-216b-5p and IGF2 expressions in VSMCs and HUVECs were probed by qRT-PCR and western blot. The viability, cell cycle progression, and apoptosis of VSMCs and HUVECs were evaluated by Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine, and flow cytometry assays, respectively. The binding sites between IGF2 3'UTR and miR-216b-5p were validated by dual-luciferase reporter assay. miR-216b-5p expression was declined in ox-LDL-induced VSMCs and HUVECs. In VSMCs, miR-216b-5p overexpression inhibited excessive proliferation and induced apoptosis. MiR-216b-5p could markedly restrain the viabiblity of VSMCs induced by ox-LDL and enhanced the viability of HUVECs. Additionally, IGF2 was confirmed as the direct target of miR-216b-5p and transfection of IGF2 overexpression plasmids rescued the effects of miR-216b-5p on VSMCs and HUVECs. miR-216b-5p alleviates the dysfunction of VSMCs and HUVECs caused by ox-LDL via repressing IGF2, and exerts protective functions to block the development of AS.

Interrelationships Between miR-34a and FSH in the Control of Porcine Ovarian Cell Functions.

Our study aimed to elucidate the effect of miR-34a mimics and miR-34a inhibitor and their combination on basic functions of ovarian cells cultured with and without FSH, and effect of FSH on expression of endogenous miR-34a. Viability, proliferation, proportion of proliferative active cells, apoptosis, proportion of DNA fragmented cells, accumulation of FSHR, steroid hormones, IGF-I, oxytocin, and prostaglandin E2 release, and expression levels of miR-34a were analysed. MiR-34a mimics decreased proliferation, apoptosis, testosterone, and estradiol output, stimulated release of progesterone, IGF-I, oxytocin, and occurrence of FSHR. MiR-34a inhibitor had an opposite effect and prevented effects of miR-34a mimics. FSH promoted expression of miR-34a, viability, proliferation, steroid hormones, IGF-I, oxytocin, and prostaglandin E2 output, and reduced apoptosis. Furthermore, miR-34a mimics supported effect of FSH on viability, apoptosis, progesterone, and IGF-I and inverted FSH action on proliferation, testosterone, and estradiol output. Our observations suggest that miR-34a can be involved in control of basic ovarian functions and that miR-34a and FSH are synergists in their actions on ovarian cell functions. Ability of FSH to promote miR-34a expression and ability of miR-34a mimics to increase occurrence of FSHR and to modify FSH effects suggest the existence of self-stimulating FSH-miR-34a axis, and that miR-34a can mediate actions of FSH on ovarian cells.

High carbohydrate increases amylase, plasma glucose, and gene expression related to glycolysis in giant gourami Osphronemus goramy.

The present study elucidated hepatic molecular and physiological responses of giant gourami to high dietary carbohydrates. Two levels of dietary carbohydrate, normal carbohydrate (34%, NC) and high carbohydrate (53%, HC), were offered to the fish for 60days. We evaluated the expression of genes that are related to carbohydrate metabolism, lipogenic capacities, amino acid catabolism, Krebs cycle, and energy sensing. In addition, we also observed the digestive enzyme activities, plasma glucose, glycogen content, whole-body composition, and growth performance of the fish. On day 30 after treatment, fish fed with high dietary carbohydrate level has significantly higher expression of gck, pk, hk, and ldh than the NC group (P < 0.05). In contrast, fish in the HC group had lower expression of irs1, igf-1, sdh, fbp, g6pc, gyp, and ampk compared to the NC fish (P < 0.05). On day 60 of the feeding trial, gck and hk expressions were still higher in the HC group (P < 0.05), and gyp, gdh, and ampk became increasingly expressed in the HC group. The increase of dietary carbohydrates resulted in significant increases in amylase and protease activity, plasma glucose, liver glycogen, crude protein, and lipid contents of the fish whole-body (P < 0.05). The high carbohydrate feeding reduced the fish growth rate but increased feed efficiency and did not affect mortality. In conclusion, giant gourami could utilize high carbohydrates due to a high amylase secretion, high modulation of carbohydrate metabolism, and large glucose storage capacity.