Liver Alpha Research Articles

Estrogen receptor alpha in the female liver: Dr. Jekyll and Mr. Hyde

Estrogen receptor alpha in the female liver: Dr. Jekyll and Mr. Hyde

Pattern of Alpha Fetoprotein and Activities of Liver Enzymes in Type 2 Diabetic Patients Attending Nnamdi Azikiwe University Teaching Hospital Nnewi, Anambra State, Nigeria.

Diabetes mellitus (DM) is a metabolic disorder characterized by hyperglycemia, which has become health problem of global interest, linked to renal disease. The study is to assess the activities of some liver enzymes and alpha fetoprotein level in type 2 diabetic patients. 81 participants comprised 38 diabetics and 43 controls were randomly recruited. Informed, oral and written consent was obtained from the participants. Ethical approval was obtained. 6mls of fasting blood samples were collected. Plasma glucose, alpha feto-protein (AFP) levels; serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and gamma glutamyltransferase (GGT) activities were analyzed using standard laboratory methods. The results showed significantly increased plasma glucose level in the diabetics than in controls (p=0.000), as well as in male diabetics than in female diabetics (p=0.048). But, the mean age, AFP, ALT, AST, ALP and GGT activities were the same in the diabetics and the controls; and in male and female diabetics at p>0.05 in each case. Relatively strong positive significant correlation was observed between ALT Vs AST (r=0.566; p=0.000) and a weaker negative association exist between ALT Vs Age (r= -0.353; p=0.022) in diabetics but no associations were observed with other parameters studied. In conclusion the positive association between ALT and AST may reveal transaminase derangements in liver status of type 2 diabetic subjects studied.

Effects of genetic selection on activity of corticotropic and thyrotropic axes in modern broiler chickens

Commercial selection for meat-type (broiler) chickens has produced economically valuable birds with fast growth rates, enhanced muscle mass, and highly efficient feed utilization. The physiological changes that account for this improvement and unintended consequences associated with them remain largely unexplored, despite their potential to guide further advancements in broiler production efficiency. To identify effects of genetic selection on hormonal signaling in the adrenocorticotropic and thyrotropic axes, gene expression in muscle and liver and post-hatch circulating hormone concentrations were measured in legacy [Athens Canadian Random Bred (ACRB)] and modern (Ross 308) male broilers between embryonic days (e) 10 and e18 and post-hatch days (d) 10 and d40. No interactive effects or main effects of line were observed for adrenocorticotropic gene expression during either developmental period, although age effects appeared for corticosteroid-binding globulin in liver during embryogenesis and post-hatch and glucocorticoid receptor in both tissues post-hatch. There was a main line effect for circulating corticosterone, with levels in ACRB greater than those in Ross. Several thyrotropic genes exhibited line-by-age interactions during embryonic or post-hatch development. In liver, embryonic expression of thyroid hormone receptor beta was greater in ACRB on e12, and deiodinase 3 (DIO3) levels were greater in Ross on e14 and e16. In juvenile liver, deiodinase 2 (DIO2) expression was greater in ACRB on d10 but greater in Ross on d20, while DIO3 was higher in ACRB on d30 and d40. Levels of thyroid hormone receptor alpha mRNA exhibited a main line effect, with levels greater in ACRB juvenile breast muscle. Several thyrotropic genes exhibited main age effects, including DIO2 and DIO3 in embryonic breast muscle, thyroid hormone receptor alpha and thyroid hormone receptor beta in post-hatch liver, and DIO2 in post-hatch breast muscle. Circulating triiodothyronine displayed a main line effect, with levels in Ross significantly reduced as compared to ACRB. These findings suggest that in modern broilers, a decrease in levels of hormones that control basal metabolism triiodothyronine and the stress response circulating corticosterone, as well as altered expression of genes regulating thyroid hormone activity, could contribute to lower heat production, reduced stress response, and altered nutrient partitioning, leading to more efficient feed utilization and faster, more productive growth.

Co-expression of drug metabolizing cytochrome P450 enzymes and estrogen receptor alpha (ESR1) in human liver: racial differences and the regulatory role of ESR1.

The function and expression of cytochrome P450 (CYP) drug metabolizing enzymes is highly variable, greatly affecting drug exposure, and therapeutic outcomes. The expression of these enzymes is known to be controlled by many transcription factors (TFs), including ligand-free estrogen receptor alpha (ESR1, in the absence of estrogen). However, the relationship between the expression of ESR1, other TFs, and CYP enzymes in human liver is still unclear. Using real-time PCR, we quantified the mRNA levels of 12 CYP enzymes and nine TFs in 246 human liver samples from European American (EA, n = 133) and African American (AA, n = 113) donors. Our results showed higher expression levels of ESR1 and six CYP enzymes in EA than in AA. Partial least square regression analysis showed that ESR1 is the top-ranking TF associating with the expression of eight CYP enzymes, six of which showed racial difference in expression. Conversely, four CYP enzymes without racial difference in expression did not have ESR1 as a top-ranking TF. These results indicate that ESR1 may contribute to variation in CYP enzyme expression between these two ancestral backgrounds. These results are consistent with our previous study showing ESR1 as a master regulator for the expression of several CYP enzymes. Therefore, factors affecting ESR1 expression may have broad influence on drug metabolism through altered expression of CYP enzymes.

A Pre-Operative Prognostic Score for Patients With Advanced Hepatocellular Carcinoma Who Underwent Resection.

BackgroundPrevious studies demonstrated a promising prognosis in advanced hepatocellular carcinoma (HCC) patients who underwent surgery, yet a consensus of which population would benefit most from surgery is still unreached.MethodA total of 496 advanced HCC patients who initially underwent liver resection were consecutively collected. Least absolute shrinkage and selection operator (LASSO) regression was performed to select significant pre-operative factors for recurrence-free survival (RFS). A prognostic score constructed from these factors was used to divide patients into different risk groups. Survivals were compared between groups with log-rank test. The area under curves (AUC) of the time-dependent receiver operating characteristics was used to evaluate the predictive accuracy of prognostic score.ResultFor the entire cohort, the median overall survival (OS) was 23.0 months and the median RFS was 12.1 months. Patients were divided into two risk groups according to the prognostic score constructed with ALBI score, tumor size, tumor-invaded liver segments, gamma-glutamyl transpeptidase, alpha fetoprotein, and portal vein tumor thrombus stage. The median RFS of the low-risk group was significantly longer than that of the high-risk group in both the training (10.1 vs 2.9 months, P<0.001) and the validation groups (13.7 vs 4.6 months, P=0.002). The AUCs of the prognostic score in predicting survival were 0.70 to 0.71 in the training group and 0.71 to 0.72 in the validation group.ConclusionSurgery could provide promising survival for HCC patients at an advanced stage. Our developed pre-operative prognostic score is effective in identifying advanced-stage HCC patients with better survival benefit for surgery.

The liver\u2013alpha cell axis associates with liver fat and insulin resistance: a validation study in women with non-steatotic liver fat levels

Aims/hypothesisMany individuals who develop type 2 diabetes also display increased glucagon levels (hyperglucagonaemia), which we have previously found to be associated with the metabolic syndrome. The concept of a liver–alpha cell axis provides a possible link between hyperglucagonaemia and elevated liver fat content, a typical finding in the metabolic syndrome. However, this association has only been studied in individuals with non-alcoholic fatty liver disease. Hence, we searched for a link between the liver and the alpha cells in individuals with non-steatotic levels of liver fat content. We hypothesised that the glucagon–alanine index, an indicator of the functional integrity of the liver–alpha cell axis, would associate with liver fat and insulin resistance in our cohort of women with low levels of liver fat.MethodsWe analysed data from 79 individuals participating in the Prediction, Prevention and Subclassification of Type 2 Diabetes (PPSDiab) study, a prospective observational study of young women at low to high risk for the development of type 2 diabetes. Liver fat content was determined by MRI. Insulin resistance was calculated as HOMA-IR. We conducted Spearman correlation analyses of liver fat content and HOMA-IR with the glucagon–alanine index (the product of fasting plasma levels of glucagon and alanine). The prediction of the glucagon–alanine index by liver fat or HOMA-IR was tested in multivariate linear regression analyses in the whole cohort as well as after stratification for liver fat content ≤0.5% (n = 39) or >0.5% (n = 40).ResultsThe glucagon–alanine index significantly correlated with liver fat and HOMA-IR in the entire cohort (ρ = 0.484, p < 0.001 and ρ = 0.417, p < 0.001, respectively). These associations resulted from significant correlations in participants with a liver fat content >0.5% (liver fat, ρ = 0.550, p < 0.001; HOMA-IR, ρ = 0.429, p = 0.006). In linear regression analyses, the association of the glucagon–alanine index with liver fat remained significant after adjustment for age and HOMA-IR in all participants and in those with liver fat >0.5% (β = 0.246, p = 0.0.23 and β = 0.430, p = 0.007, respectively) but not in participants with liver fat ≤0.5% (β = −0.184, p = 0.286).Conclusions/interpretationWe reproduced the previously reported association of liver fat content and HOMA-IR with the glucagon–alanine index in an independent study cohort of young women with low to high risk for type 2 diabetes. Furthermore, our data indicates an insulin-resistance-independent association of liver fat content with the glucagon–alanine index. In summary, our study supports the concept that even lower levels of liver fat (from 0.5%) are connected to relative hyperglucagonaemia, reflecting an imminent impairment of the liver–alpha cell axis.

Nicorandil and atorvastatin attenuate carbon tetrachloride – induced liver fibrosis in rats

Purpose The present study aimed to evaluate the possible hepatoprotective effects of nicorandil and atorvastatin against experimentally induced liver fibrosis. Materials and methods Wistar male rats wereassigned tofivegroups; control group, fibrosis group, the remaining three groups received in addition to CCl4, N-acetyl cysteine (300 mg/kg), nicorandil(15 mg/kg) and atorvastatin (20 mg/kg), respectively. Liver fibrosis was induced by intraperitoneal injection of rats with CCl4 (2 ml/kg), twice weekly for five consecutive weeks. All treatments were administered daily starting from the first day of fibrosis induction for five consecutive weeks. By the end of the experiment, fibrosis biomarkers [hepatic transforming growth factor β1 (TGF-β1) and hydroxyproline (HYP)], liver function [serum alanine transaminase (ALT), aspartate transaminase (AST), albumin and total bilirubin] were assessed. Moreover, lipid profile [total cholesterol, serum triglycerides, high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C)], inflammatory biomarkers [hepatic myeloperoxidase (MPO), serum tumor necrosis factor alpha (TNF-α)], relative liver weight] and oxidative stress biomarkers [malondialdehyde (MDA), glutathione (GSH) and catalase (CAT)] were evaluated. In support, histopathological and immunohistochemical examination of liver alpha smooth muscle actin (α-SMA) were performed. Results Nicorandil and atorvastatin effectively reduced fibrosis and liver function biomarkers. They both restored serum lipid profile, TNF-α, MPO, relative liver weight, and hepatic MDA content. Alternatively, they markedly elevated albumin, HDL-C and hepatic content of GSH and CAT. Additionally, a marked histopathological and immunohistochemical improvement of α-SMA was observed. Conclusion Nicorandil and atorvastatin might be promising protective agents against liver fibrosis through amelioration of liver function, modulation of fibrous formation, anti-inflammatory and antioxidant potentials.

Fasting Increases Iron Export by Modulating Ferroportin 1 Expression Through the Ghrelin/GHSR1α/MAPK Pathway in the Liver.

The liver is contributed to maintaining body iron homeostasis and controlling of body adaptation to fasting. Although previous studies implied a negative relationship between iron and ghrelin in both mice and humans, it remains to be explored whether fasting or ghrelin has a functional effect on iron homeostasis in the liver. In this study, we examined the roles of fasting and ghrelin in modulating the protein expression of Fpn1, transferrin receptor 1 (TfR1), and ferritin light chain (Ft-L), as well as the mRNA expression of ghrelin, hepcidin, ghrelin O-acyltransferase (GOAT), and growth hormone secretagogue receptor 1 alpha (GHSR1α) in mouse liver and cultured hepatocytes. Our in vivo results suggested that fasting significantly upregulated the mRNA expression of ghrelin, GOAT, and GHSR1α, as well as the protein levels of ghrelin, Fpn1, and Ft-L, but not TfR1, in mouse liver. Interestingly, mRNA expression of hepcidin did not change significantly after fasting. Meanwhile, in cultured hepatocytes, ghrelin significantly increased the protein expression of Fpn1 but not Ft-L and TfR1 and significantly enhanced ERK phosphorylation. Furthermore, the pretreatment of cultured hepatocytes with either a pERK inhibitor or a GHSR1α antagonist abolished the effects of ghrelin on Fpn1 expression and ERK phosphorylation. Our findings confirmed that fasting increases iron export in the liver by upregulating Fpn1 expression through the ghrelin/GHSR1α/MAPK signaling pathway.

Rosuvastatin improves olanzapine’s effects on behavioral impairment and hippocampal, hepatic and metabolic damages in isolated reared male rats

Background & aimSchizophrenia is a chronic, disabling neurological illness. This study investigated the effect of rosuvastatin (RSU) addition to the antipsychotic drug: olanzapine (OLZ) in treatment of post-weaning isolation rearing (IR) damaging effect and assessed behavioral impairment, metabolic and hepatic abnormalities, oxidative stress, and inflammatory markers. MethodsTreatment with OLZ (6 mg/kg, P.O.) and/or RSU (10 mg/kg, I.P.) have been started 6 weeks after isolation. We assessed behavioral tests, serum cortisol level, and hippocampal content of neurotransmitters. In addition, we assessed histopathology, inflammatory and oxidative stress markers of hippocampus, liver and adipose tissue ResultsTreatment of IR animals with OLZ, and/or RSU significantly counteracted the changes in hippocampus, liver and adipose tissue induced by post-weaning IR. Co-treatment of IR rats with both OLZ and RSU showed additive effects in some areas like improving both tumor necrosis factor alpha (TNFα) in both hippocampus and liver, histopathology of liver, oxidative stress markers of adipose tissue, β3 adrenergic receptors (ADRβ3), serum cortisol and total cholesterol. In addition, RSU alone alleviated the damage of IR rats by the same efficacy as OLZ with more benefit in cognition and exploration. Conclusionpost-weaning IR as a model has behavioral, hippocampal, hepatic and marked metabolic changes more relevant to schizophrenia than drug-induced models. These effects were ameliorated by RSU and/or OLZ that are explained by their antioxidant, anti-inflammatory, anti-stress and anti-hyperlipidemic properties. Interestingly, co-treatment with both drugs showed a better effect.

EJE PRIZE 2018: A gut feeling about glucagon.

Hyperglucagonaemia (in the fasting as well as in the postprandial state) is considered a core pathophysiological component of diabetes and is found to contribute substantially to the hyperglycaemic state of diabetes. Hyperglucagonaemia is usually viewed upon as a consequence of pancreatic alpha cell insensitivity to the glucagon-suppressive effects of glucose and insulin. Since we observed that the well-known hyperglucagonaemic response to oral glucose in patients with type 2 diabetes is exchanged by normal suppression of plasma glucagon levels following isoglycaemic intravenous glucose administration in these patients, we have been focusing on the gut and gut-derived factors as potential mediators of diabetic hyperglucagonaemia. In a series of clinical experiments, we have elucidated the role of gut-derived factors in diabetic hyperglucagonaemia and shown that glucose-dependent insulinotropic polypeptide promotes hyperglucagonaemia and that glucagon, hitherto considered a pancreas-specific hormone, may also be secreted from extrapancreatic tissues - most likely from proglucagon-producing enteroendocrine cells. Furthermore, our observation that fasting hyperglucagonaemia is unrelated to the diabetic state, but strongly correlates with obesity, liver fat content and circulating amino acids, has made us question the common 'pancreacentric' and 'glucocentric' understanding of hyperglucagonaemia and led to the hypothesis that steatosis-induced hepatic glucagon resistance (and reduced amino acid turnover) and compensatory glucagon secretion mediated by increased circulating amino acids constitute a complete endocrine feedback system: the liver-alpha cell axis. This article summarises the physiological regulation of glucagon secretion in humans and considers new findings suggesting that the liver and the gut play key roles in determining fasting and postabsorptive circulating glucagon levels.

Effects of replacing wheat flour with detoxified ginkgo nut powder on lipid metabolism of obese C57BL/6J male mice

ABSTRACT Effects of detoxified ginkgo nut powder (DGP) on lipid metabolism of obese mice were evaluated. Mice were divided into four groups: normal group (NC), hyperlipidaemic group (HC, high-fat feed), HC/DGP group (high-fat diet + DGP), and HC/flour group (HC/WF, high-fat diet + flour). After 18 weeks, liver and body fat weights of HC/DGP were significantly lower than those of HC and HC/WF, but similar to those of NC. The serum and hepatic levels of serum triglyceride, total cholesterol, low-density lipoprotein cholesterol, leptin and resistin in HC/DGP were similar to those in NC but significantly lower than those in HC and HC/WF. In addition, the expression levels of PPARα (peroxisome proliferator-activated receptor alpha) in liver and of hormone-sensitive lipase, adipose triglyceride lipase, and monoacylglycerol lipase in epididymal fat were higher in HC/DGP than in other groups. The results revealed that DGP may reduce inflammation and promote normal lipid metabolism.

Supplementation of Mussel Peptides Reduces aging Phenotype, Lipid Deposition and Oxidative Stress in D-Galactose-Induce Aging Mice.

Aging is associated with glucose and lipid metabolism disorder. We aimed to examine the effects of mussel peptides on protecting against aging by regulating glucose and lipid metabolism. For the aging model, d-galactose (200 mg/kg) was administered subcutaneously to 8-month-old mice for 8 weeks. Mussel peptides (1,000 mg/kg) were simultaneously administered by intragastric gavage. The glucose and lipid metabolism profiles, aging phenotype and peroxisome proliferator-activated receptors (PPARs) expression in the liver and adipose tissue of ICR mice were measured. D-galactose-treated mice showed pronounced fat deposition and impaired glucose and lipid homeostasis, along with increased oxidative damage and aging. Mussel peptides improved metabolic status by reducing serum glucose and triglyceride levels, insulin resistance and hepatic free fatty acid, as well as enhancing serum high-density lipoprotein (HDL) level and hepatic glycogen content, accompanied with amelioration of aging phenotype and fat deposition. Moreover, mussel peptides ameliorated oxidative stress in aged liver tissues and promoted expression of peroxisome proliferator activated receptors alpha (PPARα) and gamma (PPARγ) in liver and adipose tissues. These results indicate that mussel peptides protect against lipid metabolic disorders associated with aging via maintaining oxidative stress homeostasis and elevated expression levels of PPARs.

Esculin attenuates endotoxin shock induced by lipopolysaccharide in mouse and NO production in vitro through inhibition of NF-κB activation

Esculin, a coumarin compound derived from the traditional Chinese herbs such as Cortex Fraxini, has long been used for treating inflammatory and vascular diseases. In present study, we analyzed the role of esculin against macrophages and endotoxin shock induced by lipopolysaccharide (LPS) in mice. Here, we demonstrated that esculin suppressed inflammatory reactions in macrophages and protected mice from LPS-induced endotoxin shock. We found that esculin significantly inhibited the production of nitric oxide (NO) production via the inhibition of nuclear factor-κB (NF-κB) activation in macrophages. In animal model, esculin pretreatment significantly improved the survival rate of mice. LPS-induced increase of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) in serum, lung, liver and kidney were markedly inhibited by esculin. IL-10, an anti-inflammatory cytokine, was up-regulated by esculin. Moreover, the histopathological analyses showed that esculin significantly attenuated the tissues injury of lung, liver, kidney in endotoxic mice. In addition, esculin significantly diminished the protein expression of NF-κB p65 in lung, liver, kidney, which resulted in lower levels of inflammatory mediators. These results suggest that esculin may be a potential drug for treatment of various inflammatory diseases.

Thyroid hormone regulates vitellogenin by inducing estrogen receptor alpha in the goldfish liver

Vitellogenin (Vtg) is an egg-yolk precursor protein that is synthesized in the liver of oviparous species and taken up from the circulation by the ovary. It is well known that Vtg is induced by circulating estrogens. However, other endocrine factors that regulate the expression of Vtg are less well characterized; factors that might play significant roles, especially in seasonal spawners such as the goldfish which require increased quantities of Vtg for the development of hundreds of follicles. In this regard, thyroid hormones have been shown to cycle with the reproductive season. Therefore, we hypothesized that the thyroid hormones might influence the synthesis of Vtg. Treatment of female goldfish with triiodothyronine (T3) resulted in increased Vtg, an observation that was absent in males. Furthermore, T3 failed to induce Vtg in cultured hepatocytes of either sex. Interestingly however, T3 consistently up-regulated the expression of the estrogen receptor alpha (ERα). The T3 mediated upregulation of ERα requires the presence of both thyroid receptor (TR) α-1 and TRβ. When goldfish or cultured hepatocytes were treated with T3 followed by estradiol, there was a synergistic increase in Vtg, a response which is dependent on the presence of ERα. Therefore, by upregulating ERα, T3 serves to prime the liver to subsequent stimuli from estradiol. This cross-talk likely reveals an important physiologic mechanism by which thyroid hormones, whose circulating levels are high during early gonadal recrudescence, facilitate the production of large amounts of Vtg required for egg development.

Abstract 1039: Metabolic hormones FGF19/15 promote hepatocellular carcinoma through activation of CSCs in fatty liver

Abstract Background: Fibroblast growth factor 19 (FGF19), a member of the fibroblast growth factor (FGF) family, is a hormone that regulate glucose, lipid, and energy homeostasis in human. It has been shown that FGF19 and its specific receptor FGFR4 play an important role in non-alcoholic steatohepatitis (NASH) which is closely related hepatocellular carcinoma (HCC). Recently, we found FGF19 overexpressed in the HCC tissue and serum in patients. The high level of FGF19 was positively related to cancer stem cells (CSCs) specific surface markers such as CD133 and EpCAM. The aim of this study is to identify potential mechanism in which FGF15 (the FGF19's orthologous protein in mouse)/FGFR4 associated abnormal lipid metabolism initiates HCC progression in a NASH-HCC mouse Model. Methods: In vivo study, male C57L/J mice were injected intraperitoneally with N-nitrosodiethylamine(DEN) when 2 weeks old. Both the DEN and control mice were divided into two groups based on receiving high fat diet (HFD) or control diet (CD). Tumor nodules in the liver were monitored by ultrasound. Body weight, glucose tolerance test (GTT) and insulin tolerance test (ITT) were recorded during the experimental period. The liver weights, tumor volume, histology, alanine transaminase (ALT), alpha fetoprotein (AFP), and triglyceride (TG) were determined at 8, 20 and 32weeks. In vitro study, human HCC cell lines (Hep3B and Huh7) as well as mouse HCC cell lines (Hepa1-6 and FL83B,) were exposed to a long-chain mixture of free fatty acid(FFA) at different concentrations (UT, 0.5mM and 1mM FFA) for 24 and 48 hours, respectively. EpCAM and CD133 were determined by flow cytometery in the isolated tumor cells from HCC tissues. FGF19/15, FGFR4, CSCs surface markers and Wnt signaling pathway (β-catenin, GSK3β) were analyzed by RT-PCR and Western blot. FGFR4 signal was blocked to elucidate the role of FGF19/15 during the CSCs activation in vitro. In addition, FGF19, FGFR4 and EpCAM were also analyzed in human HCC samples. Results: Metabolic disorders were observed when DEN mice fed with HFD. HCC incidence and tumor volume were significantly increased in DEN+HFD group compared to that in DEN+CD group. DEN mice fed with HFD showed increased levels of FGF15/FGFR4 and β-catenin as well as the CSCs surface makers. Increased expression of CSCs surface makers were also found in the HCC cell lines when exposure to FFA in vitro. Blockage the FGF19 signal can abolish Wnt/β-catenin pathway and cell stemness features in the FFA environment. Increased levels of FGF19, FGFR4 and EpCAM were found in human HCC samples. Conclusion: Up-regulated FGF19/15 and FGFR4 promoted the development of HCC by active CSCs signaling in metabolic disorder microenvironment. FFA enhanced the CSCs activation was partially dependent on Wnt/β-catenin signaling. FGF19/FGFR4 could be a potential therapeutic target in HCC patients. Citation Format: Guozhen Cui, Xingkai Liu, Harshul Pandit, Yingbin Yang, Suping Li, Lu Cai, Robert C Martin, Hengjun Zhao, Wei Li, Yan Li. Metabolic hormones FGF19/15 promote hepatocellular carcinoma through activation of CSCs in fatty liver. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1039.

Dietary triacylglycerols with palmitic acid in the sn-2 position modulate levels of N-acylethanolamides in rat tissues.

BackgroundSeveral evidences suggest that the position of palmitic acid (PA) in dietary triacylglycerol (TAG) influences different biological functions. We aimed at evaluating whether dietary fat with highly enriched (87%) PA in sn-2 position (Hsn-2 PA), by increasing PA incorporation into tissue phospholipids (PL), modifies fatty acid profile and biosynthesis of fatty acid—derived bioactive lipids, such as endocannabinoids and their congeners.Study DesignRats were fed for 5 weeks diets containing Hsn-2 PA or fat with PA randomly distributed in TAG with 18.8% PA in sn-2 position (Lsn-2 PA), and similar total PA concentration. Fatty acid profile in different lipid fractions, endocannabinoids and congeners were measured in intestine, liver, visceral adipose tissue, muscle and brain.ResultsRats on Hsn-2 PA diet had lower levels of anandamide with concomitant increase of its congener palmitoylethanolamide and its precursor PA into visceral adipose tissue phospholipids. In addition, we found an increase of oleoylethanolamide, an avid PPAR alpha ligand, in liver, muscle and brain, associated to higher levels of its precursor oleic acid in liver and muscle, probably derived by elongation and further delta 9 desaturation of PA. Changes in endocannabinoids and congeners were associated to a decrease of circulating TNF alpha after LPS challenge, and to an improved feed efficiency.ConclusionsDietary Hsn-2 PA, by modifying endocannabinoids and congeners biosynthesis in different tissues may potentially concur in the physiological regulation of energy metabolism, brain function and body fat distribution.

Soybean Flour Improves Fatty Acid Profile and Decreases Hepatic Triglyceride Deposition in Rats Fed with Normocaloric and Hypercaloric Diet

This study investigated the effects of replacing casein with soy flour on the fatty acids profile and triglycerides metabolism in the liver of rats that were previously fed with normocaloric and hypercaloric diets based on casein. Wistar male rats were used; one group was fed with control diet (AIN-93) and another with hypercaloric diet (AIN-93 with 34.15% sucrose, 42% fat calories) for 9 weeks. Each group was then divided into two subgroups and casein was replaced with soybean in one of them, obtaining CC (control casein), CS (control soy), HC (hypercaloric casein) and HS (hypercaloric soy), which were fed for 6 weeks. We measured triglycerides in serum, and triglycerides, total lipids, fatty acids profile, the expression of apolipoprotein B (Apo B), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), sterol-regulatory element-binding protein 1c (SREBP-1c), mitochondrial glycerol-3-phosphate acyltransferase (mGPAT), diacylglycerol acyltransferase 2 (DGAT-2), carnitine palmitoyltransferase 1 (CPT-1) and peroxisome proliferator-activated receptors alpha (PPARα) in liver. Histological studies were also performed. When comparing HS vs. HC, a positive effect of soybean flour on hepatic triglycerides deposits was found, possibly through the reduction in DGAT-2 expression (P < 0.01) and the increase in Apo B (P < 0.001) expression. Soybean flour also decreased fat deposits in control diets when compared with casein, decreasing the DGAT-2 (P < 0.001) expression and increasing Apo B (P < 0.001), CPT-1 (P < 0.05) and PPARα (P < 0.01) expressions. Both soy diet subgroups increased unsaturated fatty acids respect to casein diets (P < 0.01). Hepatocytes showed few lipid droplets in HS, whereas a fat deposit in HC was observed. These results suggest that replacing casein with soybean flour in normocaloric and hypercaloric diets reduces triglycerides and improves fatty acids profile in rat liver.

Biological functional annotation of retinoic acid alpha and beta in mouse liver based on genome-wide binding.

Retinoic acid (RA) has diverse biological effects. The liver stores vitamin A, generates RA, and expresses receptors for RA. The current study examines the hepatic binding profile of two RA receptor isoforms, RARA (RARα) and RARB (RARβ), in response to RA treatment in mouse livers. Our data uncovered 35,521, and 14,968 genomic bindings for RARA and RARB, respectively. Each expressed unique and common bindings, implying their redundant and specific roles. RARB has higher RA responsiveness than RARB. RA treatment generated 18,821 novel RARB bindings but only 14,798 of RARA bindings, compared with the control group. RAR frequently bound the consensus hormone response element [HRE; (A/G)G(G/T)TCA], which often contained the motifs assigned to SP1, GABPA, and FOXA2, suggesting potential interactions between those transcriptional factors. Functional annotation coupled with principle component analysis revealed that the function of RAR target genes were motif dependent. Taken together, the cistrome of RARA and RARB revealed their extensive biological roles in the mouse liver. RAR target genes are enriched in various biological processes. The hepatic RAR genome-wide binding data can help us understand the global molecular mechanisms underlying RAR and RA-mediated gene and pathway regulation.

Lipid-lowering effects of Danhong injection on hyperlipidemia rats

Ethnopharmacological relevanceDanhong injection (DHI), a Chinese medical product extracted from Radix et Rhizoma Salviae Miltiorrhizae (Salvia miltiorrhiza Bge., Labiatae, Danshen in Chinese) and Flos Carthami (Carthamus tinctorius L., Compositae, Honghua in Chinese), has been reported to have anti-inflammatory, anti-oxidative and anti-fibrinolytic properties and is used extensively for the clinical treatment of cardiovascular disease in clinic. This study aimed to investigate the preventive and therapeutic effects of DHI on hyperlipidemia. Materials and methodsForty-eight adult male Sprague-Dawley rats were randomly divided into four groups: normal control (NC), model control (MC) and DHI-treated at doses of 1.0mL/kg and 2.0mL/kg. The effects of DHI on serum triglyceride (TG), total cholesterol (TC), glucose, high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) were evaluated and insulin was determined by enzyme-linked immunosorbent assay (ELISA). Moreover, the expression of acetyl-CoA carboxylase 1 (ACC1), fatty acid synthase (FAS), carnitine palmitoyl transferase 1 (CPT1), hydroxymethylglutaryl-CoA reductase (HMGR) and peroxisome proliferator-activated receptor alpha (PPAR-α) in liver were determined by real-time PCR. ResultsCompared with the MC group, rats treated with DHI had significantly reduced TG, TC, LDL-C and arteriosclerosis index (AI). Expression of FAS and HMGR mRNA was significantly reduced, whereas the CPT1 and PPAR-α were significantly increased. ConclusionDHI treatment was accompanied by significantly increased lipolysis in the liver and decreased fatty acid synthesis. The insights gained from this study will improve both understanding of the mechanisms involved in the effect of DHI on hyperlipidemia and the pharmacological rationale for the use of DHI in diseases caused by lipid metabolic disorders.

Effect of garlic on lipid profile and expression of LXR alpha in intestine and liver of hypercholesterolemic mice

BackgroundGarlic is one of the medicinal plants which has showed beneficial effects on atherosclerosis risk factors. The liver X receptor α (LXRα) is an important regulator of cholesterol, triglyceride and glucose homeostasis that belongs to the nuclear receptor superfamily. In this study we investigated the effect of garlic on lipid profile, glucose as well as LXRα expression in intestine and liver of mice.MethodsForty male N-Mary mice were randomly divided into 3 groups (n = 8): group1 received chow + 2% cholesterol + 0.5% cholic acid, group 2: chow + 4% (w/w) garlic extract + 2% cholesterol + 0.5% cholic acid, and group 3: chow only. After one month of treatment, mice were anesthetized, blood was collected from their heart, and the first 10 cm of the small intestine and liver were removed. Glucose was measured by a glucometer; other biochemical factors were measured by enzymatic methods. LXR expression was checked by RT-PCR and western blotting.ResultsCompared with hypercholesterolemic mice, treatment with garlic extract significantly decreased total cholesterol, low-density lipoprotein cholesterol (LDL-C), triglycerides, very low density lipoprotein-cholesterol (VLDL-C), atherogenic index, alanine aminotranferease (ALT) and aspartate aminotransferase (AST) (all of them P < 0.05). Change in HDL-C levels was not significant in garlic-extract treated animals compared with hypercholesterolemic group. LXR protein and mRNA in the intestine were increased in garlic-extract treated group compared with chow group (P < 0.05), while in the liver, only mRNA of LXR was increased in hypercholesterolemic control mice (P < 0.05).ConclusionsThe present study demonstrated that garlic extract reduced LXRα expression in the liver and increased its expression in the intestine. These effects probably have an important role in reducing serum triglyceride and cholesterol.