Adiponectin Expression Research Articles

Regulatory Effect of Isomaltodextrin on a High-Fat Diet Mouse Model with LPS-Induced Low-Grade Chronic Inflammation.

This study aimed to identify the effects of isomaltodextrin (IMD) on sustaining the gut integrity and microbiota composition in a high-fat diet (HFD) with a lipopolysaccharide (LPS)-induced low-grade inflammation mouse model. The homeostasis of the immune response is important to reduce the risk of developing metabolic syndromes. The results of this study showed that pre-treatment of IMD at 5% (w/v) suppressed the concentration of endotoxin and pro-inflammatory mediators TNF-α, MCP-1, and IL-6 while increasing the adiponectin level in the plasma. Subsequently, IMD supplementation maintained the structural integrity and intestinal permeability by upregulating the tight junction protein expressions, leading to reducing D-mannitol concentration in the blood. In addition, dysbiosis was observed in mice induced by HFD plus LPS, suggesting that unhealthy dietary factors elicit metabolic endotoxemia and associated dysbiosis to impair the barrier function. However, IMD supplementation was shown to restore the microbial diversity, promote the growth of Bacteroides-Prevotella, and upregulate the related d-glucarate and d-galactarate degradation pathways, together demonstrating the benefits of IMD as a prebiotic able to promote energy homeostasis. Our results also showed that the blood lipid profile and glucose level in the low-grade inflammation mouse model were modulated by IMD. Moreover, IMD supplementation effectively prevented the metabolic disorder and modulated immune responses in inflamed white adipose tissues by inhibiting the macrophage infiltration and restoring the adiponectin, PPAR-γ, and IRS-1 expression. These findings provide strong evidence for IMD to be a potential prebiotic that acts to sustain a healthy gut microbiota composition and barrier function. By protecting against an unhealthy diet-impaired metabolic balance and maintaining immune homeostasis, IMD may affect the development of metabolic disorders.

Aerobic exercise regulates FGF21 and NLRP3 inflammasome-mediated pyroptosis and inhibits atherosclerosis in mice.

Fibroblast growth factor 21 (FGF21), a known risk factor for atherosclerosis, is readily regulated by exercise, and it can inhibit NOD-like receptor protein 3 (NLRP3)-mediated pyroptosis. However, it is not clear whether aerobic exercise inhibits atherosclerosis via these pathways. Eight-week-old apolipoprotein E-deficient (ApoE-/-) mice on a high-fat diet were randomly divided into 1-h post-exercise (EX-1h), 24-h post-exercise (EX-24h), and sedentary (SED) groups. C57BL/6J wild-type mice fed normal chow served as controls (WT group). Mice in the EX-1h and EX-24h groups were subjected to treadmill exercise training for 12 weeks. Aerobic exercise reduced body weight; blood glucose, lipid, and inflammation levels; and aortic plaque area proportion. Aerobic exercise increased the sensitivity of FGF21 by upregulating the expression of the downstream receptor adiponectin (ApN); the serum FGF21 level after exercise increased initially, and then decreased. Aerobic exercise downregulated the expression of NLRP3 inflammasome-mediated pyroptosis-related markers in the aorta, and FGF21 may participate in the above process. Meanwhile, the liver may be the tissue source of serum FGF21 during aerobic exercise. In conclusion, aerobic exercise may inhibit atherogenesis by regulating FGF21 and NLRP3 inflammasome-mediated pyroptosis. Our study provides new information on the atherosclerosis-preventing mechanism of aerobic exercise.

Beneficial effects of UDCA and norUDCA in a rodent model of steatosis are linked to modulation of GPBAR1/FXR signaling

Non-alcoholic steatosis (NAFLD) and steatohepatitis (NASH) are two highly prevalent human disorders for which therapy remains suboptimal. Bile acids play an essential role in regulating liver metabolism, and several bile acids-based therapy are currently investigated for their potential therapeutic efficacy in NAFLD/NASH. Bile acids exert their functions, at least in part, by modulating two main receptors the Farnesoid-x-receptor (FXR) and the G protein-coupled receptor, GPBAR1. In the present study we have compared the pharmacological effects of two bile acids, the ursodeoxycholic acid (UDCA) and its derivative norUDCA, in a model of NAFLD/NASH induced by feeding mice with a Western diet for 12 weeks. The results of these studies demonstrated that both UDCA and norUDCA protected against development of steatosis and fibrosis, but did not reduce the hepatocytes ballooning nor the development of a pro-atherogenic lipid profile. Both agents reduced liver lipogenesis and ameliorated insulin sensitivity and adipocytes signaling as shown by increased expression of adiponectin. Mechanistically, UDCA acts as weak GPBAR1 agonist, while norUDCA exerted no effect on both GPBAR1 and FXR. In vivo administration of UDCA resets bile acid synthesis and promotes a shift toward bile acids species that are GPBAR1 agonists, UDCA, TUDCA and hyodeoxycholic acid, and increases GLP1 expression in the ileum. In contrast norUDCA is poorly metabolized exerting a minimal impact on GPBAR1 signaling. Together, these data, highlight the potential role of UDCA and norUDCA in treating of NAFLD, though these beneficial effects are supported by different mechanisms.

Abdominal adipose tissue associates with adiponectin and TNF in middle-aged healthy men

Background and Aims : Backgroun: Adipokines, including adiponectin, visfatin and TNF (tumor necrosis factor)-α, are highly active bio-peptides involved in glucose metabolism, insulin regulation and the development and progression of obesity and its associated diseases. It includes, among others Aim: In this cross-sectional study, we aimed to investigate whether gene expression levels in subcutaneous adipose tissue (SAT) of selected adipokines and their corresponding circulating levels associate with the amount of AT in superficial (sSAT), deep (dSAT) and visceral (VAT) AT, assessed by computed tomography (CT). Any association with glucometabolic variables were also exploredMethods: Methods: In 103 healthy Caucasian men, fasting venous blood and SAT samples from the gluteal region were collected. Ninety-four of the participants underwent CT assessment of the abdominal AT, which was divided into VAT and superficial - and deep subcutaneous AT (sSAT and dSAT). Circulating levels were measured by ELISA and AT gene-expression by PCR. Insulin sensitivity was determined by glucose clamp, assessing glucose disposal rate (GDR).Results: Results: Circulating adiponectin and TNF gene expression correlated inversely and positively to sSAT, dSAT and VAT (r=-0.266 to -0.276, p<0.05 for all) and (r=0.323 - 0.368, p<0.05 for all), respectively, with strongest correlations to the amount of AT in sSAT and dSAT. Circulating adiponectin correlated inversely to insulin, C-peptide and waist circumference (r=-456 to -0.373, p<0.001) and positively to GDR (r=0.356, p<0.001). AT-expressed visfatin correlated inversely to insulin and C-peptide (r=-0.370 and r=-0.404, p<0.001).Conclusions: Conclusion: Increased amount of AT is associated with low levels of adiponectin and increased levels of TNF. Background and Aims : Backgroun: Adipokines, including adiponectin, visfatin and TNF (tumor necrosis factor)-α, are highly active bio-peptides involved in glucose metabolism, insulin regulation and the development and progression of obesity and its associated diseases. It includes, among others Aim: In this cross-sectional study, we aimed to investigate whether gene expression levels in subcutaneous adipose tissue (SAT) of selected adipokines and their corresponding circulating levels associate with the amount of AT in superficial (sSAT), deep (dSAT) and visceral (VAT) AT, assessed by computed tomography (CT). Any association with glucometabolic variables were also explored Methods: Methods: In 103 healthy Caucasian men, fasting venous blood and SAT samples from the gluteal region were collected. Ninety-four of the participants underwent CT assessment of the abdominal AT, which was divided into VAT and superficial - and deep subcutaneous AT (sSAT and dSAT). Circulating levels were measured by ELISA and AT gene-expression by PCR. Insulin sensitivity was determined by glucose clamp, assessing glucose disposal rate (GDR). Results: Results: Circulating adiponectin and TNF gene expression correlated inversely and positively to sSAT, dSAT and VAT (r=-0.266 to -0.276, p<0.05 for all) and (r=0.323 - 0.368, p<0.05 for all), respectively, with strongest correlations to the amount of AT in sSAT and dSAT. Circulating adiponectin correlated inversely to insulin, C-peptide and waist circumference (r=-456 to -0.373, p<0.001) and positively to GDR (r=0.356, p<0.001). AT-expressed visfatin correlated inversely to insulin and C-peptide (r=-0.370 and r=-0.404, p<0.001). Conclusions: Conclusion: Increased amount of AT is associated with low levels of adiponectin and increased levels of TNF.

Smooth Muscle Insulin Receptor Deletion Causes Voiding Dysfunction: A Mechanism for Diabetic Bladder Dysfunction.

Diabetic bladder dysfunction (DBD) is the most common complication in diabetes. Myogenic abnormalities are common in DBD; however, the underlying mechanisms leading to these remain unclear. To understand the importance of smooth muscle insulin receptor (IR)-mediated signaling in the pathogenesis of DBD, we conditionally deleted it to achieve either heterozygous (SMIR+/-) or homozygous (SMIR-/-) deletion in smooth muscle cells. Despite impaired glucose and insulin tolerance seen with SMIR-/- mice, both SMIR+/- and SMIR-/- mice exhibited normal blood glucose and plasma insulin levels. Interestingly, these mice had abnormal voiding phenotypes, that included urinary frequency and small voids, and bladder smooth muscle (BSM) had significantly diminished contraction force. Morphology revealed a dilated bladder with thinner BSM layer, and BSM bundles were disorganized with penetrating interstitial tissue. Deletion of IR elevated FoxO and decreased mTOR protein expression, which further decreased the expression of Chrm3, P2x1, Sm22, and Cav1.2, crucial functional proteins for BSM contraction. Furthermore, we determined the expression of adiponectin in BSM, and deletion of IR in BSM inhibited adiponectin-mediated signaling. In summary, disruption of IR-mediated signaling in BSM caused abnormalities in proliferation and differentiation, leading to diminished BSM contractility and a voiding dysfunction phenotype that recapitulates human DBD.

Electroacupuncture improves follicular development and metabolism and regulates the expression of adiponectin, AMPK and ACC in an obese rat model of polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is a common endocrine disorder characterized by hyperandrogenism and follicular arrest. Electroacupuncture (EA) has been shown to be effective at improving hyperandrogenism and follicular arrest in PCOS; however, its mechanism of action remains to be deciphered. In this study, we investigated whether EA improved follicular development in an obese rat model of PCOS and regulated the expression of adiponectin, AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC). EA was administered at CV3, CV4 and ST40. Changes in body weight, paraovarian fat, estrus cycle, ovarian morphology, levels of related hormones, and glucose and lipid metabolism were evaluated. In addition, protein and mRNA expression of adiponectin, AMPK and ACC was measured. The body weight and paraovarian fat of rats in the EA group were reduced, while estrus cyclicity and ovarian morphology improved. Levels of free fatty acids, triglycerides, total cholesterol and low-density lipoprotein cholesterol were significantly reduced in the EA group, as well as blood glucose levels. Furthermore, levels of testosterone and luteinizing hormone were reduced in the EA group, while estradiol levels were increased. Protein and mRNA expression of adiponectin, AMPKα1 and liver kinase B1 (LKB1) was found to be increased in the EA group, while protein and mRNA expression of ACC were significantly reduced.Conclusion: Our findings suggest that EA improved follicular development and metabolism and regulated expression levels of adiponectin, AMPKα1, LKB1 and ACC in our obese rat model of PCOS.

Reverting to a Healthy Diet during Lactation Normalizes Maternal Milk Lipid Content of Diet-Induced Obese Rats and Prevents Early Alterations in the Plasma Lipidome of the Offspring.

ScopeThis study aims to assess in rats whether normalizing maternal diet during lactation prevents the harmful effects of western diet (WD) consumption during the whole perinatal period on the lipidomic profile in maternal milk and offspring plasma.Methods and ResultsControl dams (CON‐dams), fed with standard diet (SD); WD‐dams, fed with WD prior and during gestation and lactation; and reversion dams (REV‐dams), fed as WD‐dams but moved to SD during lactation are followed. Lipidomic analysis is performed in milk and plasma samples from pups. Milk of WD‐dams presents a different triacylglycerol composition and free fatty acid (FA) profile compared to CON‐dams, including an increased ratio of pro‐inflammatory to anti‐inflammatory long‐chain polyunsaturated FA. Such alterations, which are also present in the plasma of their offspring, are widely reversed in the milk of REV‐dams and the plasma of their pups. This is related with the recovery of control adiponectin expression levels in the mammary gland, and the presence of decreased expression of pro‐inflammatory factors.ConclusionImplementing a healthy diet during lactation prevents early alterations in the plasma lipidome of pups associated to the maternal intake of an obesogenic diet, which may be related to the normalization of milk lipid content and the inflammatory state in the mammary gland.

Different Seasonal Collections of Ficus carica L. Leaves Diversely Modulate Lipid Metabolism and Adipogenesis in 3T3-L1 Adipocytes.

Due to the high prevalence of obesity and type 2 diabetes, adipogenesis dysfunction and metabolic disorders are common features in the elderly population. Thus, the identification of novel compounds with anti-adipogenic and lipolytic effects is highly desirable to reduce diabetes complications. Plants represent an important source of bioactive compounds. To date, the antidiabetic potential of several traditional plants has been reported, among which Ficus carica L. is one of the most promising. Considering that plant metabolome changes in response to a number of factors including seasonality, the aim of this study was to evaluate whether Ficus carica leaves extracts collected in autumn (FCa) and spring (FCs) differently modulate lipid metabolism and adipogenesis in 3T3-L1 adipocytes. The 1H-NMR profile of the extracts showed that FCs have a higher content of caffeic acid derivatives, glucose, and sucrose than FCa. In contrast, FCa showed a higher concentration of malic acid and furanocoumarins, identified as psoralen and bergapten. In vitro testing showed that only FCa treatments were able to significantly decrease the lipid content (Ctrl vs. FCa 25 μg/mL, 50 μg/mL and 80 μg/mL; p < 0.05, p < 0.01 and p < 0.001, respectively). Furthermore, FCa treatments were able to downregulate the transcriptional pathway of adipogenesis and insulin sensitivity in 3T3-L1 adipocytes. In more detail, FCa 80 μg/mL significantly decreased the gene expression of PPARγ (p < 0.05), C/EBPα (p < 0.05), Leptin (p < 0.0001), adiponectin (p < 0.05) and GLUT4 (p < 0.01). In conclusion, this study further supports an in-depth investigation of F. carica leaves extracts as a promising source of active compounds useful for targeting obesity and diabetes.

Abdominal Adipose Tissue Associates With Adiponectin and TNFα in Middle-Aged Healthy Men.

IntroductionAdipokines are highly active biopeptides involved in glucose metabolism, insulin regulation and the development and progression of obesity and its associated diseases. It includes, among others, adiponectin, visfatin and tumor necrosis factor alpha (TNFα). The sources of adipokines and their associations with glucometabolic variables are not completely understood.AimIn this cross-sectional study, we aimed to investigate whether gene expression levels in subcutaneous adipose tissue (SAT) of selected adipokines and their corresponding circulating levels associate with the amount of AT in superficial (sSAT), deep (dSAT) and visceral AT (VAT), assessed by computed tomography (CT). Any association with glucometabolic variables were also explored.MethodsIn 103 healthy Caucasian men, aged 39.5 years, fasting venous blood and SAT samples from the gluteal region were collected. Ninety-four of the participants underwent CT assessment of the abdominal AT, which was divided into VAT, sSAT and dSAT. Circulating levels of adipokines were measured by ELISA and AT gene-expression by PCR. Insulin sensitivity was determined by glucose clamp, assessing glucose disposal rate (GDR).ResultsCirculating adiponectin and TNFα gene expression correlated inversely and positively to the amount of AT in all three compartments (r=-0.266 to -0.276, p<0.05 for all) and (r=0.323 - 0.368, p<0.05 for all), respectively, with strongest correlations to the amount in sSAT and dSAT. When dividing AT compartments into quartiles, a tendency was observed towards lower circulating adiponectin and higher TNFα gene expression levels, respectively, with increasing amount of sSAT and dSAT. Circulating adiponectin correlated inversely to insulin, C-peptide and waist circumference (r=-456 to -0.373, p<0.001) and positively to GDR (r=0.356, p<0.001). AT-expressed visfatin correlated inversely to insulin and C-peptide (r=-0.370 and r=-0.404, p<0.001).ConclusionIncreased amount of AT is associated with lower levels of adiponectin and increased levels of TNFα AT expression.

Evaluation of serum levels of IL-6 and adiponectin in COVID-19 patients and their relationship with disease severity

Background. The SARS-CoV-2 pandemic has prompted researchers around the world to identify risk factors associated with disease severity and mortality. Results suggest that COVID-19 mortality might be due to a cytokine storm involving IL-6, and that obesity can be considered a risk factor for COVID-19 prevalence, severity, and mortality. The current study aimed to evaluate the serum levels of IL-6 and adiponectin in patients and their relationship with disease progression. Materials and methods. ELISA was used to assess the levels of IL-6 and adiponectin in serum samples from a control group and from patients with COVID-19 at the time of admission to ICU or non-ICU wards. The results were analyzed using the MannWhitney and Spearman tests. Results. Mean serum levels of adiponectin in patients admitted to ICU (10.1815.4 ng/ml) were significantly higher than patients admitted to non-ICU wards (3.143 ng/ml, p = 0.001). Mean serum IL-6 levels showed a similar pattern, however the difference was not statistically significant (p = 0.18). In addition, a significant direct correlation was observed between adiponectin expression and IL-6 (R = 0.2, p = 0.03). Conclusion. The results of this study show that serum levels of adiponectin in COVID-19 patients with severe lung involvement were significantly higher than those with less lung involvement. This finding is of high importance mainly due to the critical role of the lungs in adiponectin signaling, and as a result, adiponectin disorders may be associated with pulmonary complications in COVID-19 patients.

Poster 106: Commonly Used Orthopaedic Clinical Therapeutics Affect the Cytokine Activity of Culture-Expanded Mesenchymal Stem Cells

Objectives:Hyaluronic acid (HA), platelet-rich plasma (PRP), and methylprednisolone are widely used for the non-operative management of osteoarthritis (OA). More recently, human bone marrow derived mesenchymal stem cells (BM-MSCs) have garnered significant interest as a non-operative means for symptom modification of OA. Anti-inflammatory cytokine secretion is a major component of proposed BM-MSC action. We sought to describe the effect of HA, PRP, and methylprednisolone on the cytokine profile of BM-MSCs at multiple stages of culture expansion.Methods:Following Institutional Review Board and United States Food and Drug Administration approval, nine BM-MSC cell lines from 4 human donors underwent three stages of culture expansion: passage 2 (P2), passage 3 (P3), and passage 4 (P4). Levels of OA-related cytokines (IL-1β, IL-6, IL-8, IL-10, Stem cell Factor [SCF], Stem Cell Derived Factor-alpha [SDF-α]) were measured using Luminex multiplexing technology. Each of the BM-MSC preparations were evaluated at each passage, 24 hours following exposure to HA, PRP, or methylprednisolone. Paired T-tests and Wilcoxon signed-rank testing were performed as appropriate following assessment of normality. Significance was set at p<0.05.Results:Culture expansion significantly altered cytokine production, however treatment with current OA therapeutics further altered these responses implicating the potential of combined treatment of culture expanded hMSCs and OA therapeutics. Figure 1 demonstrates the behavior of selected cytokines after exposure to HA, PRP or methylprednisolone preparations. Further, we tested different concentrations of each of the therapeutics to determine optimal dosing. Exposure to increasing doses of HA led to reductions in BM-MSC expression of SCF, SDF-α, VEGF, CCL20, and adiponectin (p<0.05 for all). PRP increased IL-1 levels in P2 and P3 (p<0.05 for both), IL-6 in P2 (100 pg/mL vs 5500 pg/mL, p<0.001), and IL-8 in P3 (900 pg/mL vs 3900 pg/mL, p<0.001). PRP did not affect IL-10 expression (p>0.05). Aside from an increase in IL-6 production for P2 (200 pg/mL vs 1500 pg/mL, p<0.05), methylprednisolone did not have significant effects on cytokine expression.Conclusions:Orthopaedic therapeutic adjuvants influence the inflammatory cytokine profile of BM-MSCs at multiple stages of culture expansion. Although further study is needed, these therapeutics have potential to be used in tandem with BM-MSCs to create an optimal cytokine environment for treatment of degenerative joint disease.Figure 1.hMSCs and Osteoarthritis Adjuvants. Human bone marrow derived hMSCs were culture expanded over 4 passages and treated with hyaluronan (HA), platelet rich plasma (PRP) and methylprednisolone (DEPO). Culture expansion significantly altered cytokine production, however treatment with current OA therapeutics further altered these responses implicating the potential of combined treatment of culture expanded hMSCs with OA therapeutics.

Adipose and Skeletal Muscle Expression of Adiponectin and Liver Receptor Homolog-1 With Weight Loss and Aerobic Exercise.

Adiponectin is an adipokine mainly secreted by adipocytes that regulates the metabolism of lipids and glucose. Liver receptor homolog-1 (LRH-1), also named NR5A2, is a nuclear receptor that regulates lipid metabolism and homeostasis. The purpose of this study was to compare adiponectin and LRH-1 messenger RNA (mRNA) expression in adipose tissue and LRH-1 expression in skeletal muscle between men and women at baseline and to study the effects of aerobic exercise (AEX) training or weight loss (WL) on their expression. This hospital and university setting study included 62 overweight and obese men (n = 23) and women (n = 39) older than 45 years, of whom 41 completed 6 months of WL (n = 21) or AEX (n = 20). Outcomes included abdominal and gluteal adipose tissue and skeletal muscle gene expression. Adiponectin and LRH-1 mRNA expression in adipose tissue and LRH-1 mRNA expression in skeletal muscle is higher in women than in men (P < .05). Adiponectin mRNA expression in gluteal and abdominal adipose tissue did not change significantly after AEX or WL. LRH-1 mRNA expression increased both in adipose tissue and skeletal muscle after AEX (P < .05) and the change in muscle LRH-1 was different between the groups (P < .05). Adiponectin was positively correlated to LRH-1 in adipose tissue (P < .001). The change in maximal oxygen consumption related to the change in LRH-1 mRNA (r = 0.43; P = .01). LRH-1, as a nuclear reporter, may activate adiponectin mRNA expression in adipose tissue and increases after AEX.

A Signature of Exaggerated Adipose Tissue Dysfunction in Type 2 Diabetes Is Linked to Low Plasma Adiponectin and Increased Transcriptional Activation of Proteasomal Degradation in Muscle.

Insulin resistance in skeletal muscle in type 2 diabetes (T2D) is characterized by more pronounced metabolic and molecular defects than in obesity per se. There is increasing evidence that adipose tissue dysfunction contributes to obesity-induced insulin resistance in skeletal muscle. Here, we used an unbiased approach to examine if adipose tissue dysfunction is exaggerated in T2D and linked to diabetes-related mechanisms of insulin resistance in skeletal muscle. Transcriptional profiling and biological pathways analysis were performed in subcutaneous adipose tissue (SAT) and skeletal muscle biopsies from 17 patients with T2D and 19 glucose-tolerant, age and weight-matched obese controls. Findings were validated by qRT-PCR and western blotting of selected genes and proteins. Patients with T2D were more insulin resistant and had lower plasma adiponectin than obese controls. Transcriptional profiling showed downregulation of genes involved in mitochondrial oxidative phosphorylation and the tricarboxylic-acid cycle and increased expression of extracellular matrix (ECM) genes in SAT in T2D, whereas genes involved in proteasomal degradation were upregulated in the skeletal muscle in T2D. qRT-PCR confirmed most of these findings and showed lower expression of adiponectin in SAT and higher expression of myostatin in muscle in T2D. Interestingly, muscle expression of proteasomal genes correlated positively with SAT expression of ECM genes but inversely with the expression of ADIPOQ in SAT and plasma adiponectin. Protein content of proteasomal subunits and major ubiquitin ligases were unaltered in the skeletal muscle of patients with T2D. A transcriptional signature of exaggerated adipose tissue dysfunction in T2D, compared with obesity alone, is linked to low plasma adiponectin and increased transcriptional activation of proteasomal degradation in skeletal muscle.

Molecular cloning, expression and appetite regulation function of adiponectin in Siberian sturgeon (Acipenser baerii)

Adiponectin (AdipoQ) as an adipocytokine has the potential to regulate feeding behavior, but the information about adipoq in fish is limited. In this study, Siberian sturgeon adiponectin (Ssadipoq) gene was cloned encoding 264 amino acids. The amino acid identity of SsAdipoQ was low compared with that of mammals, birds, amphibians and teleost fishes. The expression of Ssadipoq in the hypothalamus was significantly decreased at 1 h and 3 h post feeding, and increased after 15-day fasting. The mature domain of AdipoQ (fAd) was inserted into expression vector pET32a and successfully expressed in Escherichia coli BL21 (DE3) after stimulated by isopropyl-β-d-thiogalactoside. Food intake at 1 h and 3 h post treatment with SsfAd protein decreased significantly (P < 0.05). The mRNA expression of pyy and cck in the valvula intestine was promoted and hypothalamic npy, agrp and pomc mRNA expression were inhibited after treatment with SsfAd protein. Furthermore, hypothalamic ampk subunits expression was associated with peripheral SsfAd treatment. In summary, present study indicate that SsfAd plays an important role in the regulation of food intake and appetite signals in Siberian sturgeon, which provides a basis for further study application of prokaryotic AdipoQ in feeding behavior regulation.

Adipocytes contribute to tumor progression and invasion of peritoneal metastasis by interacting with gastric cancer cells as cancer associated fibroblasts.

Peritoneal metastasis (PM) is one of the most common causes of noncurative surgery and the most frequent recurrence pattern in gastric cancer (GC). During the process of PM, GC cells detached from primary tumor interact with human peritoneal mesothelial cells (HPMC) overlapped with adipose tissues such as the omentum or mesentery. Although the interaction with HPMC promotes the malignancy of GC, the role of adipose tissues remains unclear. We aimed to clarify how adipose tissue are affected by adjacent primary tumors during the expression of adipokines and to elucidate whether GC cells transform adipocytes into CAFs in vitro. In addition, we investigated whether GC cells are affected by adipocytes in their ability to infiltrate. We investigated the phenotypic conversion of adipocytes during the malignant process of GC cells in vivo and in vitro. We evaluated the expression levels of adiponectin in the omental adipose tissue of gastric cancer patients by western blotting. Following adipocytes/gastric cancer cells coculture, adipocyte markers, adiponectin receptors, and inflammatory cytokine markers were detected by real-time PCR and/or western blotting in the single-cultured and co-cultured adipocytes; cancer-associated fibroblast (CAF) markers were detected by immunofluorescence and western blotting in the single-cultured and co-cultured adipocytes; invasion assays were performed in single cultured and co-cultured MKN45 and OCUM. In omental adipose tissues that are situated close to the primary tumors, the expression of adiponectin tended to decrease in patients with subserosal or serosal invasion. By co-culturing with GC cells, adipocytes were dedifferentiated and the expression levels of CAF marker FSP1 and inflammatory cytokines, PAI-1 and IL-6, significantly increased (p < 0.05). Furthermore, GC cells co-cultured with adipocytes showed enhanced invasion ability. Our findings suggest that the phenotypic conversion of adipocytes may promote the malignancy of GC in the construction of the cancer microenvironment of PM.

PDIA4, a novel ER stress chaperone, modulates adiponectin expression and inflammation in adipose tissue.

Increasing evidence supporting a causal link between obesity and endoplasmic reticulum (ER) stress in adipose tissue is being reported. Protein disulfide isomerase 4 (PDIA4) is a novel ER chaperone involved in the pancreatic β-cells pathogenesis in diabetes. However, the role of PDIA4 in obesity progression remains poorly understood. To assess the relationship between PDIA4, adiponectin, and metformin, we used the palmitate-induced inflammation in hypertrophic adipocytes and the high-fat diet-induced obesity mouse model. Our results revealed that palmitate-induced hypertrophic adipocytes exhibit obesity-associated conditions such as increased lipid accumulation, inflammation, and reduced glucose uptake. Pharmacological and genetic inhibition of PDIA4 significantly reverses these obesity-associated conditions in adipocytes. PDIA4 mechanistically promotes obesity progression via adiponectin downregulation. Furthermore, metformin modulates PDIA4 and adiponectin expression and improves obesity-associated conditions in both in vitro adipocytes and in vivo mouse models. Serum PDIA4 concentrations are also associated with body mass index, adiponectin, triglycerides, and inflammatory cytokines in humans. This is the first study demonstrating that PDIA4 modulates adipocytes by downregulating adiponectin. Moreover, metformin may serve as a potential therapeutic for preventing obesity via PDIA4-targeting.

Paradoxical Influences of Dietary Obesity in the Early Life on the Mammary Microenvironment Towards Breast Tumorigenesis in Mice

ObjectivesObesity has been shown complex associations with breast cancer: a clear positive association in post-menopausal women but an undefined negative association in pre-menopausal women. Moreover, early-life exposure plays a critical role in cancer development later in life. This study aimed to investigate how dietary obesity in early life, mimicking human childhood and adolescence, influences the mammary microenvironment towards breast tumorigenesis in mice MethodsThirty-two female C57BL/6 mice were fed a low-fat diet (LF, 10% kcal fat) or a high-fat diet (HF, 60% kcal fat) starting at 4 wks of age for 8 wks. Half mice were sacrificed after 8-wk feeding, and the rest were switched to a normal chow diet (LabDiet#5P76) for additional 12 wks. A panel of metabolic parameters, inflammatory cytokines, folate receptors, as well as genes in the tumorigenic Wnt pathway in the mammary tissue were assessed. ResultsHF (8-wk feeding) elevated body weight (p < 0.05), and the increase remains significant after switching to the normal chow diet until sacrifice at 24 wks of age. HF reduced expressions of metabolic parameters Pparγ (p = 0.08) and adiponectin (p < 0.05) and the reductions remain after switching to normal chow diet (p < 0.01). Increased expressions of some hormone-related regulators (e.g., aromatase and estrogen receptors) and pro-inflammatory cytokines (e.g., Tnf-α and Il-6) by HF were not observed until switching to the normal chow diet for 12 wks (p < 0.05). HF increased expressions of folate transporter 1 (p = 0.07) and folate receptor1 (Flor1, p < 0.01) and the significance also remains after switching to normal chow diet (p < 0.01). Paradoxical changes of Wnt-signaling target genes (Cyclin D1, C-Myc, Axin 2) and level of active β-catenin was observed: the expressions increased right after 8-wk HF feeding but decreased after switching to normal chow diet for 12 weeks (p < 0.05). ConclusionsThese interesting findings provided an important mechanistic understanding of the apparent paradoxical associations between obesity and breast tumorigenesis: early-life obesity alters mammary metabolic and inflammatory microenvironment in favor of breast tumorigenesis, whereas improves folate status towards preventing breast tumorigenesis in the later life potentially via an epigenetic mechanism. Funding SourcesThis project was supported by USDA/NIFA (Hatch#1,013,548).

1351-P: Molecular Pathogenesis of NASH through the Dysregulation of Metabolic Organ Network in the NASH-HCC Model Mouse Treated with Streptozotocin, High-Fat Diet

Introduction: Nonalcoholic steatohepatitis (NASH) is a chronic liver disease, often associated with type II diabetes, which sometimes progresses to hepatocellular carcinoma (HCC) . NASH has become a health problem worldwide, but therapeutic agents for NASH have not yet been approved. Although the mechanism of NASH progression has not been fully elucidated, the multiple parallel hits hypothesis suggests abnormalities in adipocytokines, endotoxins, etc could contribute to the development of NASH. The STAM™ model is a clinically-correlated murine NASH model which shows the same pathological progression as human patients. Diabetes is a well-known comorbidity of NASH, and it is evident in the STAM™ model but analysis has focused mostly on the liver. Considering organ crosstalk interactions between the liver, and the gut and adipose tissue in the model should be clarified. Methods: NASH was induced by injection of 200 µg streptozotocin solution 2 days after birth and feeding with HFD after 4 weeks of age. The mice were sacrificed at NASH stage. Colon samples were snap frozen in liquid nitrogen and stored at -80°C for tight junction-related protein analysis. Adipose tissue was prepared into paraffin blocks for HE staining. Blood adiponectin was analyzed to confirm changes in adipocytokine profile. Results: Tight junction-related proteins in the intestine of STAM™ showed that expression of ZO-1 decreased with the progression of disease. Increased expression of endotoxin in the blood and decreased expression of adiponectin were also observed. HE staining of adipose tissue revealed hypertrophy of adipocytes. Conclusion: The data suggested the occurrence of leaky gut, and abnormalities in adipocytokine secretion. Together with the liver, phenotypes in these organs are highly similar to human NASH patients, and might be involved in the pathogenesis of NASH. Disclosure L.P.Bui: None. Y.Sakakibara: None. R.Tanaka: None. E.H.Pigney: None. T.Hashiguchi: None.

1409-P: Revealing Heterogeneity in Human White Adipose Tissue at Single-Cell Level

White adipose tissue is characterized by substantial functional heterogeneity based on anatomic location and developmental lineages. Such differences may contribute to depot-dependent differences in adipose regulation of systemic metabolism and the relative impact of adipose distribution (e.g. subcutaneous, SQ, vs. intraabdominal, IA) on risk for type 2 diabetes. While mouse adipose depot heterogeneity at the single-cell level has recently been explored, comparative analysis of human white fat depots is still in its infancy.Hereby, we establish a single-nucleus RNA sequencing (snRNA Seq) method for isolating and sequencing intact single nuclei from frozen human WAT biopsies. Using this approach, we analyzed the transcriptome of individual nuclei from 22 WAT SQ or IA biopsies (SQ, 12 IA, 3 paired) acquired from subjects (16 females, 3 males) with or without type 2 diabetes (T2D) across a wide BMI range (Mean+/-SD = 45.4+/-10.3 kg/m2. Range: 23.6 - 60.9 kg/m2) . We show that both SQ and IA human fat depots demonstrate substantial cell-to-cell heterogeneity, and identify multiple adipose and non-adipose-related cell types resident in both tissues. Our analysis identifies several novel subclusters, including two distinct mature adipocyte subclusters, characterized by low vs. high adiponectin expression and thus potentially contributing to adipose-mediated effects on systemic metabolism. Moreover, we identify a novel cluster present in both SQ and IA depots, with a unique transcriptomic signature marked by DAPK1 and enrichment for hedgehog signaling and endocytosis KEGG terms. Comprehensive cross-sample integrative bioinformatics analysis using the software tools scVI and VISION reveals distinct molecular markers and transcriptomic signatures between depots, between clusters, and across a range of BMI, that may serve as promising targets for understanding the mechanism by which specific adipose-resident cell populations mediate metabolic risk associated with adiposity. Disclosure V.Efthymiou: None. S.Kodani: None. A.Gupta: None. F.Shamsi: None. A.Streets: None. Y.Tseng: Consultant; Cellarity. M.Patti: Consultant; AstraZeneca, Eiger BioPharmaceuticals, Hanmi Pharm. Co., Ltd., MBX Biosciences, Poxel SA, Other Relationship; Fractyl Health, Inc., Xeris Pharmaceuticals, Inc., Research Support; Dexcom, Inc. Funding Chan Zuckerberg Initiative (CZI)

1382-P: Novel Mechanisms to Reconvert Glucocorticoids into Pro-Metabolic Agents through Adiponectin

Adiponectin is the most abundant adipokine in our circulation and regulates inter-tissue communication like the adipose-muscle axis. Pharmacological modulation of this crosstalk is beneficial in promoting exercise tolerance in metabolic stress conditions like obesity. Endogenous glucocorticoids regulate metabolic function and stress response, but chronic daily intake of exogenous glucocorticoids produces or exacerbates metabolic stress. However, in stark contrast to daily intake, we discovered that intermittent regimens of glucocorticoids, e.g. once-weekly, improves exercise tolerance. However, the mechanisms underlying the differential regimen-specific effects of glucocorticoids in obesity remain unknown. Here we investigated the mechanisms differentiating once-weekly versus once-daily regimens of the same glucocorticoid prednisone in obesity. In mice with diet-induced obesity, once-weekly prednisone stimulated total and high-molecular weight adiponectin levels, improving exercise tolerance and energy expenditure. These effects were dependent upon adiponectin and were blocked by genetic ablation of the adipokine. Upregulation of Adipoq occurred through the adipose-specific glucocorticoid receptor (GR) , as this effect was blocked in mice with inducible GR ablation in adipose tissue. In contrast, once-daily prednisone decreased adiponectin levels through upregulation of known repressors of adiponectin expression in adipose tissue. The drug regimens remodeled in opposite ways also transcriptome and function in skeletal muscle after onset of obesity. Particularly, once-weekly prednisone treatment increased the muscle metabolic response to adiponectin through the CAMKK2-AMPK cascade. Our study demonstrates that intermittent glucocorticoids produce healthful metabolic remodeling in diet-induced obesity through direct upregulation of adiponectin expression in adipose tissue and adiponectin-dependent signaling in skeletal muscle. Disclosure M.Wintzinger: Employee; Cincinnati Children's Hospital. K.Miz: None. M.Panta: None. H.Durumutla: None. A.Pragasam: None. M.Quattrocelli: None. Funding National Institutes of Health grant DK121875National Institutes of Health grant HL158531CCHMC Trustee AwardCCHMC Heart Institute Translational Grant