Growth Hormone Regulation Research Articles

The Genome-Wide Identification, Characterization, and Expression Patterns of the Auxin-Responsive PbGH3 Gene Family Reveal Its Crucial Role in Organ Development

The regulation of vital plant activities by hormones is governed by a family of macromolecular peptides referred to as GH3 genes. This work analyzed the expression patterns of GH3 family genes in pear tissues using transcriptome data and bioinformatics analysis. In the Bai Li pear genome, a total of 18 PbGH3 genes were identified. Comparative evolutionary studies have shown a strong association between PbGH3 and AtGH3 class I and class II proteins. The role of PbGH3 genes in growth activities and hormone regulation was revealed using gene ontology (GO) and promoter region analysis. In addition, although certain PbGH3 genes exhibited tissue-specific expression in sepals, the majority had a ubiquitous expression across all tissues. Bioinformatics and expression studies suggest that the GH3 gene family in pears may have a role in controlling the abscission of the fruit’s sepals. This work sheds light on the pear fruit sepal shedding process and may inspire further research.

Sustainable Fish Meal-Free Diets for Gilthead Sea Bream (Sparus aurata): Integrated Biomarker Response to Assess the Effects on Growth Performance, Lipid Metabolism, Antioxidant Defense and Immunological Status.

The growth of the aquaculture industry requires more sustainable and circular economy-driven aquafeed formulas. Thus, the goal of the present study was to assess in farmed gilthead sea bream (Sparus aurata L.) how different combinations of novel and conventional fish feed ingredients supported proper animal performance in terms of growth and physiological biomarkers of blood/liver/head kidney. A 77-day feeding trial was conducted with three experimental diets (PAP, with terrestrial processed animal protein from animal by-products; NOPAP, without processed animal protein from terrestrial animal by-products; MIX, a combination of alternative ingredients of PAP and NOPAP diets) and a commercial-type formulation (CTRL), and their effects on growth performance and markers of endocrine growth regulation, lipid metabolism, antioxidant defense and inflammatory condition were assessed at circulatory and tissue level (liver, head kidney). Growth performance was similar among all dietary treatments. However, fish fed the PAP diet displayed a lower feed conversion and protein efficiency, with intermediate values in MIX-fed fish. Such gradual variation in growth performance was supported by different biomarker signatures that delineated a lower risk of oxidation and inflammatory condition in NOPAP fish, in concurrence with an enhanced hepatic lipogenesis that did not represent a risk of lipoid liver degeneration.

Differential peptide-dependent regulation of growth hormone (GH): A comparative analysis in pituitary cultures of reptiles, birds, and mammals

Growth hormone (GH) is a pituitary protein that exerts pleiotropic roles in vertebrates. The mechanisms regulating GH synthesis and secretion are finely controlled by hypothalamic neuropeptides and other factors. These processes have been considerably studied in mammals but are still poorly understood in other groups. To better understand the pituitary GH regulation during vertebrate phylogeny, we compared the effects of incubating several peptides on cultures of ex-vivo pituitary fragments obtained from representative specimens of reptiles (iguana), birds (chicken) and mammals (rat). The peptides used were: growth hormone-releasing hormone (GHRH), thyrotropin-releasing hormone (TRH), pituitary adenylate cyclase-activating polypeptide (PACAP), ghrelin, gonadotropin-releasing hormone (GnRH), and somatostatin (SST). In rat pituitary cultures, GH secretion was stimulated by GHRH and TRH, while gh mRNA expression was increased by GHRH and PACAP. In the case of chicken pituitaries, GH release was promoted by GHRH, ghrelin, PACAP, and GnRH, although the latter two had a dual effect since at a shorter incubation time they decreased GH secretion; in turn, gh mRNA expression was significantly stimulated by TRH, PACAP, and GnRH. The most intense effects were observed in iguana pituitary cultures, where GH secretion was significantly augmented by GHRH, PACAP, TRH, ghrelin, and GnRH; while gh mRNA expression was stimulated by GHRH, TRH, and PACAP, but inhibited by ghrelin and SST. Also, in the three species, SST was able to block the GHRH-stimulated GH release. Furthermore, it was found that the expression of Pou1f1 mRNA was increased with greater potency by GHRH and PACAP in the iguana, than in chicken or rat pituitary cultures. Additionally, in-silico analysis of the gh gene promoter structures in the three species showed that the reptilian promoter has more Pit-1 consensus binding sites than their avian and mammalian counterparts. Taken together, results demonstrate that pituitary peptide-mediated GH regulatory mechanisms are differentially controlled along vertebrate evolution.

Experimental Autoimmune Encephalomyelitis Influences GH-Axis in Female Rats.

Inflammation, demyelination, and axonal damage to the central nervous system (CNS) are the hallmarks of multiple sclerosis (MS) and its representative animal model, experimental autoimmune encephalomyelitis (EAE). There is scientific evidence for the involvement of growth hormone (GH) in autoimmune regulation. Previous data on the relationship between the GH/insulin like growth factor-1 (IGF-1) axis and MS/EAE are inconclusive; therefore, the aim of our study was to investigate the changes in the GH axis during acute monophasic EAE. The results show that the gene expression of Ghrh and Sst in the hypothalamus does not change, except for Npy and Agrp, while at the pituitary level the Gh, Ghrhr and Ghr genes are upregulated. Interestingly, the cell volume of somatotropic cells in the pituitary gland remains unchanged at the peak of the disease. We found elevated serum GH levels in association with low IGF-1 concentration and downregulated Ghr and Igf1r expression in the liver, indicating a condition resembling GH resistance. This is likely due to inadequate nutrient intake at the peak of the disease when inflammation in the CNS is greatest. Considering that GH secretion is finely regulated by numerous central and peripheral signals, the involvement of the GH/IGF-1 axis in MS/EAE should be thoroughly investigated for possible future therapeutic strategies, especially with a view to improving EAE disease.

Diversity of the Ghrelin Gene in Nigeria’s Fulani and Yoruba Ecotype Chickens

Ghrelin (GHRL) is a novel 28-amino acid gut-brain peptide linked to a gene associated with the regulation of growth hormones in birds. This study was carried out to investigate the polymorphism of the Ghrelin gene in Fulani and Yoruba ecotypes chickens in Nigeria. Blood samples were collected from seventy-eight (78) Nigerian indigenous chickens comprising of 41 Yoruba ecotype chickens and 37 Fulani ecotype chickens. Polymerase Chain Reaction-Restriction fragment length polymorphism (PCR-RFLP) method was used and the MboII restriction enzyme cut site 71 of the ghrelin and genetic structure were determined. Population structure was analyzed using allele and genotype frequencies, heterozygosity and genetic variation metrics. Two alleles (C and T) and three genotypes (CC, CT and TT) were observed. In the Yoruba Ecotype, the allele frequencies were C (0.34) and T (0.66) respectively while C (0.45) and T (0.55) were observed in the Fulani ecotype and the overall population was C (0.39) and T (0.61). The genotype frequencies obtained were; in the Yoruba ecotype, CC (0.10), CT (0.48), and TT (0.41) were observed. In the Fulani ecotype, CC (0.22), CT (0.45), and TT (0.32) were also observed, and in the overall population CC (0.15). CT (0.47) and TT (0.37) were observed. FIS values for the Yoruba ecotype (-0.0847) and Fulani ecotype (0.00702) reflects random mating and inbreeding respectively. The effective number of alleles indicates that the Fulani ecotype has more effective alleles compared to the Yoruba ecotype. These results suggest that the Yoruba ecotype may be at Hardy-Weinberg equilibrium, while Fulani ecotype deviates for the ghrelin locus. In summary, our results may open opportunities for genetic improvement in Nigerian indigenous chicken due to the polymorphic nature of the ghrelin gene.

The effects of nano-curcumin on growth performance, feed utilization, blood biochemistry, disease resistance, and gene expression in European seabass (Dicentrarchus labrax) fingerlings

Phytochemicals are used in fish farming to reduce stress and combat diseases during intensification. Recently, nanotechnology has represented a paradigm shift in the aquatic feed industry to improve the solubility, availability, and efficacy of phytochemicals. The purpose of this research was to investigate the effects of dietary supplementation with nano-curcumin (CUNE) on the growth, feed utilization, body composition, blood biochemistry, antioxidant status, disease resistance to Vibrio parahaemolyticus (V. parahaemolyticus), and the expression of insulin-like growth factor 1 (IGF-1), growth hormone (GH), interleukin-10 (IL-10), and interlukin-1β (IL-1β) genes in European Seabass (Dicentrarchus labrax) fingerlings. Fish (6.02±0.01) were randomly divided into four equal groups and fed varying levels of CUNE: 0 (CUNE0), 50 (CUNE1), 60 (CUNE2), and 70 mg/kg (CUNE3) of nano-curcumin for 56 days, respectively. The inclusion of dietary CUNE significantly improved the growth indices (final body weight and weight gain, and specific growth rate) and feed utilization (improved feed intake and lower FCR) of European seabass fingerlings (p<0.05). However, CUNE inclusion had no significant effect on body composition including the percentages of dry mater, crude protein, crude lipid, and ash (p>0.05). All CUNE groups established an enhancement in blood hematology in a dose-dependent manner, with CUNE3 demonstrating the highest values. CUNE supplementation (70 mg/kg diet) produced the highest levels of total protein, albumin, globulin, and triglycerides, as well as the lowest levels of glucose, and hepatic enzymes (ALT, AST, and ALP; p<0.05) compared to other groups. Furthermore, the CUNE-supplemented groups showed significant improvements in superoxide dismutase, catalase, and glutathione peroxidase, while reducing malondialdehyde (p<0.05). Additionally, the consumption of CUNE increased the expression of IGF-1, GH, and IL-10 genes in a dose-dependent manner (p<0.05). Interestingly, fish fed 70 mg of CUNE in their diets had lower expression of the IL-1β gene compared to the other groups (p<0.05). When the fish were challenged with V. parahaemolyticus, the addition of CUNE in their diets resulted in reduced mortality rates. Therefore, nano-curcumin could be considered as a natural feed additive to promote growth, feed utilization, and bacteria resistance via supporting antioxidant status, serum metabolites, and the regulation of GH, IL-10, and IGF-1 in the hepatic tissues of European seabass fingerlings.

Research Note: SOCS2 contributes to reduction of the third digit during development of the chicken forelimb

The development of the avian wing pattern has been the subject of heated debate due to its special shape. The Suppressor of cytokine signaling 2 (SOCS2) gene encodes a negative regulator of growth hormone (GH) signaling and bone growth and is known to be strongly expressed in the third digit of chicken forelimbs. These observations suggest that SOCS2 might regulate the morphology of the avian wing, however, the function of SOCS2 in avian limb development remains unknown. Here, we reexamined SOCS2 expression in successive developmental stages of chicken limb development by in situ hybridization (ISH) and describe extended expression from the posterior of the stypolod to the third digit of the forelimbs. We used the RCAS avian retrovirus to overexpress SOCS2 in the developing chicken limb buds, which resulted in reduced or malformed chicken wings while hindlimbs developed normally. Transcriptome sequencing (mRNA-Seq) revealed changes in expression of genes known to be associated with growth and development in forelimbs with overexpressed SOCS2. This study highlights a pivotal role for SOCS2 during the development of the wing in the chicken and provides new insight into molecular mechanisms regulating avian limb development.

Characterization of the somatostatin system in tilapia: implications for growth and reproduction.

Somatostatin (SST) plays diverse physiological roles in vertebrates, particularly in regulating growth hormone secretion from the pituitary. While the function of SST as a neuromodulator has been studied extensively, its role in fish and mammalian reproduction remains poorly understood. To address this gap, we investigated the involvement of the somatostatin system in the regulation of growth and reproductive hormones in tilapia. RNA sequencing of mature tilapia brain tissue revealed the presence of three SST peptides: SST6, SST3, and low levels of SST1. Four different isoforms of the somatostatin receptor (SSTR) subfamily were also identified in the tilapia genome. Phylogenetic and synteny analysis identified tiSSTR2-like as the root of the tree, forming two mega clades, with SSTR1 and SSTR4 in one and SSTR2a, SSTR3a, and SSTR5b in the other. Interestingly, the tiSSTR-5 isoforms 5x1, 5x2, and 5x3 were encoded in the sstr3b gene and were an artifact of misperception in the nomenclature in the database. RNA-seq of separated pituitary cell populations showed that SSTRs were expressed in gonadotrophs, with sstr3a enriched in luteinizing hormone (LH) cells and sstr3b significantly enriched in follicle-stimulating hormone (FSH) cells. Notably, cyclosomatostatin, an SSTR antagonist, induced cAMP activity in all SSTRs, with SSTR3a displaying the highest response, whereas octreotide, an SSTR agonist, showed a binding profile like that observed in human receptors. Binding site analysis of tiSSTRs from tilapia pituitary cells revealed the presence of canonical binding sites characteristic of peptide-binding class A G-protein-coupled receptors. Based on these findings, we explored the effect of somatostatin on gonadotropin release from the pituitary in vivo. Whereas cyclosomatostatin increased LH and FSH plasma levels at 2h post-injection, octreotide decreased FSH levels after 2h, but the LH levels remained unaffected. Overall, our findings provide important insights into the somatostatin system and its mechanisms of action, indicating a potential role in regulating growth and reproductive hormones. Further studies of the complex interplay between SST, its receptors, and reproductive hormones may advance reproductive control and management in cultured populations.

The Role of Chronic Inflammation in Stunting: A Systematic Literature Review

Chronic inflammation can overactivate the body's immune system, causing excessive release of cytokines and inflammatory substances. This can affect the balance between the inflammatory response and the immune response, so that the immune system may be more focused on fighting inflammation rather than protecting the body from infection. Chronic inflammation can disrupt the body's growth pathways. Some molecules involved in the growth process, such as growth hormone, can be disrupted by inflammation. Additionally, inflammation can affect the development of the body's organs, tissues, and cells in ways that inhibit linear growth. This study aims to explore the role of chronic inflammation in the incidence of stunting. The literature search process was carried out on various databases (PubMed, Web of Sciences, EMBASE, Cochrane Libraries, and Google Scholar) regarding the role of chronic inflammation in the incidence of stunting. This study follows the preferred reporting items for systematic reviews and meta-analysis (PRISMA) recommendations. Chronic inflammation plays an important role in the occurrence of stunting in children. Chronic inflammation can influence various aspects that contribute to the growth and development of children's bodies, including nutrient absorption, growth hormone regulation, nutrient metabolism, and immune system function. This can result in stunted child growth, malnutrition, weight loss, and negative impacts on child well-being.

Specific features of endocrine regulation of plastic processes in premature infants and children small for gestational age

Low-birthweight and preterm infants have high risk of obesity and obesity-related diseases in the future. This review article identifies risk factors and endocrine biomarkers with greatest predictive value to the metabolic diseases development. Low concentrations of IGF-1 in low-birthweight children are associated with adipogenesis. Low leptin levels may be considered as a biomarker of catch-up growth. Long term programming effects of in utero exposure to leptin extend beyond infancy into early childhood. Adiponectin levels are positively correlated with obesity in early life, but not at ages older than three years. Rapid postnatal growth rate is associated with metabolic syndrome. Conclusion. Specific features of endocrine regulation of growth and dynamics of plastic processes in premature infants and SGA children are associated with excessive accumulation of adipose tissue, which can function as a mechanism for metabolic programming of distant endocrine and cardiometabolic disorders.

Effects of Vermicompost on Soil and Plant Health and Promoting Sustainable Agriculture

This review highlights the potential of vermicompost and its derived products as sustainable and eco-friendly solutions for enhancing production and pest management in grain crops. It assesses their impact comprehensively on crops such as maize, wheat, barley, rice, and pearl millet. Vermicompost improves soil quality, increases nutrient availability, boosts crop productivity, and enhances pest and disease tolerance. It acts as an organic fertilizer, enriching the soil with essential nutrients, humic acids, growth-regulating hormones, and enzymes, improving plant nutrition, photosynthesis, and overall crop quality. Furthermore, vermicompost shows promise in mitigating soil degradation and sequestering organic carbon while demonstrating the potential for pest management, including effectiveness against pests like fall armyworm (Spodoptera frugiperda). This review emphasizes the importance of integrated nutrient management and proper application strategies to maximize the benefits of vermicompost in grain crops. Factors such as the form and timing of application, efficacy against specific pests, and economic viability for different farming scales are discussed. Understanding these factors is crucial for successfully implementing and adopting vermicompost-based pest management strategies in grain crops. This review also explores the potential of vermicomposting as an eco-friendly and cost-effective solution to remediate organic contaminants, emerging contaminants, personal-care and pharmaceutical products, and microplastics. The review further identifies knowledge gaps and highlights the need for future studies to effectively utilize vermicompost and its derived products in cereal production for sustainable agriculture, contributing to global food security.

Effects of topping and non-topping on growth-regulating hormones of flue-cured tobacco (Nicotiana tabacum L.)-a proteomic analysis.

Until now, the mechanism underlying the impact of topping on hormone regulation in tobacco plants remains unclear, and most studies investigating the hormone signaling pathways in plants rely on genes or transcriptional pathways. This study examines the regulatory mechanisms of hormones in the roots and leaves of tobacco plants with and without topping at the protein level. The results demonstrate that, compared with non-topped plants, topping leads to a decrease in the levels of IAA (auxin), ABA (abscisic acid), and GA (gibberellin) hormones in the leaves, whereas the content of the JA (jasmonic acid) hormone increases. Furthermore, in the roots, topping results in an increase in the levels of IAA, ABA, and JA hormones, along with a decrease in GA content. In the leaves, a total of 258 significantly different proteins were identified before and after topping, with 128 proteins upregulated and 130 proteins downregulated. In the roots, there were 439 proteins with significantly different quantities before and after topping, consisting of 211 upregulated proteins and 228 downregulated proteins. Notably, these proteins were closely associated with the metabolic and biosynthetic pathways of secondary metabolites, as indicated by functional categorization. When integrating the hormone changes and the proteomics results, it is evident that topping leads to increased metabolic activity and enhanced hormone synthesis in the root system. This research provides a theoretical foundation for further investigations into the regulation and signaling mechanisms of hormones at the protein level before and after topping in plants.

THU157 Epigenetic Regulation Of Genes Involved In The Signaling Cascade Of The GH Axis - IGF1 And Insulin. Implication In Children Postnatal Growth

Abstract Disclosure: L. Zoff: None. F. Garcia: None. G. Aschettino: None. G. Veneruzzo: None. M.C. Mattone: None. N. Perez Garrido: None. M. Juanes: None. N.I. Saraco: None. C. Alonso: None. G. Guercio: None. A. Belgorosky: None. M.S. Baquedano: None. Growth is influenced by genetic, nutritional, environmental, and hormonal factors, but it proceeds in a predictable pattern characterized by a constant growth deceleration between childhood and adolescence. This growth deceleration is coordinated in tissues and organs in order to maintain body proportions. Growth hormone (GH)/ insulin-like growth factor type 1 (IGF1) axis and insulin are crucial stimulators of growth, cell proliferation and metabolism. However, their levels do not change with age in a pattern that would explain the prepubertal decline in growth rate. DNA methylation represents a fundamental epigenetic mark that is associated with transcriptional repression during development. We hypothesize that the postnatal growth pattern could be orchestrated by epigenetic mechanisms. In order to analyze age related physiological changes, we evaluated promoter methylation in the GHR, IGF1R and INSR genes in peripheral blood from 40 healthy children of both sexes (females (F) n=21 and males (M) n=19), between 3 and 15 years old by using targeted deep-amplicon bisulfite sequencing on a MiSeq system. Sequencing libraries of 4 promoter CpG rich regions for GHR, 3 for IGF1R and 5 for INSR (104, 103 and 141 CpG sites, respectively) were analyzed for mean methylation values of all CpG sites using amplikyzer2 software. The results would suggest a sexual dimorphism in the epigenetic regulation of GH, IGF1 and insulin receptors gene expression. A significant negative correlation between GHR methylation and age was observed in both sexes (Multiple Spearman correlation, p&amp;lt;0.05), but in different GHR promoter regions. Specifically, a decrease in themethylation levels with age were detected in CpG-8, CpG+183 and CpG+243 in F; and in CpG+343, CpG+346, CpG+385, CpG+391, CpG+407 and CpG+487 in M. In contrast, in the IGF1R promoter region, a positive correlation was observed exclusively in F, between mean methylation levels and age in CpG-51, CpG-43, CpG-41, CpG-23, CpG-17, CpG-15, CpG-13, CpG-11, and CpG-9, p&amp;lt;0.05. Male-specific positive correlation between INSR promotermethylation values and age were found in CpG-768, CpG+10, CpG+31, CpG+87, CpG+95, CpG+163, CpG+199, CpG+206, CpG+238, CpG+700, CpG+815 and CpG+900, p&amp;lt;0.05.Taking together, our findings are in agreement with the hypothesis that age-associated DNA promoter methylation changes could be involved in the physiological growth pattern. Finally, the observed sexual dimorphism suggests age- and sex-dependent regulatory mechanisms for normal growth. Presentation: Thursday, June 15, 2023

Environmentally relevant concentrations of Mn2+ disrupts the endocrine regulation of growth in juvenile Yunlong groupers (Epinephelus moara♀×Epinephelus lanceolatus♂)

Even though manganese is a bioelement essential for metabolism, excessive manganese levels in water can be detrimental to fish development and growth. Therefore, the aim of this study was to evaluate the effects of Mn2+ (0, 0.5,1, 2, and 4 mg·L−1) exposure for 30 d on the growth performance, growth hormone/insulin-like growth factor (GH/IGF) axis, hypothalamic-pituitary-thyroid (HPT) axis, and monoaminergic neurotransmitters of Epinephelus moara♀×Epinephelus lanceolatus♂(Yunlong grouper). Compared with the control and low Mn2+concentration groups of (0.5 and 1 mg·L−1), the high concentration of Mn2+ (4 mg·L−1) significantly reduced body weight (BW), body length (BL), weight gain rate (WGR), and specific growth rate (SGR), increased the feed coefficient rate (FCR) and mortality of Yunlong groupers (P < 0.05). Further, the levels of GH and IGF, along with the expression of ghra and ghrb were significantly reduced after exposure to 2 and 4 mg·L−1 Mn2+for 30 d, whereas the expression of sst5 was significantly up-regulated after exposure to 2 and 4 mg·L−1 Mn2+for 20 and 30 days. Moreover, Mn2+exposure increased thyroid hormone (T3) and thyroid stimulating hormone (TSH) contents, accompanied by increased mRNA levels of dio1 and dio2, however, the T4 level was decreased. Finally, dopamine (DA) and serotonin (5-HT) levels significantly decreased after long-term exposure to higher concentrations of Mn2+, and the levels their metabolites changed as well, suggesting that the synthesis and metabolism of DA and 5-HT were affected. Accordingly, changes in the GH/IGF and HPT axes-related parameters may be the cause of growth inhibition in juvenile groupers under Mn2+ exposure, indicating that the relationship between endocrine disorder and growth inhibition should not be ignored.

The Exon 3-Deleted Growth Hormone Receptor (d3GHR) Polymorphism-A Favorable Backdoor Mechanism for the GHR Function.

Growth hormone (GH) is a peptide hormone that plays a crucial role in controlling growth, development, and lifespan. Molecular regulation of GH is accomplished via the GH receptor (GHR), which is the main factor influencing human development and is essential to optimal functioning of the GH/IGF-I axis. Two GHR isoforms have been studied, according to the presence (flGHR) or absence (d3GHR) of exon 3. The d3GHR isoform, which lacks exon 3 has recently been related to longevity; individuals carrying this isoform have higher receptor activity, improved signal transduction, and alterations in the treatment response and efficacy compared with those carrying the wild type (WT) isoform (flGHR). Further, studies performed in patients with acromegaly, Prader-Willi syndrome, Turner syndrome, small for gestational age (SGA), and growth hormone deficiency (GHD) suggested that the d3GHR isoform may have an impact on the relationship between GH and IGF-I levels, height, weight, BMI, and other variables. Other research, however, revealed inconsistent results, which might have been caused by confounding factors, including limited sample sizes and different experimental methods. In this review, we lay out the complexity of the GHR isoforms and provide an overview of the major pharmacogenetic research conducted on this ongoing and unresolved subject.

Alveolar epithelial cell growth hormone releasing hormone receptor in alveolar epithelial inflammation

Purpose: Growth hormone-releasing hormone (GHRH) is a 44-amino acid peptide that regulates growth hormone (GH) secretion. We hypothesized that GHRH receptor (GHRH-R) in alveolar type 2 (AT2) cells could modulate pro-inflammatory and possibly subsequent pro-fibrotic effects of lipopolysaccharide (LPS) or cytokines, such that AT2 cells could participate in lung inflammation and fibrosis. Methods: We used human alveolar type 2 (iAT2) epithelial cells derived from induced pluripotent stem cells (iPSC) to investigate how GHRH-R modulates gene and protein expression. We tested iAT2 cells’ gene expression in response to LPS or cytokines, seeking whether these mechanisms caused endogenous production of pro-inflammatory molecules or mesenchymal markers. Quantitative real-time PCR (RT-PCR) and Western blotting were used to investigate differential expression of epithelial and mesenchymal markers. Result: Incubation of iAT2 cells with LPS increased expression of IL1-β and TNF-α in addition to mesenchymal genes, including ACTA2, FN1 and COL1A1. Alveolar epithelial cell gene expression due to LPS was significantly inhibited by GHRH-R peptide antagonist MIA-602. Incubation of iAT2 cells with cytokines like those in fibrotic lungs similarly increased expression of genes for IL1-β, TNF-α, TGFβ-1, Wnt5a, smooth muscle actin, fibronectin and collagen. Expression of mesenchymal proteins, such as N-cadherin and vimentin, were also elevated after prolonged exposure to cytokines, confirming epithelial production of pro-inflammatory molecules as an important mechanism that might lead to subsequent fibrosis. Conclusion: iAT2 cells clearly expressed the GHRH-R. Exposure to LPS or cytokines increased iAT2 cell production of pro-inflammatory factors. GHRH-R antagonist MIA-602 inhibited pro-inflammatory gene expression, implicating iAT2 cell GHRH-R signaling in lung inflammation and potentially in fibrosis.

Insights on discovery, efficacy, safety and clinical applications of ghrelin receptor agonist capromorelin in veterinary medicine.

Growth hormone and insulin like growth factor-1 plays an important role in the regulation of body composition and metabolism. Growth Hormone is released from the pituitary through a specific G-protein coupled receptor (GPCR) called growth hormone secretagogue receptor 1a expressed in the hypothalamus. Ghrelin is a peptide hormone released from the cells in the stomach, which stimulates appetite and food intake in mammals, regulates gut motility, gastric acid secretion, taste sensation, circadian rhythm, learning and memory, oxidative stress, autophagy, glucose metabolism etc. When the release of the endogenous ligand GHSR-1a, i.e., ghrelin is malfunctioned or stopped, external substitutes are administrated to induce the stimulation of growth hormone and appetite. A class of compound known as ghrelin receptor agonists are developed as an external substitute of ghrelin for regulation and stimulation of growth hormone in frailty, for body weight gain, muscle mass gain, prevention of cachexia and for the treatment of chronic fatigue syndromes. Capromorelin [Entyce™ (Aratana Therapeutics, Leawood, KS, USA)] is the only FDA (Food and Drug Administration) approved (May 2016) drug used for stimulating appetite in dogs and was marketed in the fall of 2017. In 2020, USFDA approved Capromorelin [Elura™ (Elanco US Inc.)] for the management of weight loss in chronic kidney disease of cats. This article reviews the discovery of the ghrelin receptor agonist capromorelin, its efficacy, safety, clinical applications and aims to delineate its further scope of use in veterinary practice.

O-206 Growth hormone supplementation reduces aneuploidy and improves the quality of aged oocytes by activating the JAK2-ERK1/2 pathway

Abstract Study question Does growth hormone (GH) supplementation improves the quality of oocytes in aged mice via reducing aneuploidy? Summary answer Daily injections of GH for 8 weeks reduces aneuploidy by activating JAK2-ERK1/2 pathway and improves the quality of oocyte in aged mice. What is known already Aging-induced low-quality oocytes, which are mainly caused by aneuploidy, decrease female fertility. However, few treatments are available to improve the poor quality of oocytes in aged women. Age-related decline in GH levels may be related to the insufficient reproductive potential in aged women. Several studies have reported that GH improves assisted reproductive technology outcomes in poor ovarian response patients, although its role is still controversial. In animal models, GH supplementation improved the oocyte quality by enhancing mitochondrial function. However, whether GH reduces the aneuploidy rate in aged oocytes, and if so the underlying mechanisms, are unclear. Study design, size, duration The aged (8-month-old) C57BL/6J female mice were used in the study. For the in vivo experiments, the aged mice were intraperitoneally injected with GH (GenSci, Changchun, China; 1.6 mg/kg body weight) or an equivalent volume of normal saline for 8 weeks; For the in vitro experiments, germinal vesicle (GV) oocytes from aged mice were treated with GH (200 ng/mL) or PBS in M16 medium. Participants/materials, setting, methods We counted oocytes by immunohistochemistry to assess ovarian reserve. The expression of growth hormone receptor and mitochondrial genes were measured by quantitative real-time PCR. Time–lapse incubator was utilized to record the developmental potential of early embryos. Immunofluorescence was performed to assess parameters of oocyte quality (mitochondrial functions, spindle/chromosome defects, and DNA damage). Chromosome spread and DNA sequencing were used to analyze aneuploidy rate. We performed quantitative proteomics analysis to identify the potential targets of GH. Main results and the role of chance Firstly, we found that GH promoted GHR expression in aged oocytes (30.64±1.70 vs 21.68±1.08, P &amp;lt; 0.001). GH ameliorated aging-induced decline in ovarian reserve of aged mice, with increased ovarian index (0.040%±0.0031 vs 0.023%±0.0019, P &amp;lt; 0.01), and number of antral follicles (8.67±1.2 vs 3.33±0.88, P &amp;lt; 0.05) compared to the controls. In addition, GH improved the quality of aged oocytes, representing restored mitochondrial functions (P &amp;lt; 0.05), decreased DNA damage (23.37±1.76 vs 38.35±2.52, P &amp;lt; 0.001) . As expected, GH promoted the fertilization rate (39.9%±1.10 vs 14.4%±0.62, P &amp;lt; 0.001) and early embryo development (P &amp;lt; 0.01) of aged oocytes. Of note, GH recovered the spindle/chromosome defects (42.6%±2.12 vs 63.9%±1.83, P &amp;lt; 0.01, and reduced aneuploidy rate (20.9%±1.61 vs 39.3%±1.84, P &amp;lt; 0.01) in aged oocytes. GH activated ERK1/2 expression to decrease aneuploidy rate in aged oocytes. In addition, JAK2 might be involved in the regulation of GH on ERK1/2 expression in aged oocytes. Altogether, GH restored age-related decline in oocyte number, and decreased the occurrence rate of aneuploidy by activating JAK2-ERK1/2 pathway, thereby improving the quality of aged oocytes. Limitations, reasons for caution The molecular action mechanism of GH to regulate aneuploidy remains to be determined. Besides, future work should be extended to human oocyte to determine whether this mechanism is conserved between mice and humans Wider implications of the findings Our work expounds a theoretical basis for application of GH to improve the fertility of aged women. Besides, the results also feed new ideas for the prevention and treatment of oocyte quality decline in assisted reproductive technology. Trial registration number not applicable

Estradiol and progesterone regulate NUCB2/nesfatin-1 expression and function in GH3 pituitary cells and THESC endometrial cells

ABSTRACT Estradiol (E2) and progesterone (P4) are essential sex steroid hormones that play critical roles in the pituitary gland and uterus. Recently, nesfatin-1, a polypeptide hormone that regulates appetite and energy homeostasis in the hypothalamus, was found to be expressed in the pituitary gland and uterus. In this study, we aimed to investigate the relationship between these two steroid hormones and the expression and function of nesfatin-1 in the pituitary gland and uterus using GH3 cells, a lacto-somatotroph cell line, and THESC cells, an endometrial stromal cell line. First, we verified the presence of nesfatin-1 and nesfatin-1 binding sites in GH3 and THESC cells. E2 increased the mRNA expression of NUCB2, the gene encoding the nesfatin-1 protein, in GH3 cells, while P4 had no significant effect. In THESC cells, NUCB2 mRNA expression was decreased by E2 but increased by P4. In addition, nesfatin-1 significantly increased growth hormone (GH) and prolactin (PRL) mRNA expression in GH3 cells, and E2 enhanced this effect. In THESC cells, nesfatin-1 significantly increased the mRNA expression of insulin-like growth factor binding protein 1 (IGFBP1) and PRL, which are decidualization marker genes, and P4 further enhanced this effect. These results suggest that nesfatin-1 may act as a local regulator of GH and PRL production in the pituitary gland and decidualization in the uterus, modulating its effects in response to E2 and P4.

Deletion of hepatic growth hormone receptor (GHR) alters the mouse gut microbiota by affecting bile acid metabolism

ABSTRACT Both growth hormone (GH) and gut microbiota play significant roles in diverse physiological processes, but the crosstalk between them is poorly understood. Despite the regulation of GH by gut microbiota, study on GH’s influence on gut microbiota is limited, especially on the impacts of tissue specific GH signaling and their feedback effects on the host. In this study, we profiled gut microbiota and metabolome in tissue-specific GHR knockout mice in the liver (LKO) and adipose tissue (AKO). We found that GHR disruption in the liver rather than adipose tissue affected gut microbiota. It changed the abundance of Bacteroidota and Firmicutes at phylum level as well as abundance of several genera, such as Lactobacillus, Muribaculaceae, and Parasutterella, without affecting α-diversity. Moreover, the impaired liver bile acid (BA) profile in LKO mice was strongly associated with the change of gut microbiota. The BA pools and 12-OH BAs/non-12-OH BAs ratio were increased in the LKO mice, which was due to the induction of CYP8B1 by hepatic Ghr knockout. Consequently, the impaired BA pool in cecal content interacted with gut bacteria, which in turn increased the production of bacteria derived acetic acid, propionic acid, and phenylacetic acid that were possible to participate in the impaired metabolic phenotype of the LKO mice. Collectively, our findings suggested that the liver GH signaling regulates BA metabolism by its direct regulation on CYP8B1, which is an important factor influencing gut microbiota. Our study is significant in exploring gut microbiota modification effects of tissue-specific GH signaling as well as its involvement in gut microbiota–host interaction.