AdipoR1 mRNA Research Articles

Role of Adiponectin Gene and Receptor Polymorphisms and Their mRNA Levels with Serum Adiponectin Level in Myocardial Infarction.

Background and AimGenetic factors are vital participants in the development and progression of myocardial infarction (MI). Adiponectin has been assumed to have a protective role in MI and adiponectin receptors variants could be a determinant for atherosclerosis. We aimed to evaluate the prevalence of ADIPOQ (rs2241766) and ADIPOR2 (rs10773989) polymorphisms and their association with mRNA levels and circulatory adiponectin levels in patients with MI.Subjects and MethodsA total of 220 participants were classified into two groups: group 1 included 120 patients with MI, and group 2 involved 100 healthy participants as controls. Genotyping of ADIPOQ (rs2241766) and ADIPOR2 (rs10773989) polymorphisms were analyzed using an allele discrimination assay with real-time PCR and their relative expression or mRNA levels were determined by real-time PCR. Serum adiponectin level was determined using an ELISA technique.ResultsThe ADIPOQ rs2241766 GG genotype and G allele and the CC genotype and C allele of ADIPOR2 rs10773989 were significantly prevalent in patients with MI and associated with increased risk of MI. We detected a marked reduction in serum adiponectin, ADIPOQ and ADIPOR2 mRNA levels in patients than control. The GG genotype of ADIPOQ rs2241766 and the CC genotype of ADIPOR2 rs10773989 had the lowest levels of their mRNA and adiponectin level in both patients and controls.ConclusionAdiponectin gene and receptor variants are potentially related to MI risk; furthermore, their expressions were markedly depressed in MI which suggests their use as potential biomarkers for MI.

Adiponectin Signaling and Impaired GTPase Rab5 Expression in Adipocytes of Adolescents with Obesity

Introduction: Abnormalities in adipose tissue AdipoR1; adaptor protein, phosphotyrosine interaction, PH domain, and leucine zipper containing 1 (APPL1); GTPase Rab5; adiponectin; leptin; and visfatin in adults with obesity have been associated with metabolic disturbances. Objective: The objective of this study was to examine whether pediatric obesity disrupts elements of the adiponectin signaling pathway and GTPase Rab5 in adipose tissue. Methods: Primary adipocyte cultures of subcutaneous abdominal tissue were obtained from 96 lean and 66 children and adolescents with obesity (AO). AdipoR1, APPL1, and GTPase Rab5 mRNA levels were measured by RT-PCR and their protein content by Western immunoblotting. Serum total and high-molecular-weight adiponectin, leptin, leptin soluble receptor (sOB-R), and visfatin were measured by ELISA. Results: The mRNA expression and protein content of AdipoR1 and APPL1 did not differ significantly with obesity, age, or puberty. However, GTPase Rab5 protein was increased in the adipocytes of younger prepubertal children with obesity but decreased in AO. Leptin was increased in AO compared to lean adolescents (AL) and in older prepubertal lean (OPL) children and AL compared to younger prepubertal lean and obese children. sOB-R was higher in OPL children and in the AL and AO. Serum visfatin was increased in the younger prepubertal children and AO. Conclusions: In contrast to adults, obesity did not change the expression of AdipoR1 and APPL1 in cultured adipocytes from biopsies of subcutaneous abdominal adipose tissue of children and adolescents. Similar to adipose tissue studies in adults with obesity and metabolic dysfunction, the AO in our study showed reduced adipocyte GTPase Rab5 expression.

Adiponectin and its receptor genes' expression in response to Marek's disease virus infection of White Leghorns

Marek’s disease virus (MDV) causes T-cell lymphoma in susceptible chicken and is also related to an imbalance of the lipid metabolism. Adiponectin is a circulatory cytokine secreted from adipose tissue and exerts critical metabolic functions. Although the associations between adiponectin and diseases, including lipid disorder and noncardiac vascular diseases, have been reported, little is known about the relationship between MDV infection and adiponectin. Here, we challenged white Leghorns from Marek’s disease (MD)-susceptible and MD-resistant lines with MDV at 7 D of age and then explored the body weight and plasma lipoprotein levels at 21 D after MDV infection. Meanwhile, adiponectin and the expression of its receptors were detected using quantitative real-time PCR and Western blot. The results showed that MDV infection induced body weight loss in all the experimental birds. Meanwhile, the concentrations of total cholesterol and high-density lipoprotein were lower after the infection, although there was no significant difference (P > 0.05). However, the infection did not affect adiponectin circulating levels in plasma. MD-susceptible birds had much lower plasma adiponectin than MD-resistant birds (P < 0.01). In abdominal fat, there was no significant difference in adiponectin mRNA level. Still, we observed a significant decrease in adiponectin protein concentration, as well as adipoR1 and adipoR2, at both mRNA and protein levels in the infected compared with the noninfected MD-susceptible chickens. In the spleen, MDV infection significantly reduced the adiponectin mRNA expression but increased the protein in MD-susceptible chickens, which decreased both adipoR1 mRNA expression and protein levels. Also interestingly, the adipoR1 mRNA expression level was significantly increased in MD-susceptible chickens in the liver after MDV infection. All findings in the present study provided interesting insights into adiponectin metabolism in chickens after MDV infection, which helps to advance the understanding of lipid metabolism in response to herpesvirus infection.

Effects of glucose, insulin, glucagon, LPS and poly (I:C) on adiponectin receptors expression in grass carp (Ctenophcuyngodon idellus)

Adiponectin plays critical roles in energy metabolism and immunity of mammals by binding to its receptors (AdipoR1 and AdipoR2). However, they have not been fully understood in fish. The present study examined the distribution of adiponectin receptors (AdipoRs) mRNA in grass carp, and the changes of their expressions under glucose load, insulin or glucagon stimulation, lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid [poly (I:C)] challenge. As for glucose load, fish were orally administrated with glucose (1.67 mg/g BW) for 1, 3, 6, 12 h. For insulin or glucagon stimulation, the primary hepatocytes of grass carp were incubated with 0, 10, 100, 1000 nM insulin or glucagon for 1, 3, 6, 12 h. In the immune challenge experiment, grass carp were intraperitoneally injected with LPS or poly (I:C) (5 μg/g BW) for 3, 6, 12, 24, 48 h. Finally, AdipoRs mRNA expression in all experiments was detected by Real-time quantitative PCR. The results showed that AdipoRs mRNA was widely expressed in various tissues of grass carp. The adipoR1a mRNA was highly expressed in the brain regions, and the highest level of adipoR2 mRNA was observed in the adipose tissue. Glucose load up-regulated adipoR1b and adipoR2 mRNA expressions in the liver at 6 h post administration and decreased adipoR1a expression in the muscle at 3 h. Insulin significantly enhanced adipoR1b and adipoR2 mRNA expressions in the hepatocytes at 1–12 h, and promoted adipoR1a mRNA expression at 1, 3 and 12 h, while glucagon had little effect on their expressions. LPS and poly (I:C) stimuli induced AdipoRs expression in the liver at 6 and 12 h, and their expressions in the muscle were decreased slightly or remained unchanged. In addition, LPS promoted AdipoRs expression in the kidney and spleen, whereas poly (I:C) only induced adipoR1a mRNA expression in the kidney, and increased adipoR2 mRNA level in the spleen. These results suggest that AdipoRs may act as one of the targets of insulin and immune response in grass carp, but whether they are direct target remains to be further studied.

Effects of the adiponectin mimetic compound ALY688 on glucose and fat metabolism in visceral and subcutaneous rat adipocytes

ABSTRACT Adiponectin regulates white adipose tissue (WAT) metabolism and promotes insulin-sensitizing and anti-atherosclerotic effects in vivo. In this context, small molecule adiponectin receptor agonists have become of great therapeutic value for the treatment of metabolic diseases. Here, we investigated the effects of the adiponectin mimetic compound ALY688 on WAT metabolism. To accomplish this, rat epididymal (Epid) and subcutaneous inguinal (Sc Ing) adipocytes were isolated and incubated with ALY688. Subsequently, several parameters of glucose and fat metabolism were assessed. ALY688 promoted AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) phosphorylation, enhanced glucose oxidation, and suppressed fat oxidation in adipocytes from both fat depots. ALY688 did not affect basal and insulin-stimulated rates of glucose uptake, glucose incorporation into lipids, and AKTSer473 and p38 mitogen-activated protein kinase (MAPK) phosphorylations in either Epid or Sc Ing adipocytes. ALY688 did not alter basal lipolysis in Epid and Sc Ing adipocytes, but it enhanced isoproterenol-induced lipolysis in Epid adipocytes. Adiponectin receptor 2 (AdipoR2) mRNA was the prevalent isoform expressed in all adipocytes, and Epid adipocytes displayed significantly higher AdipoR2 mRNA expression than Sc Ing adipocytes. In conclusion, ALY688 can regulate adiposity and affect glycaemic control by altering substrate portioning in the WAT in a fat depot-specific manner.

Adiponectin links maternal metabolism to uterine contractility.

Adiponectin is secreted by adipose tissue and promotes insulin sensitivity. Low circulating adiponectin is associated with increased risk for preterm labor, but the influence of adiponectin on uterine myometrial physiology is unknown. We hypothesized that adiponectin receptors (AdipoRs) decrease myometrial contractility via AMPK to promote uterine quiescence in pregnancy. Using quantitative RT-PCR, we found that nonpregnant or pregnant human and mouse myometrium express AdipoR1 and AdipoR2 mRNAs. We confirmed AdipoR2 protein expression in human and mouse myometrium, with increased abundance in late mouse pregnancy. Both recombinant adiponectin and a pharmacologic AdipoR agonist, AdipoRon, potently inhibited uterine myometrial strip contractions in physiologic organ bath. The relaxation was independent of contractile stimulus (oxytocin, KCl, U46619). AdipoR agonists increased AMPK phosphorylation in pregnant mouse myometrium, and the direct AMPK activator A769662 also relaxed myometrial strips. However, the AMPK inhibitor dorsomorphin (compound C) blocked AMPK phosphorylation but did not abolish relaxation with either AdipoRon or A769662. In summary, adiponectin inhibits myometrial contractility consistent with the possibility that it is a previously unrecognized link between maternal metabolism and pregnancy maintenance. We also identify a separate role for AMPK regulating myometrial contractions that may influence labor onset.-Vyas, V., Guerra, D. D., Bok, R., Powell, T., Jansson, T., Hurt, K. J. Adiponectin links maternal metabolism to uterine contractility.

Intensive Statin Therapy Compromises the Adiponectin-AdipoR Pathway in the Human Monocyte-Macrophage Lineage.

Background and Purpose- Statins are widely used for cardiovascular disease prevention through cholesterol-lowering and anti-inflammatory effects. Adiponectin, an anti-inflammatory adipokine, acts via two receptors, AdipoR1 and AdipoR2, to exert atheroprotective effects on the vasculature. We investigated whether statins can modulate the adiponectin-AdipoR pathway in the human monocyte-macrophage lineage. Methods- Monocytes were isolated from the whole blood of patients with severe carotid atherosclerosis (cross-sectional study) or from patients with cardiovascular risk factors (longitudinal study) and assessed for AdipoR1 and AdipoR2 gene expression using quantitative real-time polymerase chain reaction. In vitro, THP-1 (Tamm-Horsfall protein 1) macrophages were treated with increasing atorvastatin or rosuvastatin doses for 24- or 72-hours to determine the effect of statins on AdipoR expression and activity. Macrophage cytokine secretion (IL [interleukin]-1β, IL-10, IL-6, and TNF [tumor necrosis factor]-α) was assessed by electrochemiluminescence. Results- AdipoR1 and AdipoR2 mRNA expression on circulating monocytes from patients with carotid atherosclerosis, was significantly lower by 1.36- and 1.17-fold, respectively, in statin users versus statin-naïve patients. Specifically, patients on high doses of atorvastatin (40-80 mg) or rosuvastatin (20-40 mg) had significantly lower AdipoR gene expression versus statin-naïve patients. Similarly, in the longitudinal in vivo study, longer atorvastatin/rosuvastatin treatment (≥5 months) in patients with cardiovascular risk factors resulted in lower AdipoR gene expression on circulating monocytes versus prestatin levels. In vitro, higher statin doses and longer exposure resulted in a greater decrease in AdipoR mRNA expression and greater macrophage secretion of pro-inflammatory cytokines, IL-1β, IL-6, and TNF-α. High statin doses also reduced adiponectin's capacity to suppress intracellular cholesteryl ester levels in oxLDL (oxidized LDL)-loaded macrophages, with rosuvastatin exhibiting higher potency than atorvastatin. Conclusions- Our in vivo and in vitro studies identified a novel pleiotropic property of statins in modulating the adiponectin-AdipoR pathway in the human monocyte-macrophage lineage, where intensive statin therapy compromised the expression and function of adiponectin and its receptors.

Moderate consumption of fermented alcoholic beverages diminishes diet-induced non-alcoholic fatty liver disease through mechanisms involving hepatic adiponectin signaling in mice

PurposeResults of some epidemiological studies suggest that moderate alcohol consumption may be associated with a decreased risk to develop NAFLD. Here, the effect of the consumption of moderate beer and diluted ethanol, respectively, on the development of NAFLD were assessed.MethodsFemale C57BL/6J mice were fed a control diet (C-D) or a diet rich in fructose, fat and cholesterol (FFC) enriched isocalorically and isoalcoholically with beer (FFC + B) or plain ethanol (FFC + E) (2.5 g ethanol/kg body weight/day) for 7 weeks. Liver damage was assessed by histology using NAFLD activity score. Markers of inflammation, insulin resistance and adiponectin signaling were measured at mRNA and protein levels. Using J774A.1 cells as a model of Kupffer cells, the effect of alcoholic beverages on adiponectin receptor 1 (Adipor1) was assessed.ResultsHepatic triglyceride concentration, neutrophil granulocytes, iNOS protein concentrations and early signs of insulin resistance found in FFC-fed mice were significantly attenuated in FFC+ B-fed mice (P < 0.05 for all). These findings were associated with a super-induction of Adipor1 mRNA expression (+ ~ 18-fold compared to all other groups) and a decrease of markers of lipid peroxidation in liver tissue of FFC + B-fed mice when compared to FFC-fed animals. Similar differences were not found between FFC– and FFC+ E-fed mice. Expression of Adipor1 was also super-induced (7.5-fold) in J774A.1 cells treated with beer (equivalent to 2 mmol/L ethanol).ConclusionsThese data suggest that moderate intake of fermented alcoholic beverages such as beer at least partially attenuates NAFLD development through mechanisms associated with hepatic AdipoR1 expression.

Comparing pig breeds with genetically low and high backfat thickness: differences in expression of adiponectin, its receptor, and blood metabolites

Here we characterized gene expressions in subcutaneous adipose tissue and blood metabolites of pigs with genetically low backfat (Landrace) and high backfat (Meishan). As pigs aged from 1 wk-to 3-mo old, mRNA levels of adipose-specific genes increased, although their gene expressions coding for major enzymes involved in lipid metabolism (lipoprotein lipase, fatty acid synthase, and hormone-sensitive lipase) did not differ between lean and fat pigs. Instead, there were significant effects for adiponectin and its receptor AdipoR1 mRNA levels between the two breeds of which respective expressions were lower and higher in Meishan by 3 mo of age. Contrary to changes in gene expressions, the concentrations of blood glucose, triglyceride (TG), and NEFA in both breeds decreased during growth, and 3-mo-old Meishan evidenced lower glucose with higher TG than the Landrace. The homeostasis model assessment insulin resistance (HOMA-IR) index was also calculated from the measurements of fasting glucose and insulin concentration, and Meishan showed a higher value than the Landrace. We next examined these differences in Landrace and Meishan crossbreds, which were phenotypically distinguishable by the backfat thickness as the former lean type and the latter fat type. As with the purebreds, high backfat Meishan crosses showed the characteristics of lower glucose and higher TG in circulating levels and also lower adiponectin transcripts in subcutaneous adipose tissue. Collectively, our results demonstrate that levels of adiponectin and its receptor gene expressions, blood glucose, blood lipids, and HOMA-IR in pigs vary between lean and fat. These observations strongly suggest the possibility that overall metabolic differences rather than adipocyte ability itself contribute to the fatness of genetically high backfat pigs.

Recombinant adiponectin alleviates abortion in mice by regulating Th17/Treg imbalance via p38MAPK-STAT5 pathway

Recurrent spontaneous abortion is associated with abnormal maternal tolerance to the semi-allogenic fetus, wherein the Th17/Treg axis plays a crucial role. Adiponectin (APN) is an adipocytokine that is shown to be a novel negative T-cell regulator and induce immune tolerance. The CBA/J×DBA/2 mating was used as an abortion-prone model to investigate whether the addition of recombinant adiponectin (rAPN) improves the pregnancy outcome. Recombinant adiponectin therapy reduced the abortion rate in abortion-prone model. It skewed the ability of serum cytokine production toward a Treg bias and induced APN production. Flow cytometry revealed that rAPN administration expanded the splenic CD4+CD25+ regulatory T-cell (Treg) population and reduced the Th17 cell populations in CBA/J×DBA/2 matings. RT-PCR revealed that rAPN administration induced the expression of AdipoR1 and AdipoR2 mRNA at the maternofetal interface. Recombinant adiponectin administration induced FoxP3 and reduced RORγt expressions at the maternofetal interface. In vitro experiment also showed that rAPN treatment enhanced the FoxP3 mRNA and protein expression and decreased the RORγt expression in splenic lymphocytes of abortion-prone mice. Blocking the different signal transduction pathways downstream of APN, p38MAPK inhibitor (SB203580) and STAT5 inhibitor (Pimozide) could abrogate the regulatory effect of rAPN on FoxP3 and RORγt expression, while STAT3 inhibitor (Stattic) and AMPK inhibitor (p5499) did not exert any influence. Thus, the current results demonstrated that rAPN therapy improves pregnancy outcome in a murine model of abortion by expanding the Treg cell population and function and decreasing the Th17 cell population and function via a p38MAPK-STAT5 pathway.

Mango leaf tea promotes hepatoprotective effects in obese rats

As mango leaf tea contains mangiferin and other bioactive compounds, this study investigated its anti-inflammatory, antioxidant and hepatoprotective effects on rats with high-fat induced obesity. Three groups were established: a control group (AIN93M diet), an obese group (high-fat diet HFD) and a treatment group (HFD with mango leaf tea for 8 weeks). Mango leaf tea increased antioxidant enzymes, total antioxidant capacity, AdipoR2 and PPAR-α mRNA and proteins expressions and, it also inhibited the NF-κB p65 and SREBP1c genes expressions in the liver. This beverage also leads to Cpt1 overexpression and a significant decrease in the accumulation of fat droplets, improving the hepatic steatosis. Molecular docking suggested a positive interaction between mangiferin, the main bioactive compound of mango leaf tea, and PPAR-α. Mango leaf tea exhibited a hepatoprotective effect through activating PPARα and decreasing the NF-κB p65 expressions, reducing oxidative stress and steatosis, and improving the lipid metabolism.

Effects of maternal obesity on maternal and fetal plasma concentrations of adiponectin and expression of adiponectin and its receptor genes in cotyledonary and adipose tissues at mid- and late-gestation in sheep

Adiponectin potentially influences fetal weight by altering insulin signaling and trans-placental amino acid and glucose transporters. The objective of this study was to determine how maternal obesity influences maternal and fetal plasma concentrations of adiponectin, expression of fetal adiponectin, its receptors, and adipogenic genes at mid- and late-gestation. Blood samples and tissues were collected from obese and control multiparous pregnant ewes at day 75 or 135 of gestation. Although day of gestation or maternal obesity did not influence (P > 0.6) maternal plasma concentrations of adiponectin, fetal weight was increased (P < 0.001) and adiponectin tended to decrease (P = 0.10) at mid-gestation in fetuses from obese ewes. Differences were not apparent at late-gestation (P > 0.70). Relative abundance of adiponectin (P = 0.01), AdipoR2 (P = 0.04) and PPARγ (P = 0.01) mRNA was less at mid-gestation in fetal adipose tissue from obese mothers. By late gestation, maternal obesity tended to associated with a decrease in relative abundance of adiponectin (P = 0.09) and SREBF1 (P = 0.10) mRNA in fetal adipose tissue. Maternal obesity did not influence (P ≥ 0.20) the relative abundance of adiponectin, AdipoR1 and AdipoR2 mRNA in cotyledonary tissue at mid or late- gestation. In conclusion, maternal obesity in sheep influences relative abundance of fetal adipose tissue mRNA for adiponectin and adipogenic, as well as plasma concentrations of total adiponectin. Although adiposity in pregnant ewes did not influence maternal adiponectin, maternal obesity potentially influenced fetal adipogenesis by altering the abundance of adiponectin, PPARγ and SREBF1 mRNA in fetal adipose tissue.

Effect of AMPK/mTOR/S6K1 pathways and the insulin-sensitizing effect for adiponectin in endometrial cancer cells

Objective: To explore effect of AMP-activated protein kinase (AMPK) /mammalian target of rapamycin (mTOR) /ribosomal protein S6 kinase-1 (S6K1) signaling pathways and the insulin-sensitizing effect by adiponectin in endometrial cancer HEC-1B cells. Methods: The experiments were divided into 4 groups, adiponectin (Ad) group (HEC-1B cells treated with 20 μg/ml adiponectin for 30 minutes) , inhibitor group (HEC-1B cells treated with 10 μmol/L compound C for 30 minutes) , inhibitor+ Ad group (HEC-1B cells treated with 10 μmol/L compound C for 30 minutes following incubation with 20 μg/ml adiponectin for 30 minutes) , control group (only added the culture medium without serum DMEM) . (1) Real-time quantitative PCR and western blot analysis were used to detect the level of mRNA and protein of adiponectin receptor (AdipoR) 1 and AdipoR2. (2) Western blot analysis were used to detect phosphorylation of AMPK, mTOR, S6K1 or insulin receptor substrate 1 (IRS1) protein expression with stimulation in different concentrations of adiponectin (2.5, 5, 10, and 20 μg/ml) , or following incubation with insulin 50 nmol/L for 5 minutes; or treated with 20 μg/ml adiponectin for different times (15, 30, 45, and 60 minutes) , or following incubation with insulin 50 nmol/L for 5 minutes. (3) Cell counting kit-8 (CCK-8) assay was performed to investigate the cell proliferation, and transwell chamber assay was used to detect the cell migration in different groups. Results: (1) The relative expression level of AdipoR1 mRNA and protein were higher than AdipoR2 in HEC-1B cell (8.50±0.09 to 1.00±0.00, and 0.91±0.03 to 0.69±0.03; P<0.05) . (2) The phosphorylation level of p-AMPK was significantly induced, and the phosphorylation level of p-mTOR and p-S6K1 proteins, and 20 μg/ml adiponectin at 30 minutes, AMPK protein had the highest level of activation. (3) Adiponectin induces increased tyrosine phosphorylation of IRS1 in a time-and concentration-dependent manner. (4) The proliferation inhibition ratio in Ad group (0.68±0.34) % was much more than that in inhibitor+Ad group (0.24±0.04) % (t=17.88, P<0.05) . The number of cell migration in Ad group (77±8) was much more than that in inhibitor+Ad group (132±13; t=-7.34, P<0.05) . Conclusions: Adiponectin maybe inhibit proliferation and migration of endometrial cancer cells through AMPK/mTOR/S6K1 signal pathway. Adiponectin insensitizes insulin signaling may by regulating by the AMPK/S6K1/IRS1 pathway.

MiR‑6835‑3p regulates the function of pancreatic islet cells by modulating the expression of AdipoR1.

Effective drugs and strategies for treating type 2 diabetes mellitus (2-DM) are urgently required. The aim of the present study was to elucidate the mechanism underlying microRNA (miR)-6835-3p regulation of adiponectin receptor 1 (AdipoR1) expression and the miR-6835-3p/AdipoR1 signaling pathway in pancreatic islet cells. In addition, the potential anti-diabetes effect of miR-6835-3p on insulin secretion was investigated. Luciferase activity analysis was performed to evaluate how miR-6835-3p targets the 3′-untranslated region of AdipoR1. The SU.86.86 and MIN-6 cell lines were co-cultured with or without miR-6835-3p inhibitors or mimics, and the insulin secretory functions of these cell lines were then determined. Luciferase reporter analysis revealed that AdipoR1 was a direct target of miR-6835-3p. In addition, miR-6835-3p overexpression suppressed the mRNA and protein expression levels of AdipoR1 in the SU.86.86 and MIN-6 cell lines. Furthermore, miR-6835-3p exerted negative effects on insulin secretion in SU.86.86 and MIN-6 cells, which were mediated by regulating AdipoR1 expression. AdipoR1 was a direct target of miR-6835-3p; therefore, inhibition of AdiopR1 expression may reduce insulin secretion and may be considered a key regulator of insulin secretion. The results of the present study suggested that targeting AdipoR1 with miR-6835-3p inhibitors may be a potential strategy for promoting glucose-stimulated insulin secretion, and thereby, may be an effective treatment for type 2-DM.

Effects of vitamin D-induced chronic renal disease on quality and cryopreservation of spermatozoa from the endangered Iberian lynx

Adiponectin potentially influences fetal weight by altering insulin signaling and trans-placental amino acid and glucose transporters. The objective of this study was to determine how maternal obesity influences maternal and fetal plasma concentrations of adiponectin, expression of fetal adiponectin, its receptors, and adipogenic genes at mid- and late-gestation. Blood samples and tissues were collected from obese and control multiparous pregnant ewes at day 75 or 135 of gestation. Although day of gestation or maternal obesity did not influence (P > 0.6) maternal plasma concentrations of adiponectin, fetal weight was increased (P < 0.001) and adiponectin tended to decrease (P = 0.10) at mid-gestation in fetuses from obese ewes. Differences were not apparent at late-gestation (P > 0.70). Relative abundance of adiponectin (P = 0.01), AdipoR2 (P = 0.04) and PPARγ (P = 0.01) mRNA was less at mid-gestation in fetal adipose tissue from obese mothers. By late gestation, maternal obesity tended to associated with a decrease in relative abundance of adiponectin (P = 0.09) and SREBF1 (P = 0.10) mRNA in fetal adipose tissue. Maternal obesity did not influence (P ≥ 0.20) the relative abundance of adiponectin, AdipoR1 and AdipoR2 mRNA in cotyledonary tissue at mid or late- gestation. In conclusion, maternal obesity in sheep influences relative abundance of fetal adipose tissue mRNA for adiponectin and adipogenic, as well as plasma concentrations of total adiponectin. Although adiposity in pregnant ewes did not influence maternal adiponectin, maternal obesity potentially influenced fetal adipogenesis by altering the abundance of adiponectin, PPARγ and SREBF1 mRNA in fetal adipose tissue.

Upregulation of the adipokine genes ADIPOR1 and SPP1 is related to poor survival outcomes in colorectal cancer.

Obesity is closely associated with colorectal cancer (CRC), but the underlying mechanism is unclear. We thus evaluated the expression of the adipokine gene family in CRC tissues and its clinicopathological implications. Correlations between the mRNA expression levels of the adipokine gene family (ADIPOQ, ADIPOR1/2, LEP, LEPR, RETN, RETNLB, RBP4, SFRP5, NAMPT, and SPP1) in CRC tissue and clinicopathologic factors were analyzed using data from The Cancer Genome Atlas database. Tissue samples from 369 patients were analyzed, and 82 deaths occurred during follow-up (median, 670 days). Overall, mortality was associated with positive venous invasion, higher TNM stage, and increased ADIPOR1 (adiponectin receptor 1 gene) and SPP1 (secreted phosphoprotein gene 1) mRNA expression. Higher ADIPOR1 (odds ratio [OR]: 3.29, 95% confidence interval [CI]: 1.33-8.13) and SPP1 (OR: 2.31, 95%CI: 1.49-3.59) levels were independently associated with increased mortality. A Kaplan-Meier survival analysis showed shorter overall survival times in patients with higher ADIPOR1 (P = 0.006) and SPP1 (P < 0.001) expression. Upregulation of ADIPOR1 and SPP1, among the adipokine gene family, in cancer tissue is associated with poor survival in CRC, suggesting a potential mechanism linking obesity and CRC. ADIPOR1 and SPP1 expression could become useful prognostic indicators after further validation.

Abstract 125: Intensive Statin Therapy Negatively Modulates AdipoR Activity in the Monocyte-macrophage Lineage in vivo and in vitro

Introduction: Statins are widely used for primary and secondary prevention of cardiovascular disease and its complications through cholesterol lowering and anti-inflammatory effects. Adiponectin, an anti-inflammatory adipokine, acts via 2 receptors, AdipoR1 and AdipoR2, to exert protective effects on the vasculature. Recently, we demonstrated that AdipoR are highly expressed on macrophages and that decreased AdipoR2 activity is associated with atherosclerotic plaque instability. We aimed to investigate whether statins can modulate AdipoR activity in the monocyte-macrophage lineage. Methods: Blood monocytes were isolated from whole blood of patients (statin-naïve vs statin users) with severe carotid atherosclerosis by a Magnetic Cell-Sorting technique. AdipoR1 and AdipoR2 mRNA expression on circulating monocytes was determined using qRT-PCR. In vitro , THP-1 macrophages were treated for 24 or 72 hours with varying doses of atorvastatin or rosuvastatin (1μM, 10μM, 60μM) to determine the effect of statins on AdipoR activity (AMPK, PPAR-α) by qRT-PCR and Western Blot analyses. Macrophage cytokine secretion (IL-1β, IL-10, IL-6, TNF-α) was assessed by electrochemiluminescence. Results: AdipoR1 and AdipoR2 mRNA expression on circulating monocytes was significantly lower by 1.36- and 1.17-fold (P&lt;0.05), respectively, in statin users (n=157) compared to statin-naïve patients (n=37). When analyzing by statin type and dose, only patients on high doses of atorvastatin (40-80 mg) or rosuvastatin (20-40 mg) had significantly lower AdipoR expression compared to statin-naïve patients (P&lt;0.05). Statins effects on lowering AdipoR expression was only apparent in patients with unstable but not stable plaques. In vitro , higher doses and longer exposure of macrophages to atorvastatin or rosuvastatin resulted in a greater decrease in AdipoR mRNA expression and signalling through AMPK and PPAR-α, and greater secretion of pro-inflammatory cytokines, IL-1β, IL-6, and TNF-α. Conclusion: In our study, intensive statin therapy resulted in negative modulation of AdipoR expression and function. Despite their known protective properties, statins may also elicit severe adverse effects, which are often associated with long-term treatment and high doses.

Chronic Intermittent Hypoxia in Premature Infants: The Link Between Low Fat Stores, Adiponectin Receptor Signaling and Lung Injury.

Premature infants have chronic intermittent hypoxia (CIH) that increases morbidity, and the youngest and the smallest premature infants are at the greatest risk. The combination of lung injury from inflammation/oxidative stress causing low functional residual capacity combined with frequent short apneas leads to CIH. Adiponectin (APN) is an adipose-derived adipokine that protects the lung from inflammation and oxidative stress. Premature and small for gestational age (SGA) infants have minimal body fat and low levels of circulating APN. To begin to understand the potential role of APN in lung protection during lung development, we characterized the developmental profile of APN and APN receptors (AdipoR1 and AdipoR2) protein and mRNA expression in the newborn rat lung at fetal day (FD) 19, and postnatal days (PD) 1, 4, 7, 10, 14, 21, and 28. Protein levels in lung homogenates were measured by western blot analyses; relative mRNA expression was detected by quantitative PCR (qPCR); and serum high molecular weight (HMW) APN was measured using enzyme-linked immunosorbent assay (ELISA). Results: APN protein and mRNA levels were lowest at FD19 and PD1, increased 2.2-fold at PD4, decreased at PD10, and then increased again at PD21. AdipoR1 protein and mRNA levels peaked at PD1, followed by a threefold drop by PD4, and remained low until PD21. AdipoR2 protein and mRNA levels also peaked at PD1, but remained high at PD4, followed by a 1.7-fold drop by PD10 that remained low by PD21. Serum APN levels detected by ELISA did not differ from PD4 to PD28. To date, this is the first report characterizing APN and APN receptor protein and mRNA expression in the rat lung during development. The developmental stage of the newborn rat lung models that of the premature human infant; both are in the saccular stage of lung development. In the newborn rat lung, alveolarization begins at PD4, peaks at PD10, and ends at PD21. Importantly, we found that AdipoR1 receptor protein and mRNA expression is lowest during lung alveolarization (PD4 to PD21). Thus, we speculate that low levels of AdipoR1 during lung alveolarization contributes to the increased susceptibility to developing acute lung edema and chronic lung injury such as bronchopulmonary dysplasia (BPD) in premature human infants.

Cannabidiol attenuates alcohol-induced liver steatosis, metabolic dysregulation, inflammation and neutrophil-mediated injury

Cannabidiol (CBD) is a non-psychoactive component of marijuana, which has anti-inflammatory effects. It has also been approved by FDA for various orphan diseases for exploratory trials. Herein, we investigated the effects of CBD on liver injury induced by chronic plus binge alcohol feeding in mice. CBD or vehicle was administered daily throughout the alcohol feeding study. At the conclusion of the feeding protocol, serums samples, livers or isolated neutrophils were utilized for molecular biology, biochemistry and pathology analysis. CBD significantly attenuated the alcohol feeding-induced serum transaminase elevations, hepatic inflammation (mRNA expressions of TNFα, MCP1, IL1β, MIP2 and E-Selectin, and neutrophil accumulation), oxidative/nitrative stress (lipid peroxidation, 3-nitrotyrosine formation, and expression of reactive oxygen species generating enzyme NOX2). CBD treatment also attenuated the respiratory burst of neutrophils isolated from chronic plus binge alcohol fed mice or from human blood, and decreased the alcohol-induced increased liver triglyceride and fat droplet accumulation. Furthermore, CBD improved alcohol-induced hepatic metabolic dysregulation and steatosis by restoring changes in hepatic mRNA or protein expression of ACC-1, FASN, PPARα, MCAD, ADIPOR-1, and mCPT-1. Thus, CBD may have therapeutic potential in the treatment of alcoholic liver diseases associated with inflammation, oxidative stress and steatosis, which deserves exploration in human trials.

Effects of (-)-hydroxycitric acid on lipid droplet accumulation in chicken embryos.

This study was conducted to determine the impact of (-)-hydroxycitric acid ((-)-HCA) on biochemical indices and lipid metabolism parameters in chicken embryos. Two hundred and forty fertilized eggs were divided into six groups and injected with (-)-HCA at concentrations of 0, 0.1, 0.5, 1.0, 10.0 and 50mg/kg (n=40). After 19days of incubation, serum and liver were collected for analysis of biochemical indices and lipid metabolism parameters. Results showed no significant differences on serum biochemical indices: 1-50mg/kg (-)-HCA significantly increased serum glucose and hepatic glycogen contents (P<0.05). Oil Red O staining analysis showed total area, counts of lipid droplets and hepatic triglyceride content were significantly decreased (P<0.01), meanwhile hepatic lipase and lipoprotein lipase activity were significantly increased (P<0.05). ACLY, ME1, SREBP-1c messenger RNA (mRNA) levels in 0.5-10mg/kg groups and FAS mRNA level in 1-10mg/kg groups were significantly decreased (P<0.05), while PPARα mRNA level, serum adiponectin content and AdipoR1 mRNA level were significantly increased in 0.5-50mg/kg groups (P<0.05). These results indicated (-)-HCA treatment inhibited triglyceride synthesis via decreasing lipogenesis-related factors, mRNA expression level and accelerated lipolysis by enhancing lipoprotein lipase and hepatic lipase activity, which finally reduced lipid droplet accumulation, and this action may be associated with activating the adiponectin signaling pathway.