PPARα mRNA Research Articles

The roles and interaction of FXR and PPARs in the pathogenesis of nonalcoholic fatty liver disease

Background and study aimsTo identify the roles and interaction of farnesoid X receptor (FXR) and peroxisome proliferator activated receptors (PPARs) in Non-alcoholic fatty liver disease (NAFLD) pathogenesis. Material and Methods16 C57/BL male FXR knockout (KO) mice and sex- and age-matched C57/BL wild type mice were received either standard rodent chow or high-fat and sucrose diet (Blank control, NAFLD, FXR KO and FXR KO NAFLD) for 8 weeks. After that, all mice were sacrificed. Liver tissues and blood samples were collected for laboratory and RT-PCR examination. ResultsNAFLD, FXR KO and FXR KO NAFLD mouse models were successful established. Compared with blank control, FXR and PPAR-α mRNA expression decreased significantly (P < 0.05), PPAR-β expression increased slightly (P > 0.05), PPAR-γ expression increased significantly in NAFLD (P < 0.05). Slight increased PPAR-α mRNA expression (P > 0.05) and markedly decreased PPAR-β and PPAR-γ expression (P < 0.05) were found in FXR KO. Compared with FXR KO group, there was a slight increase in PPAR-αand PPAR-βmRNA expression (P > 0.05) and significant increase in PPAR-γ expression (P < 0.05) in FXR KO NAFLD group. Comparison with NAFLD, PPAR-α mRNA expression increased slightly (P > 0.05), PPAR-β and PPAR-γ expression decreased significantly (P < 0.05) in FXR KO NAFLD. ConclusionFXR and PPARs interaction may play important roles in NAFLD pathogenesis.

Sex-dependent metabolic effects of pregestational exercise on prenatally stressed mice.

Stressful events during the prenatal period have been related to hyperactive hypothalamic-pituitary-adrenal (HPA) axis responses as well as metabolic changes in adult life. Moreover, regular exercise may contribute to the improvement of the symptoms associated with stress and stress-related chronic diseases. Therefore, this study aims to investigate the effects of exercise, before the gestation period, on the metabolic changes induced by prenatal stress in adult mice. Female Balb/c mice were divided into three groups: control (CON), prenatal restraint stress (PNS) and exercise before the gestational period plus PNS (EX+PNS). When adults, the plasmatic biochemical analysis, oxidative stress, gene expression of metabolic-related receptors and sex differences were assessed in the offspring. Prenatal stress decreased neonatal and adult body weight when compared to the pregestational exercise group. Moreover, prenatal stress was associated with reduced body weight in adult males. PNS and EX+PNS females showed decreased hepatic catalase. Pregestational exercise prevented the stress-induced cholesterol increase in females but did not prevent the liver mRNA expression reduction on the peroxisome proliferator-activated receptors (PPARs) α and γ in PNS females. Conversely, PNS and EX+PNS males showed an increased PPARα mRNA expression. In conclusion, pregestational exercise prevented some effects of prenatal stress on metabolic markers in a sex-specific manner.

Investigation of the Lipid-Lowering Mechanisms and Active Ingredients of Danhe Granule on Hyperlipidemia Based on Systems Pharmacology.

ObjectiveInvestigate the active ingredients and underlying hypolipidemic mechanisms of Danhe granule (DHG).MethodsThe lipid-lowering effect of DHG was evaluated in hyperlipidemic hamsters induced by a high-fat diet. The ingredients absorbed into the blood after oral administration of DHG in hamsters were identified by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS). A systems pharmacology approach incorporating target prediction and network construction, gene ontology (GO) enrichment and pathway analysis was performed to predict the active compounds and map the compounds-targets-disease network. Real-time polymerase chain reaction (RT-PCR) and Western blot were utilized to analyze the mRNA and protein expression levels of predicted targets.ResultsDHG remarkably lowered the levels of serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-c), and arteriosclerosis index (AI), at the same time, elevated the levels of serum high-density lipoprotein cholesterol (HDL-c) and HDL-c/TC ratio in hyperlipidemic hamsters. Sixteen ingredients absorbed into blood after oral administration of DHG were identified as the possible components interacted with targets. Moreover, 65 potential targets were predicted after targets intersection and compounds–targets–disease network mapping. Then, compounds–targets–pathways network mapping revealed that six active compounds (emodin, naringenin, etc.) compounds could interact with 10 targets such as sterol regulatory element binding protein (SREBP) 1c, SREBP-2 and peroxisome proliferation-activated receptor (PPAR) α, regulate three lipid metabolism-related pathways including SREBP control of lipid synthesis pathway, PPAR signaling pathway and nuclear receptors in lipid metabolism and toxicity pathway, and further affect lipid metabolic processes including fatty acid biosynthesis, low-density lipoprotein receptor (LDLR)-mediated cholesterol uptake, bile acid biosynthesis, and cholesterol efflux. Experimental results indicated that DHG significantly increased SREBP-2, LDLR, PPARα, liver X receptor alpha (LXRα), cholesterol 7α-hydroxylase (CYP7A1), and ATP binding cassette subfamily A member 1 (ABCA1) mRNA and protein expressions while decreased SREBP-1c and fatty acid synthase (FAS) mRNA, and protein expressions.ConclusionDHG possessed a good hypolipidemic effect that may be through affecting the mRNA and protein expressions of SREBP-1c, FAS, SREBP-2, LDLR, PPARα, LXRα, CYP7A1, and ABCA1, involving in fatty acid synthesis, LDLR-mediated cholesterol uptake, bile acid biosynthesis, and cholesterol efflux. This study further provided experimental evidence about its practical application for treating hyperlipidemia and its complications.

Effect of partial black soldier fly (Hermetia illucens L.) larvae meal replacement of fish meal in practical diets on the growth, digestive enzyme and related gene expression for rice field eel (Monopterus albus)

A ten-week feeding trial on Monopterus albus (initial weight 24.02 ± 0.02 g) was conducted to evaluate effects of four isonitrogenous and isoenergetic diets in which different proportions of fish meal (0, 2.5 %, 5% and 7.5 %) were substituted with Hermetia illucens (HI) larvae meal, corresponding to dietary inclusion levels of 0 (FM), 5.26 % (HI5.26), 10.52 % (HI10.52) and 15.78 % (HI15.78). Compared with FM, weight gain rate and condition factor of M. albus and gumminess and adhesiveness of its muscle tissue were significantly increased in HI5.26 and HI10.52 groups (P < 0.05); whereas feed conversion ratio was markedly decreased in HI5.26 (P < 0.05), and expression of hepatic FASD2 and ME2 was significantly upregulated in HI5.26 (P < 0.05). Compared with FM, cohesiveness of muscle was significantly decreased in HI groups, but hardness of muscle and expression of hepatic FAS were remarkably upregulated in HI groups (P < 0.05); ACC and ME1 mRNA levels were significantly upregulated in HI10.52 (P < 0.05); CPT2 mRNA levels were significantly upregulated in HI5.26 and HI15.78 (P < 0.05); PPARγ, G6PD and HSL mRNA levels were significantly upregulated in HI10.52 and HI15.78 (P < 0.05). Springiness and chewiness of muscle, activity of intestinal lipasewere, serum indexes (glutamic oxalacetic transaminase, catalase, malondialdehyde), several gut bacterial indices (shannon, chao1, ACE, OTUs abundance of proteobacterias) and abundance of hepatic PPARα mRNA in HI15.78 were remarkably lower than those in FM (P < 0.05), while content of crude lipid, gut firmicutes and hepatic Dgat2 mRNA were significantly lower than FM (P < 0.05). Results showed that HI improved growth performance and balance of gut bacteria of M. albus when supplemented at a rate below 15.78 %, but negative effects on lipid metabolism were observed when HI was supplemented at excessive levels.

Decreased Expression of Peroxisome Proliferator-activated Receptor α Gene as an Indicator of Metabolic Disorders in Stunting Toddler

BACKGROUND: Stunting in children increases the risk of degenerative diseases in adulthood, including dyslipidemia, obesity, type 2 diabetes mellitus, and cardiovascular disease. This is based on the result of metabolic changes that may be caused by chronic malnutrition and experienced by stunting children. Stunting in children is associated with metabolic disorders that are based on impaired fat oxidation, a trigger factor for obesity in adulthood. The peroxisome proliferator-activated receptor (PPAR) α gene is a transcriptional factor that regulates fat, carbohydrate, and amino acid metabolism whose genetic variants are linked to the development of dyslipidemia and cardiovascular disease.&#x0D; AIM: The study assessed the effect of metabolic changes in stunting toddler on PPARα gene expression.&#x0D; MATERIALS AND METHODS: An analytical-observational laboratory was done using 41 blood samples, coming from 23 stunting toddlers, and 18 not-stunting toddlers. In all research subjects, anthropometric measurements and examination of PPARα gene mRNA expression were carried out. Analysis of PPARα gene mRNA expression using one-step quantitative reverse transcriptase-polymerase chain reaction using specific primers, as a comparison of gene expression using the GAPDH gene. The relative expression of the PPARα mRNA gene was analyzed using the LIVAK formula.&#x0D; RESULTS: The study obtained a mean of ΔCT in stunting toddlers of 5.81, whereas in stunting toddlers at 5.082. Analysis with LIVAK 2 ^ - formula (ΔCT stunting -ΔCT not stunting) obtained PPARα mRNA gene expression of 0.6.&#x0D; CONCLUSION: We conclude that there is a decrease in PPARα gene expression in stunting toddlers.

Repressor Activator Protein 1 Worsens Cardiomyopathy in Diet‐induced Type 2 Diabetic Mice

BackgroundDiabetic cardiomyopathy is defined by the structural and functional abnormalities of the myocardium in the absence of other cardiac risk factors in diabetic patients. In diabetes, the increment of fatty acid metabolism and subsequently cardiac inefficiency and myocardial lipid accumulation may underlie the pathology of diabetic cardiomyopathy. The telomeric protein repressor activator protein 1 (Rap1) is an identified regulator that regulates gene transcription by binding to non‐telomeric sites. In particular, Rap1 can bind to nuclear PPARα (an important transcriptional regulator of lipid metabolism) loci, and regulate transcription of PPARα and its target genes, including CD36 (fatty acid transporter). However, the effects of Rap1 on diabetic cardiomyopathy are largely unknown. Thus, the present study investigated if Rap1 deficiency protects against diabetic cardiomyopathy and explored the underlying mechanisms.Methods and ResultsHigh fat diet feeding is a widely used protocol to induce type 2 diabetes in mice. Upon high fat feeding (60% fat) for 12 weeks, male Rap1 knockout (KO) mice displayed significantly reduced body weight gain compared with their wild type (WT) littermates. The glucose tolerance test and insulin tolerance test showed that Rap1 deficiency in the high fat diet fed‐mice led to improved glucose intolerance and insulin action. Furthermore, Rap1 deficiency attenuates high fat diet‐induced cardiac structural remodeling and dysfunction in mice, as evidenced by significantly reduction in left ventricular internal dimension in systole (LVIDs), left ventricular mass, increment in ejection fraction (EF) and fractional shortening (FS), as well as improving of myocardial performance index (MPI). These changes were associated with remarkable reductions in the mRNA and protein expression of PPARα and CD36 in the myocardium of high fat fed‐Rap1 KO mice, indicating that Rap1 deficiency may protect against diabetic cardiomyopathy via the suppression of PPARα/CD36 signaling.ConclusionIn summary, these results suggest that Rap1 impairment has cardioprotective consequences in the high fat diet‐induced diabetes model, identifying a new dimension in the roles of endogenous Rap1.Support or Funding InformationThe authors’ work was supported by The General Research Fund (17117217M, 17123718M, Research Grants Council of Hong Kong) and The National Natural Science Foundation of China (No. 81800245).

The effect and mechanism of dietary monocalcium phosphate restriction on hepatic lipid deposition and immune status in obscure puffer, Takifugu obscurus

A 12-week chronic feeding trial was conducted to determine the effect and mechanism of dietary monocalcium phosphate (CaH2PO4) restriction on hepatic lipid deposition and immune status in obscure puffer, Takifugu obscurus. Fish with an initial mean weight of 7.21 ± 0.05 g were fed a phosphorus (P)-sufficient diet (P6, diet supplemented with 0.6% P) or P-deficient diet (P0, diet supplemented without P). Sampling occurred at weeks 3, 6, 9 and 12, respectively. During the feeding trial, fish fed the P0 diet showed poor growth performance, reduced feed intake, whole body P and ash as well as vertebrae P contents, which are signs of P deficiency. Dietary P restriction increased hepatic vacuoles and lipid droplets, lipid content of whole body and liver, viscerosomatic and hepatosomatic indices, whereas reduced whole body protein content and lipolytic enzyme activities, including carnitine palmitoyltransferase 1 (CPT-1) and lipoprotein lipase (LPL). But peroxisome proliferators-activated receptor α (pparα) mRNA level was up-regulated by dietary P deficiency on week 3, and then down-regulated on weeks 6, 9 and 12. Meanwhile, dietary P deficiency increased hepatic mRNA expressions of 6-phosphogluconate dehydrogenase (6pgd), malic enzyme (me), acetyl-CoA carboxylase (acc)-α and acc-β, while decreased cpt-1 and fatty acid binding protein (fabp). Additionally, dietary P deficiency decreased the mRNA level of complement 3 (c3) while increased heat shock proteins (HSPs, including hsp90 and hsp70) on weeks 9 and 12. Dietary P restriction also induced apoptosis by up-regulating the mRNA levels of caspase-3, caspase-8 and caspase-9 and down-regulating the levels of B-cell lymphoma 2 (bcl-2) and Bax inhibitor-1 (bi-1). In conclusion, our study indicated that dietary P deficiency induced lipid deposition and impaired the immune status of obscure puffer.

Effects of 1,25(OH)2 D3 on lipid droplet growth in adipocytes.

This study aimed to explore the effects of 1,25(OH)2 D3 on lipid droplet (LD) growth in 3T3-L1 adipocytes of hypertrophy model. Cocktail method was used to induce differentiation in 3T3-L1 cells. After 8 days, the cells were modeled by 100, 300, 600, and 900 μM palmitic acid (PA) for 24 hr. The best concentration of modeling was screened by MTT results and triglycerides (TG) content. The model cells were intervened by 1, 10, and 100 nM 1,25(OH)2 D3 for 24 hr. Then, the TG content of cells were detected and stained by oil red O. The diameter and quantity of LDs were analyzed. mRNA relative expression levels of genes related to LD (CIDE-a, Fsp27, PLIN-1), upstream response factor (PPAR-α, PPAR-γ, and VDR), and TG metabolism (long chain acyl-CoA synthetase 3, 1-acylglycerol-3-phosphate O-acyltransferase 1, adipose triglyceride lipase, diacylglycerol acyltransferase 1, diacylglycerol acyltransferase 2, glycerol-3-phosphate O-acyltransferase 3, glycerol-3-phosphate O-acyltransferase 4, hormone-sensitive lipase, mannosyl (alpha-1,3-)-glycoprotein beta-1,2-N-acetyl glucosaminyl transferase, phosphatidic acid phosphatase, and uncoupling protein-1) were detected by RT-qPCR. A total of 300 μM PA was selected as the optimum concentration. Compared with model group, 10 and 100 nM 1,25(OH)2 D3 decreased the average diameter, increased the quantity of LDs, upregulated PPAR-α and PLIN-1 mRNA expression levels, and downregulated CIDE-a and Fsp27 mRNA expression levels significantly (p < .05). However, 1 nM 1,25(OH)2 D3 did not alter LD morphology and TG content. mRNA expression levels of long chain acyl-CoA synthetase 3, 1-acylglycerol-3-phosphate O-acyltransferase 1, diacylglycerol acyltransferase 2, glycerol-3-phosphate O-acyltransferase 3, and glycerol-3-phosphate O-acyltransferase 4 in 10 and 100 nM groups were significantly lower than those in the model group (p < .05); mRNA expression levels of adipose triglyceride lipase, diacylglycerol acyltransferase 1, hormone-sensitive lipase, mannosyl (alpha-1,3-)-glycoprotein beta-1,2-N-acetyl glucosaminyl transferase, phosphatidic acid phosphatase, and uncoupling protein-1 were significantly increased in the 100 nM group (p < .05). The 10 and 100 nM 1,25(OH)2 D3 can inhibit LD fusion, promote LD decomposition, reduce LD volume, and inhibit lipogenesis through the PPAR-α signaling pathway.

Mogroside V Protects against Hepatic Steatosis in Mice on a High-Fat Diet and LO2 Cells Treated with Free Fatty Acids via AMPK Activation.

Previous studies presented various beneficial effects of mogrosides extract from Siraitia grosvenorii, which has been included in the list of Medicine Food Homology Species in China. Mogroside V (MV) is one of the main ingredients in mogrosides extract; however, whether and how MV improves impaired lipid metabolism in the liver remains to be elucidated. Herein, we investigated the therapeutic effects of mogroside V upon hepatic steatosis in vivo and in vitro and explored the underlying mechanisms. The results showed that MV significantly ameliorated hepatic steatosis in high-fat diet- (HFD-) fed mice. Furthermore, the increased protein expression of PPAR-γ, SREBP-1, and FASN and mRNA expression of pparg, srebp1, scd1, and fasn in the liver in HFD-fed mice, which contribute to de novo lipogenesis, were dose-dependently reversed by MV treatment. Meanwhile, MV counteracted the suppressed expression of PPAR-α and CPT-1A and mRNA expression of atgl, hsl, ppara, and cpt1a, thus increasing lipolysis and fatty acid oxidation. In addition, in free fatty acids- (FFAs-) incubated LO2 cells MV downregulated de novo lipogenesis and upregulated lipolysis and fatty acid oxidation, thereby attenuating lipid accumulation, which was significantly abrogated by treatment with Compound C, an inhibitor of AMP-activated protein kinase (AMPK). Taken together, these results suggested that MV exerted a pronounced effect upon improving hepatic steatosis through regulating the disequilibrium of lipid metabolism in the liver via an AMPK-dependent pathway, providing a potential lead compound candidate for preventing nonalcoholic fatty liver disease.

Anti-obesity effects of individual or combination treatment with Spirulina platensis and green coffee bean aqueous extracts in high-fat diet-induced obese rats

Background: Obesity is associated with serious health risks, including premature mortality. This study investigated the anti-obesity effects of individual or combination treatment with Spirulina platensis and green coffee bean aqueous extracts in high-fat diet-induced (HFD) obese rats.Methods: Rats were fed on HFD to induce obesity. Corn oil in the HFD accounted for 50.98% of the calories. Fifty rats were divided into five groups (10 rats/group): control, HFD, HFD-Spirulina, HFD-coffee, HFD-Spirulina and coffee cotreatment groups. The serum levels of lipid, leptin, and insulin, as well as the hepatic mRNA levels of fatty acid synthase (FAS), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α), and peroxisome proliferator-activated alpha receptor (PPARα) were estimated.Results: The Spirulina and/or green coffee bean aqueous extracts decreased the final bodyweight and liver weight, and the serum levels of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase. The extracts decreased the serum levels of total cholesterol, triacylglycerol, low-density lipoprotein-cholesterol, leptin, and glucose, and enhanced the serum insulin level. Furthermore, the extracts enhanced the PGC-1α and PPARα mRNA levels and decreased the FAS mRNA levels.Conclusion: The individual or combination treatment with Spirulina platensis and green coffee bean extracts decreased obesity-induced hyperlipidemia and they can be potentially used to treat obesity. Abbreviations: ACC: Acetyl Co-A carboxylase; AF: Activity factor; ALP: Alkaline phosphatase; ALT: Alanine amino transferase; AMP: Adenosine monophosphate; AMPK: AMP activated protein kinase; AST: Aspartate amino transferase; BMI: Body mass index; cAMP: Cyclic adenosine monophosphate; CBE: Coffee bean extract; CVD: Cardiovascular disorder; DRD4: Dopamine receptor D4; FAS: Fatty acid synthase; FFA: Free fatty acid; FTO: Fat mass and obesity gene; GOD: Glucose oxidase; G-6-P: Glucose-6-phosphatase; HDL-C: High-density lipoprotein-cholesterol; HFD: High-fat diet; HMGRCR: 3-hydroxy-3-methylglutaryl CoA reductase; LDL-C: Low-density lipoprotein-cholesterol; LXR α: Liver X receptor α; MC4R: Melanocortin 4 receptor gene’; NO: Nitric oxide; PGC-1α: Peroxisome proliferator-activated receptor gamma coactivator-1 alpha; PPARα: Peroxisome proliferator-activated alpha; PPARγ: Peroxisome proliferator-activated receptor γ; REE: Resting energy expenditure; ROS: Reactive oxygen species; TC: Total cholesterol; T2DM: Type 2 diabetes; TG: Triacyl glycerol; TP: Total protein; VLDL: Very low-density lipoprotein; WLS: Weight loss surgery

PSV-26 The influence of breed, growing phase nutrition, and finishing system (pasture vs. feedlot) on the relative abundance of mRNA associated with lipid metabolism in longissimus muscle

Abstract The objective of this experiment was to evaluate the influence of beef cattle breed, nutritional treatment, and finishing strategy (pasture vs. feedlot) on the relative abundance of mRNA associated with lipid metabolism in longissimus muscle (LM). Eighty-three uncastrated males from three genetic groups: Nellore (N), ½ Angus x ½ Nellore (A) and ½ Senepol x ½ Nellore (S), were randomly assigned to dietary treatments supplied during the growing phase. Treatments consisted of mineral supplement or concentrate supplement (0.3% BW). Following the growing phase, two replicates within each treatment x breed group were randomly assigned to one of two finishing systems (Figure 1): Pasture grazing + concentrate (PC) and Conventional feedlot (CF), both are supplemented at 2% BW/had/d (Table 1). After 209 days the animals were slaughtered, an LM sample (5.0g) was removed between 12th and 13th ribs from each animal and frozen (liquid nitrogen). The relative abundance of mRNA associated with lipid metabolism was measured by qRT-PCR. Target genes include PPARα, PPARγ, SREBP1c, SCD1, LPL, FBP4, CPT2, ACOX and ACCα. The ∆∆ Ct was used to calculate the data and analyzed using the mixed procedures of SAS for a 3×2×2 factorial arrangement of treatments. Mineral supplementation during the growing phase increased (P &amp;lt; 0.05) the relative abundance of mRNA of PPARα and PPARγ compared to concentrate. The relative abundance of PPARγ, SREBP1c, SCD1, FABP4, ACOX, was greater (P &amp;lt; 0.05) in CF compared to PC. Angus LM muscle showed greater (P &amp;lt; 0.05) relative abundance of SREPB1c compared to Nellore and Senepol. The muscle of Nellore and Angus had greater (P &amp;lt; 0.05) relative abundance of PPARα SCD1 and lower (P &amp;lt; 0.05) of PPARγ when compared to Senepol breed. Therefore, in this study, the conventional feedlot and Angus are more positively associated with differential expression of adipogenic genes.

Peroxisome Proliferator-Activated Receptors (PPARs) levels in spermatozoa of normozoospermic and asthenozoospermic men

ABSTRACTInterest in the role of male factor in infertility continues to mount with defects related to sperm movement considered as one of the more severe forms of subfertility. The peroxisome proliferator-activated receptor gamma (PPARγ) primarily regulates the expression of target genes involved in energy control as well as lipid and glucose metabolism. Although the pivotal roles of these receptors on female fertility have been reported, there are limited studies addressing PPARs role(s) in the male. This study was designed to determine and compare PPARα, PPARβ and PPARγ mRNA expression in sperm cells of normozoospermic and asthenozoospermic men. In addition, flow cytometric analyses, immunofluorescence and western blot were used to evaluate PPARγ protein levels in spermatozoa. We have compared the sperm PPARs mRNA relative expression in 27 normozoospermic and 28 asthenozoospermic samples and monitored sperm PPARγ protein levels in 39 normozoospermic and 40 asthenozoospermic samples using flow cytometry. We have also assessed in a sub-group of seven normozoospermic and eight asthenozoospermic samples, PPARγ protein levels by western blotting. Relative expression of PPARγ mRNA in normozoospermic men was found to be significantly higher (P = 0.004) than in asthenozoospermic men while PPARα and PPARβ relative expression was similar in the two groups. Likewise, PPARγ showed a positive correlation with motility (r = 0.34; P < 0.05), sperm concentration (r = 0.33) and the percentage of progressive motile spermatozoa (r = 0.31). In agreement with the mRNA behavior, sperm PPARγ protein levels as measured by flow cytometry (P = 0.066) and western blot (P = 0.089) showed a tendency to be higher in normozoospermic than asthenozoospermic men. The present study proposes a link between PPARγ gene expression level and motility in human sperm.Abbreviations: PPARs: Peroxisome Proliferator-Activated Receptors; CASA: Computer Assisted Semen Analysis; TFA: Trans Fatty Acids; HTF: Human Tubal Fluid; PBS: Phosphate-Buffered Saline; PPP: Pentose Phosphate Pathway; PI3K: Phosphoinositide 3-Kinase; G6PDH: Glucose 6-Phosphate Dehydrogenase

Berberine Induces CYP2J2 Expression in Human U251 Glioma Cells via Regulation of Peroxisome Proliferator-Activated Receptor Alpha

Background: Berberine is a promising natural drug that has a potential therapeutic effect on neurodegenerative diseases. Objectives: Using U251 cells in vitro, we investigated whether berberine exerts its neuroprotective effect via regulation of CYP2J2. Method: After pretreatment with increasing concentrations (1, 3, and 10 μmol/L) of berberine for 0.5 h, U251 cells were stimulated with 1 μg/mL of lipopolysaccharide (LPS). Cell viability was measured 24 h later using a CCK8 kit. mRNA and protein levels of CYP2J2 and peroxisome proliferator-activated receptor alpha (PPARα) were measured by quantitative real-time-polymerase chain reaction and western blotting, respectively, after 24 h of exposure to 1, 3, or 10 μmol/L berberine. Fluorescence immunocytochemistry was also used to evaluate PPARα protein expression after treatment of U251 cells with 10-μmol/L berberine for 24 h. Transient transfection (cotransfection with the plasmid of PPARα- and RXRα-containing) followed by luciferase and chromatin immunoprecipitation assays was used to elucidate the molecular mechanism underlying the observed effects. Results: Compared to the control, LPS-induced U251 cell death was attenuated by berberine in a dose-dependent manner. After 24 h, cell viability was found to be 52.3% (p < 0.05), 66.2% (p < 0.01), and 70.9% (p < 0.001) using 1, 3, and 10 μmol/L berberine treatment, respectively. At these concentrations, berberine increased the CYP2J2 mRNA levels by 1.31-fold (p < 0.05), 1.48-fold (p < 0.01), and 1.88-fold (p < 0.01), respectively, and increased the PPARα mRNA levels 1.17-fold (p < 0.05), 1.29-fold (p < 0.05), and 1.53-fold (p < 0.01), respectively, compared with the respective control groups. In addition, the CYP2J2 and PPARα protein level was also significantly upregulated in U251 cells by berberine (concentrations in 1, 3, and 10 μmol/L) in a dose-dependent manner, compared with the respective control groups. Further investigation indicated that berberine enhances the heterodimerization of PPARα and RXRα, which together bind to the CYP2J2 promoter to induce the expression of CYP2J2 in U251 cells. Conclusion: Upon exposure of U251 cells to berberine, CYP2J2 expression is induced as a result of PPARα stimulation, resulting in a neuroprotective effect.

Maresin 1 Regulates Hepatic FGF21 in Diet-Induced Obese Mice and in Cultured Hepatocytes.

To study the effects of Maresin 1 (MaR1), a docosahexaenoic-acid-derived lipid mediator, on fibroblast growth factor 21 (FGF21) production and to characterize the tissue-specific regulation of Fgf21 and its signaling pathway in liver, skeletal muscle, and white adipose tissue (WAT). Diet-induced obese (DIO) mice are treated with MaR1 (50µg kg-1 , 10 days, oral gavage) and serum FGF21 levels and liver, muscle and WAT Fgf21, β-Klotho, Fgfr1, Egr1, and cFos mRNA expression are evaluated. Additionally, MaR1 effects are tested in mouse primary hepatocytes, HepG2 human hepatocytes, C2C12 myotubes, and 3T3-L1 adipocytes. In DIO mice, MaR1 decreases circulating FGF21 levels and HFD-induced hepatic Fgf21 mRNA expression. MaR1 increases hepatic β-Klotho, Egr1, and cFos in DIO mice. In WAT, MaR1 counteracts the HFD-induced downregulation of Fgf21, Fgfr1, and β-Klotho. In muscle, MaR1 does not modify Fgf21 but promoted Fgfr1 expression. In mouse primary hepatocytes, MaR1 decreases Fgf21 expression and downregulated Pparα mRNA levels. In HepG2 cells, MaR1 reverses the increased production of FGF21 and the downregulation of FGFR1, Β-KLOTHO, EGR1, and cFOS induced by palmitate. Preincubation with a PPARα antagonist prevents MaR1 effects on FGF21 secretion. The ability of MaR1 to modulate FGF21 can contribute to its beneficial metabolic effects.

Fiber and phenolic compounds contribution to the hepatoprotective effects of mango diets in rats fed high cholesterol/sodium cholate.

The present study evaluated the contribution of mango fiber (MF) and mango phenolic compounds (MP) to the hepatoprotective effect of freeze-dried mango pulp (FDM) cultivar (cv.) "Ataulfo" diets in high cholesterol/sodium cholate (HCC)-fed rats. Male Wistar rats were fed with a HCC diet for 12 weeks, either untreated, or supplemented with MF, MP, FDM, or a control diet (no HCC; n = 6/group). All mango treatments significantly decreased hepatic cholesterol deposition and altered its fatty acid profile, whereas MF and MP mitigated adipose tissue hypertrophy. MF caused a lower level of proinflammatory cytokines (IL-1α/β, IFN-γ, TNF-α) whereas FDM increased the anti-inflammatory ones (IL-4, 6, 10). Mango treatments increased catalase (CAT) activity and its mRNA expression; superoxide dismutase (SOD) activity was normalized by MF and FDM, but its activity was unrelated to its hepatic mRNA expression. Changes in CAT and SOD mRNA expression were unrelated to altered Nrf2 mRNA expression. Higher hepatic PPARα and LXRα mRNA levels were found in MP and MF. We concluded that MF and MP are highly bioactive, according to the documented hepatoprotection in HCC-fed rats; their mechanism of action appears to be related to modulating cholesterol and fatty acid metabolism as well as to stimulating the endogenous antioxidant system.

Toll-like receptors control p38 and JNK MAPK signaling pathways in rat astrocytes differently, when cultured in normal or high glucose concentrations

Astrocytes play a vital role in regulating central nervous system inflammation, energy metabolism and brain homeostasis. Unlike macrophages and microglia, which are cells of myeloid ancestry, astrocytes are of ectodermal origin. However, regulatory specificities of signaling pathways connecting inflammatory and metabolic processes are still largely unknown. We analyzed firstly cellular responses to toll-like receptor (TLR) agonists and secondly, modulation of the mRNA of the three isoforms of the transcription factors PPARs (peroxisome proliferator-activated receptors) in primary rat astrocytes exposed to normal glucose (5.5 mM) and high glucose (25 mM). Cell culturing of rat brain astrocytes for 2 days in high glucose did not alter cellular morphology, but i) enhanced the release of TNFα that was induced by TLR4 agonist LPS or TLR3 agonist PIC and the synthesis of prostaglandin E2 (PGE2), ii) changed the signaling pathways of TLR4/MAPK (increase in p38 MAPK, and decrease in JNK activities at early stages of TLR activation) and iii) modulated mRNA expression of PPARs. High glucose cultivation reduced PPARα and PPARβ mRNA levels, without altering PPARγ mRNA level and changed the sensitivity of expressions to agonists of TLR1/2 (PGN), TLR4 (LPS), TLR3 (PIC), and TLR5 (FGN). Differences between low and high glucose-adapted cells were obtained for agonists of TLR1/2 (PPARα, PPARβ), TLR4 (PPAR β), TLR3 (PPARα). In the TLR4/p38/PPARβ signaling pathway, there was a stimulatory connection in normal glucose but an inhibitory connection in high glucose. TLR4/JNK/activated PPARβ, TLR4/JNK/inhibited PPARγ both in cells adapted to normal or high glucose, but PPARα expression was not affected. As PPARs in astrocytes are involved in inflammatory processes in the form of the recently published PPAR triad, the changes in expression revealed here are most likely resulting in implications of high glucose in inflammatory processes. Our data underline the complexity of multiple regulatory interactions between inflammatory responses and energy metabolism in astrocytes.

Hydrogen sulfide regulates circadian-clock genes in C2C12 myotubes and the muscle of high-fat-diet-fed mice

Hydrogen sulfide (H2S) is an endogenous novel gasotransmitter which is implicated in the pathophysiology of the metabolic syndrome. Core clock genes (CCG) and its controlled genes disruption is implicated in the progression of metabolic syndrome. We examined whether H2S has any effect on CCG in the skeletal muscle of mice fed a high-fat diet (HFD) and in myotubes. In the muscle of HFD-mice, the expression of H2S biosynthesis enzyme genes (CSE, CBS, and 3-Mpst) along with antioxidant genes (GCLC, GCLM, GSS, and GSR) involved in GSH biosynthesis and recycling were reduced significantly, but the oxidative stress (OS) increased. Expression of the CCG (Bmal1, Clock, RORα, Cry2, Per2) and clock-controlled genes (PPARγ, PGC-1α, RXRα) was downregulated, whereas the levels of PPARα mRNA were upregulated. Similar to that in the muscle of HFD-mice, in vitro myotubes exposed to high glucose or palmitate to mimic metabolic syndrome, showed an increased OS and decreased in CSE mRNA, H2S production and CCG mRNA levels were also downregulated. TNF and MCP-1 treatment on the myotubes was similar to that observed in HFD-muscle, with that the Rev-erbα mRNA was upregulated. Inhibition (siRNA/pharmacological inhibitors) of both CSE and GCLC (the rate-limiting enzyme in GSH biosynthesis) decreased H2S, and increased OS; Bmal1 and Clock mRNA levels were downregulated, while Rev-erbα increased significantly in these conditions. CSE KD myotubes were post-treated with an H2S donor partially restored the mRNA levels of core clock genes. These findings report that the deficiencies of H2S/GSH impair expression of CCG and treatment with H2S donor or GSH precursor exert a positive effect over CCG. Thus, suggest that H2S as a new endogenous factor for regulating circadian clock, and its donors could provide a novel chrono-pharmacological therapy to manage metabolic disorders.

Effects of Paramylon Extracted from Euglena gracilis EOD-1 on Parameters Related to Metabolic Syndrome in Diet-Induced Obese Mice.

Paramylon (PM), a type of β-glucan, functions like dietary fiber, which has been suggested to exert a protective effect against obesity. We evaluated the potential beneficial effects of PM powder on obesity in mice. Male C57BL/6J mice were fed a high-fat diet supplemented with either 2.5 or 5% PM powder, extracted from Euglena gracilis, for 74 days. Growth parameters, abdominal fat content, serum biochemical markers, hepatic lipid accumulation and hepatic mRNA expression were measured. Dietary supplementation with PM resulted in decreased food efficiency ratios and abdominal fat accumulation. Dose-dependent decreases were observed in postprandial glucose levels, serum low-density lipoprotein (LDL)-cholesterol, and serum secretary immunoglobulin A (sIgA) concentrations. PM supplementation increased peroxisome proliferator-activated receptor α (PPARα) mRNA expression in the liver which is suggested to induce β-oxidation through activation of acyl-coenzyme A oxidase (ACOX), carnitine palmitoyltransferase (CPT) and fatty acid transport protein 2 (FATP2) mRNA expression. Changes in fatty acid metabolism may improve lipid and glucose metabolism. In conclusion, a preventive effect against obesity was observed in mice given a PM-enriched diet. The mechanism is suggested to involve a reduction in both serum LDL-cholesterol levels and the accumulation of abdominal fat, in addition to an improvement in postprandial glucose concentration.

Gene expressions of de novo hepatic lipogenesis in feline hepatic lipidosis.

ObjectivesThe aim of this study was to evaluate if de novo hepatic lipid synthesis contributes to fatty acid overload in the liver of cats with feline hepatic lipidosis (FHL).MethodsLipogenic gene expression of peroxisome proliferator-activated receptor-alpha (PPAR-α), peroxisome proliferator-activated receptor-gamma (PPAR-γ), fatty acid synthase (FASN) and sterol regulatory element-binding factor (SREBF1) were evaluated using quantitative RT-PCR in liver tissue of six cats with FHL and compared with the liver tissue of eight healthy cats.ResultsIn liver tissue, PPAR-α, PPAR-γ and FASN mRNA expression levels were not significantly different (P >0.12, P >0.89 and P >0.5, respectively) in the FHL group compared with the control group. SREBF1 gene expression was downregulated around 10-fold in the FHL group vs the control group (P = 0.039).Conclusions and relevanceThe downregulation of SREBF1 in the liver tissue of cats with FHL does not support the hypothesis that de novo lipogenesis in the liver is an important pathway of fatty acid accumulation in FHL.

Effects of lysine deficiency or excess on growth and the expression of lipid metabolism genes in slow-growing broilers

This experiment was conducted to evaluate the effects of lysine deficiency or excess on growth and the expression of lipid metabolism genes in slow-growing birds. A total of 360 one-day-old chicks were randomly divided into 3 groups, with 6 replicates of 20 birds each. The birds fed the basal diet with a total lysine 0.60% (LL), 1.00% (ML), or 1.40% (HL). The amount of lysine (ML) as the control group, LL and HL as the experimental group, the trial period last 3 wk. The results showed that compared with ML, LL significantly decreased average daily gain and average daily feed intake and remarkably increased feed conversion ratio of birds at 21 day old (P < 0.01), while the above indices in HL had no significant effects (P > 0.05). Besides, LL reduced the pectoral muscle rate (P < 0.01) and decreased the percentage of abdominal fat significantly (P < 0.05). In addition, compared with ML, the expression of fatty acid binding protein 1 (FABP1), acetyl-CoA carboxylase (ACC), malic enzyme (ME), and sterol regulatory element binding protein 1 (SREBP1c) mRNA of liver in LL was significantly decreased (P < 0.05), and the expression of cholesteryl ester transfer protein (CETP) mRNA was significantly increased (P < 0.01), whereas LL had no significant effects on the expression of peroxisome proliferator activated receptor alpha (PPARα) mRNA (P > 0.05). Moreover, compared with ML, HL significantly reduced the expression of FABP1, ACC, ME, SREBP-1c, and PPARα mRNA in the liver (P < 0.05), and had no significant effects on the expression of CETP mRNA (P > 0.05). The results of current research suggest that dietary lysine deficiency could reduce the growth and fat deposition of slow-growing broilers mainly by downregulating the expression of lipid synthesis genes.