PPARgamma Gene Research Articles

Effects of Switching from Stavudine to Raltegravir on Subcutaneous Adipose Tissue in HIV-Infected Patients with HIV/HAART-Associated Lipodystrophy Syndrome (HALS). A Clinical and Molecular Study

HIV-1/HAART-associated lipodystrophy syndrome (HALS) has been associated with exposure to stavudine (d4T) through mitochondrial dysfunction. We performed a 48-week study to assess the effects of switching from d4T to raltegravir (RAL) on metabolic and fat molecular parameters of patients with HALS. Forty-two patients with HALS and a median exposure to d4T > 7 years were switched to RAL and followed for 48 weeks. Fasting metabolic tests, HIV RNA, CD4 cell count, and fat measured by DEXA were obtained at baseline and week 48. mtDNA and gene transcripts for PPAR gamma, adiponectin, cytochrome b, Cox IV, TNF alpha, MCP-1 and CD68 were assessed in paired subcutaneous fat tissue biopsies. Lipid parameters, fasting glucose, insulin, and HOMA-IR did not change significantly. Whole body fat (P = 0.0027) and limb fat mass (P<0.0001) increased from baseline. Trunk/limb fat ratio (P = 0.0022), fat mass ratio (P = 0.0020), fat mass index (P = 0.0011) and percent leg fat normalized to BMI (P<0.0001) improved after 48 weeks. Relative abundance of mtDNA, expression of PPAR gamma, adiponectin, Cyt b, and MCP-1 genes increased, whereas Cox IV, TNF alpha, and CD68 did not change significantly from baseline. Switching from d4T to RAL in patients with HALS is associated with an increase in limb fat mass and an improvement in markers of adipocyte differentiation and mitochondrial function in SAT.

Exchange protein activated by cyclic AMP is involved in the regulation of adipogenic genes during 3T3-L1 fibroblasts differentiation

Adipogenesis is stimulated in 3T3-L1 fibroblasts by a combination of insulin, dexamethasone and isobutylmethylxanthine, IBMX, (I+D+M). Two transcription factors are important for the acquisition of the adipocyte phenotype, C/EBP beta (CCAT enhancer-binding protein beta) and PPAR gamma (peroxisome proliferator-activated receptor gamma). IBMX increases cAMP content, which can activate protein kinase A (PKA) and/or EPAC (exchange protein activated by cAMP). To investigate the importance of IBMX in the differentiation mixture, we first evaluated the effect of the addition of IBMX on the increase of C/EBP beta and PPAR gamma and found an enhancement of the amount of both proteins. IBMX addition (I+D+M) or its replacement with a cAMP analogue, dibutyryl-cAMP or 8-(4-chlorophenylthio)-2-O'-methyl-cAMP (8CPT-2-Me-cAMP), the latter activates EPAC and not PKA, remarkably increased PPAR gamma mRNA. However, neither I+D nor any of the inducers alone, increased PPAR gamma mRNA to a similar extent, suggesting the importance of the presence of both IBMX and I+D. It was also found that the addition of IBMX or 8CPT-2-Me-cAMP was able to increase the content of C/EBP beta with respect to I+D. In agreement with these findings, a microarray analysis showed that the presence of either 8CPT-2-Me-cAMP or IBMX in the differentiation mixture was able to upregulate PPAR gamma and PPAR gamma-activated genes as well as other genes involved in lipid metabolism. Our results prove the involvement of IBMX-cAMP-EPAC in the regulation of adipogenic genes during differentiation of 3T3-L1 fibroblasts and therfore contributes to elucidate the role of cyclic AMP in this process.

Peroxisome proliferator activated receptor γ2 gene Pro12Ala gene polymorphism in type 2 diabetes and its relationship with diabetic nephropathy.

Despite intensive research, a genetic background of type 2 diabetes is still unknown as are causes of differences in the clinical course of the disease. This supports the hypothesis for factors of genetic susceptibility (or protection) to diabetic nephropathy. This study aimed to examine the association between Pro12Ala polymorphism in the PPARgamma2 gene with both type 2 diabetes and the development of diabetic nephropathy. The study included seventy subjects classified into three groups: healthy control group (20), diabetics without nephropathy (25), and diabetics with nephropathy (25). All members of the study were subjected to the following laboratory investigations: fasting glucose and insulin with calculation of insulin resistance by HOMA IR, HbA1c, urea, creatinine and calculation of creatinine clearance, lipid profile, urinary protein and/or microalbumin. Determination of Pro12Ala polymorphism in PPARgamma2 gene was done by PCR/RFLP. As regards PPARgamma2 Pro12Ala gene polymorphism, the Pro/Pro genotype was the most common in diabetic patients as well as in controls followed by Pro/Ala genotype and Ala/Ala genotypes was the least common one. The genotype distribution of the PPARgamma2 polymorphism was significantly different in diabetic patients compared to controls and in diabetics with nephropathy compared to those without nephropathy (p < 0.05). The allelic frequency of proline (Pro) was significantly higher in diabetic patients than controls (p < 0.001) and also significantly higher in diabetics with nephropathy than without nephropathy (p < 0.05). The odds ratio (OR) of having diabetes for Pro allele carriers was 8 compared with noncarriers (95% CI: 2.66 - 23.98). Our results support the association between the Pro allele of PPARgamma2 gene and both development of type 2 diabetes and diabetic nephropathy.

The Relationship of the Peroxisome Proliferator Activated Receptor-Gamma 2 Exon 2 and Exon 6 Gene Polymorphism in Type 2 Diabetic Patients with and without Diabetic Foot Ulcers

Abstract We aims investigate Turkish type 2 diabetic patients with/without diabetic foot ulcers and healthy group and examined the contribution of the Pro12Ala exon 2 and C478T exon 6 of the PPARgamma gene polymorphisms to the development of diabetic foot ulcers. The PPARgamma genotypes were determined prospectively in 50 patients with diabetic foot ulcers and 50 without diabetic foot ulcers and a control group of 50 healthy individuals. Genotyping of the Pro12Ala exon 2 and C478T exon 6 of the PPAR-gamma gene polymorphisms for all individuals was performed by PCR-RFLP method. The genotype exon 2 and exon 6 distribution did not differ between the control group and the type 2 diabetes mellitus patients (with and without diabetic foot) (P>0.05). The frequency of the polymorphic G allele in exon 2 was no similar for the groups (6% and 1%, respectively) (p 0.05). The evaluation of exon 2 and exon 6, genotype and haplotype did not show statistically significant difference between the patient diabetic foot and without diabetic foot (p>0.05). The Pro12Ala exon 2 and C478T exon 6 of the PPARgamma gene polymorphisms are not an independent risk factor for diabetic foot in Turkish type 2 diabetes mellitus patients. Genetic factors in the pathogenesis of diabetic foot may also show any changes in different populations. Key words: Peroxisome Proliferator Activated Receptor Gene Polymorphism, Type 2 Diabetic Patients, diabetic foot ulcers

The orphan nuclear receptor Ear-2 (Nr2f6) is a novel negative regulator of T cell development

We describe a novel role for the orphan nuclear receptor Ear-2 in regulating T cell development. Retrovirus-mediated overexpression of Ear-2 (EAR-2++) in a bone marrow (BM) transplantation assay resulted in limited T cell development and a greater than tenfold decrease in thymus size and cellularity relative to controls. Ear-2-transduced murine BM hematopoietic stem cells (HSCs) in OP9-DL1 cultures showed a proliferation deficit during days 1-5 after induction of differentiation, which corresponded to increased expression of the cell cycle regulators p21 (cdkn1a) and p27 (cdkn1b), as well as increased expression of Hes1, Notch3, Egr1, and Scl (Tal1) and decreased expression of Gli1, Gfi-1, HoxA9, PU.1, Nrarp, and Tcf1. In addition, there was a block in differentiation at the DN4 to double-positive (DP) transition accompanied by an increase in apoptosis, similar to the deficit seen in the RORγt null mouse. Gene expression profiling revealed that, like the RORγt-deficient mouse, EAR-2++ DP cells had decreased expression of BclXL and increased expression of the proapoptosis gene Bad. In addition, EAR-2++ DP cells had decreased expression of Bcl11b, PU.1, and HoxA9, and increased expression of Id2. Based on these findings, we conclude that EAR-2++ cells were able to migrate to, but not fully repopulate, the thymus because of a cell-intrinsic defect in the proliferation of DN1 cells followed by a block in differentiation from the DN4 to DP stage of T cell development. We conclude that Ear-2 is a novel negative regulator of T-cell development and that downregulation of Ear-2 is indispensable for the proliferation of DN1 cells and the survival of DN4-DP cells.

Influence of the body mass and visceral adiposity on glucose metabolism in obese women with Pro12Pro genotype in PPARgamma2 gene.

Glucose metabolism may be altered in obesity and genotype for PPAR 2 can influence this variable. To evaluate the influence of body mass (BM) and visceral adiposity (VA) in glucose metabolism in morbid obese women with Pro12Pro genotype. Were selected 25 morbidly obese women. Groups were formed according to body mass index (BMI) [G1: 40-45 kg/m² (n = 17); G2: > 45 kg/m² (n = 8)]. Anthropometric, glycemia and insulinemia assessments (fasting, 60 and 120 minutes after high polyunsaturated fatty acids meal) were carried out. The insulin resistance (IR) and insulin sensitivity (IS) were assessed by HOMA-IR and QUICKI respectively. G2 had higher BMI and waist circumference, compared to G1, impaired fasting glucose, low IS and higher IR. The postprandial glucose was normal, but there was a higher insulin peak one hour after the meal in G2. Increased BM and VA were associated with worse glucose metabolism suggesting metabolic differences between morbid obese with Pro12Pro genotype.

Association of Genetic Polymorphism of Peroxisome Proliferator‐Activated Receptor‐Gamma Gene and Polycystic Ovary Syndrome

BackgroundPeroxisome proliferative‐ activated receptor gamma (PPARs) is a nuclear receptor which controls transcription of genes involved in free fatty acid uptake and lipogenesis, and is essential for differention and survival of adipocytes.AimTo investigate the association of Pro12Ala polymorphism in PPAR gamma gene with polycystic ovary syndrome (PCOS) and its related metabolic disorders.Material and MethodThe study consisted of 50 women diagnosed with PCOS and 96 healthy women as a control group. Individuals were genotyped for Pro12Ala genetic polymorphism of PPAR gamma by using polymerase chain reaction‐ restriction fragment length polymorphism. Statistical analysis was done by SPSS program.ResultsFrequency of Pro12/Pro12 and Pro12/Ala12 genotypes were 86% and 14% in PCOS group compared to 77.1% and 22.9% in control group. The carriers of Pro12/Pro12 genotype exhibited a significant greater risk for PCOS compared to carriers of Pro12/Ala12 genotype (OR= 1.8, 95%CI= 0.7–4.6). Furthermore they were at higher risk of different metabolic disorders such as obesity, insulin resistance and diabetes mellitus.ConclusionPro12/Pro12 genotype is a risk factor for PCOS and its related metabolic disorders.Source of Fund: Dubai Medical College for girls.

DNA methylation patterns of peroxisome proliferator‐activated receptor gamma gene associated with liver fibrosis and inflammation in chronic hepatitis B

Peroxisome proliferator-activated receptor gamma (PPAR gamma) is a nuclear receptor that regulates gene expression of inflammatory mediators in liver injury. Hepatitis B virus (HBV) suppresses the PPAR gamma-mediated transactivation in liver cancerous cell lines. However, the role of PPAR gamma in patients with chronic HBV infection has not fully demonstrated. Our present study was firstly to determine the clinical relevance of peripheral PPAR gamma mRNA levels in chronic hepatitis B (CHB) patients, and then, the DNA methylation of PPAR gamma promoter was investigated. Peripheral blood mononuclear cells (PBMCs) were isolated from 91 CHB patients and 18 healthy controls. The mRNA level of PPAR gamma was determined by quantitative real-time PCR; meanwhile, the CpG island methylation was assessed by methylation-specific PCR. CHB patients showed significantly lower mRNA level of PPAR gamma than healthy controls (P=0.005). The mRNA level was decreased in HBV-DNA-positive group than HBV-DNA-negative group (P=0.041). Interaction analysis demonstrated that the DNA methylation pattern was responsible for the suppression of peripheral PPAR gamma transcription in CHB patients (P=0.003). Furthermore, the hypermethylation of PPAR gamma gene promoter was significantly associated with liver inflammation and fibrosis in CHB. In conclusion, DNA methylation patterns were responsible for the decreased mRNA level of peripheral PPAR gamma in CHB patients. Liver inflammation and fibrosis were found to be associated with hypermethylation of PPAR gamma promoter.

Association between PRO12ALA polymorphism of the PPAR-γ2 gene and type 2 diabetes mellitus in Iranian patients

BACKGROUND:Peroxisome proliferator-activated receptor (PPARs) have been identified as ligand-activated transcription factors that belong to the nuclear receptor superfamily. It has been shown that an association exists between Proline 12 alanine (Pro12Ala) polymorphism of PPAR-GAMMA2 (PPAR-γ2) gene and increased risk of type 2 diabetes mellitus (T2DM) in different populations. Therefore, the present study was designed to investigate the association between Pro12Ala polymorphism of PPAR-γ2 gene and T2DM in an Iranian population.MATERIALS AND METHODS:Two hundred unrelated people, including 100 healthy controls and 100 diabetic patients were recruited diagnosed based on American Diabetes Association criteria. Blood samples were used for isolation of genomic deoxyribonucleic acid (DNA). Having extracted the genomic DNA from human blood leukocytes by means of High Pure polymerase chain reaction (PCR) Template preparation kit, we carried out polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) on each blood sample. Then, Genomic DNA was digested by BstU-I restriction enzyme. Thereafter, restriction products were analyzed by means of Polyacrylamide gel electrophoresis and stained by Ethidium Bromide.RESULTS:We found that the frequency of Ala allele in healthy subjects was significantly higher than in diabetic subjects (P = 0003). Moreover, the genotype frequency of Ala/Ala in healthy subjects was significantly higher than in diabetic subjects (P < 0.001). However, the genotype frequency of Ala/Pro in diabetic subjects was significantly higher than in healthy subjects (P < 0.001).CONCLUSION:The present study suggests that polymorphism of PPAR-γ2 gene is associated with T2DM. Furthermore, Ala allele is significantly found in non-diabetic individual’s Iranian population.

Structural Features and Transcriptional Activity of Chicken PPARs (α,β, andγ)

While an understanding of lipid metabolism in chickens is critical for a further improvement of food production, there are few studies concerning differences in lipid metabolism mechanisms between chickens and other species at a molecular level. Chickens have three PPAR gene subtypes (α, β, and γ) that function differently from those present in humans and mice. The chicken PPAR-gamma (cPPARγ) gene is shorter than that in humans and lacks a γ2 isoform. Moreover, in serum-free media, cPPARγ shows high transcriptional activity without exogenous ligands. Luciferase reporter assays were used to examine the effect of sera on cPPAR transcriptional activities and showed that adult bovine serum and chicken serum highly activate cPPARα and β functions. Moreover, we found that bezafibrate induces the transactivation function of cPPARβ, but not human PPARδ (human PPARβ ortholog). This ligand selectivity relies on one amino acid residue (chicken: Val419, human: Met444). These results show the possibilities for unique functions of cPPARs on chicken-specific lipid glucose metabolism. As such, a better understanding of the molecular mechanisms of lipid metabolism in chickens could result in higher productivity for the poultry industry.

The role of the expression of PPAR-gamma gene in the adipogenesis in hemangioma evolution

To investigate the role of the expression of PPAR-gamma gene in the adipogenesis in hemangioma evolution. Routine immunohistochemistry staining of Perilipin A, the marker antigen of adipocytes, was performed to observe the adipogenesis in hemangioma. Immunofluorescence staining of PPAR-gamma, the important transcription factor in promoting adipogenesis, was carried out to observe its location in hemangioma tissue, with the co-staining of alpha-SMA and CD31. And RT-PCR was used to examine the expression of PPAR-gamma gene in hemangioma in different stages. In the evolution of hemangioma, the number of adipocytes increased continuously. And the tumor was replaced by fibrofatty tissue finally. PPAR-gamma was located in the nuclei of perivascular cell in hemangioma tissue. The expression of PPAR-gamma gene in hemangioma increased in the evolution of hemangioma, but still was lower than that in normal fat tissue from children. The expression of PPAR-gamma in the perivascular cells suggests that they may contribute to the adipogenesis in hemangioma involution.

Abstract A110: Associations between genetic variations in inflammation and innate immunity pathways and colorectal cancer

Abstract A strong association between chronic inflammation and colorectal cancer (CRC) risk has been observed. We postulated that common variation in genes involved in the inflammation and innate immunity pathways may result in a chronic pro-inflammatory state that may lead to increased CRC risk. We comprehensively examined the associations of 724 tagSNPs in 37 key genes in these pathways with CRC in two large well-characterized studies. The screening phase included 2,322 discordant sibships (2,535 cases, 3,915 unaffected sibling controls) from the Colon Cancer Family Registry. Nineteen gene regions spanned by 99 top hits from main-effect or interaction-effect analyses in phase 1 were followed up by genotyping 351 well-chosen tagSNPs in 4,783 subjects (2,153 cases, 2,630 controls) in five ethnic groups in the Multiethnic Cohort Study. One SNP rs9858822 in PPAR gamma gene was statistically significantly associated with CRC in the replication phase (p=1.5 × 10-4, odds ratio per allele = 1.36), based on a conservative Bonferroni-corrected significance level 0.05 adjusted for the “effective” number of markers (n=306). The risk allele C was common (frequency 0.3) in African Americans but was rare (frequency &amp;lt; 0.05) in Whites, Japanese Americans, Latinos and Native Hawaiians. We did not detect effect modification by race/ethnicity, BMI, or use of aspirin. Strong, although not statistically significant, associations were also observed for SNPs in the neighborhood region of rs9858822 (p-values as low as 2.5 × 10-4). Further replication is needed. Citation Format: Hansong Wang, Darin Taverna, Daniel O. Stram, Iona Cheng, Lynne R. Wilkens, Terrilea Burnett, Polly A. Newcomb, Laurence N. Kolonel, Brian E. Henderson, David Duggan, Cornelia M. Ulrich, Loic Le Marchand. Associations between genetic variations in inflammation and innate immunity pathways and colorectal cancer. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr A110.

Peroxisome proliferator-activated receptor gamma inhibits hepatic fibrosis in rats

Hepatic fibrosis is a necessary step in the development of hepatic cirrhosis. In this study we used lentiviral vector-mediated transfection technology to evaluate the effect of peroxisome proliferator-activated receptor gamma (PPAR-gamma) on rat hepatic fibrosis. Hepatic fibrosis in rats was induced by CCl4 for 2 weeks (early fibrosis) and 8 weeks (sustained fibrosis). The rats were randomly divided into four groups: normal control, fibrosis, blank vector, and PPAR-gamma. They were infected with the recombinant lentiviral expression vector carrying the rat PPAR-gamma gene by portal vein injection. The liver of the rats was examined histologically and hydroxyproline was assessed. In vitro primary hepatic stellate cells (HSCs) were infected with the recombinant lentiviral expression vector carrying the rat PPAR-gamma gene. The status of HSC proliferation was measured by the MTT assay. The protein levels of PPAR-gamma, alpha-smooth muscle actin (alpha-SMA) and type I collagen expression were evaluated by the Western blotting method. In vitro studies revealed that expression of PPAR-gamma inhibited expression of alpha-SMA and type I collagen in activated HSCs (P<0.01) as well as HSC proliferation (P<0.01). In vivo experiments indicated that in the early hepatic fibrosis group, the hydroxyproline content and the level of collagen I protein in the liver in the PPAR-gamma transfected group were not significantly different compared to the hepatic fibrosis group and the blank vector group; whereas the expressions of PPAR-gamma and alpha-SMA were different compared to the hepatic fibrosis group (P<0.01). In the sustained hepatic fibrosis group, there were significant differences in the hydroxyproline content and the expression of PPAR-gamma, alpha-SMA, and type I collagen between each group. PPAR-gamma can inhibit HSC proliferation and hepatic fibrosis, and suppress alpha-SMA and type I collagen expression.

Prevalence of the Pro12Ala missense mutation in the PPARG2 gene in Kuwaiti patients with primary knee osteoarthritis

BACKGROUND AND OBJECTIVES:Peroxisome proliferator–activated receptors (PPARs) play an important role in a number of cellular and metabolic functions. This study was carried out to determine the prevalence of a missense mutation (Pro12Ala) in the PPARG2 gene in Kuwaiti Arab patients with primary knee osteoarthritis (OA) and healthy controls with the aim of identifying a possible association.DESIGN AND SETTING:A prospective cross-sectional study carried out at three major teaching hospitals (referral centers) in the country over a one-year period.PATIENTS AND METHODS:The prevalence of PPARG2 gene Pro12Ala missense mutation was determined in 104 Kuwaiti Arab patients with primary knee OA and 111 ethnically matched healthy controls. The prevalence of this Pro12Ala missense mutation was also determined in clinical subgroups of OA patients divided on the basis of age at onset, function and radiologic grading.RESULTSThe Pro-Pro genotype of the PPARG2 gene Pro12Ala missense mutation was detected in 95/104 (91.3%) cases compared to 111/111 (100%) in the control subjects. The heterozygous Pro-Ala genotype was detected in 9/104 (8.7%) of the OA patients, while it was not detected in any of the controls. The Ala-Ala genotype was not detected in any of the OA patients or the controls. No significant differences were detected in the PPARG2 gene Pro12Ala genotypes in the subgroups of patients classified on the basis of age at onset, functional assessment using Lequesne’s functional index, and radiological grading using Kellgren-Lawrence (K-L) grading.CONCLUSIONSThis study found no significant association between the PPARG2 gene Pro12Ala missense mutation and knee OA. However, the presence of the Pro-Pro genotype of the PPARG2 gene mutation has a protective effect against development of OA.

Glucose regulates fatty acid binding protein interaction with lipids and peroxisome proliferator-activated receptor α

Although the pathophysiology of diabetes is characterized by elevated levels of glucose and long-chain fatty acids (LCFA), nuclear mechanisms linking glucose and LCFA metabolism are poorly understood. As the liver fatty acid binding protein (L-FABP) shuttles LCFA to the nucleus, where L-FABP directly interacts with peroxisome proliferator-activated receptor-α (PPARα), the effect of glucose on these processes was examined. In vitro studies showed that L-FABP strongly bound glucose and glucose-1-phosphate (K(d) = 103 ± 19 nM and K(d) = 20 ± 3 nM, respectively), resulting in altered L-FABP conformation, increased affinity for lipid ligands, and enhanced interaction with PPARα. In living cells, glucose stimulated cellular uptake and nuclear localization of a nonmetabolizable fluorescent fatty acid analog (BODIPY C-16), particularly in the presence of L-FABP. These data suggest for the first time a direct role of glucose in facilitating L-FABP-mediated uptake and distribution of lipidic ligands to the nucleus for regulation of PPARα transcriptional activity.

Study on effect of total saponins from Semen Nigellae on inflammatory mediators and ERK/MAPK pathway in stimulated macrophage

To study the anti-inflammatory mechanism of total saponins from Semen Nigellae (TSSN). IFN-gamma plus LPS stimulated RAW 264. 7 macrophage has been used as inflammatory experimental model. Griess reaction for nitric oxide production, FRAP assay for total antioxidant capacity, RT-PCR for mRNA expression and Western blot for protein expression examination were performed. TSSN inhibited NO production in a dose-dependent manner. The gene and protein expression of iNOS were also suppressed by the herb extract. TSSN treatment significantly attenuated mRNA of inflammatory mediators such as COX-2, IL-1beta, IL-6 while increased PPAR-gamma gene and protein expression. Furthermore, phosphorylation of ERK (p-ERK) was markedly inhibited by TSSN. TSSN suppressed pro-inflammatory mediators such as COX-2, IL-1beta, IL-6 and increased anti-inflammatory mediator PPAR-gamma expression. Meanwhile, TSSN inhibited over production of NO and iNOS expression through ERK/MAPK pathway.

The Relationship between Peroxisome Proliferator-Activated Receptor-γ and Renin: A Human Genetics Study

Peroxisome proliferator-activated receptor gamma (PPARgamma) agonists often cause volume retention and edema. A relationship between PPARgamma and renin may play a role in this process. The aim was to examine the relationship between the PPARgamma gene and plasma renin activity (PRA) levels in human hypertension. A candidate gene association study was conducted with two distinct groups of human participants: Caucasian hypertensives (n = 395) and African-American hypertensives (n = 55). Single nucleotide polymorphisms of the PPAR(Upsilon) gene were analyzed. Phenotype studies were conducted after participants consumed a low-salt diet (10 mmol/d) for 7 d and included PRA and aldosterone measurements before and after a 60-min angiotensin II infusion (3 ng/kg x min). Participants homozygous for the minor allele of rs2959272 (CC) had significantly higher PRA levels at baseline (P = 0.016) than major allele carriers (AA, AC) in Caucasian-hypertensive participants. The association of the C allele carrier status with increased PRA levels was replicated in the group of African-American hypertensive participants (P = 0.027). The Fisher's combined P value for both observations was significant (P = 0.002). These results demonstrate the first known association between a PPARgamma single nucleotide polymorphism and alterations in PRA levels in humans with hypertension. This link between PPARgamma and renin raises the possibility of a genetically based mechanism for the increased volume retention and edema in some users of PPARgamma agonists.

Influence of fat diet in glicidic metabolism in obese women with Pro12Pro genotype in PPARgamma2 gene

The peroxisome proliferator-activated receptor gamma 2 (PPARgamma2) is an adipogenic transcription factor that influences in insulin resistance (IR) in the presence of agonists such as polyunsaturated fatty acids (PUFA). Evaluate the influence of dietary fat in glicidic metabolism in morbidly obese women with Pro12Pro genotype in the gene PPARgamma2. Were selected 25 women with genotype Pro12Pro. The fat intake was estimated by food records, being used for the division of groups, GA (until 30% of the total energy expenditure (TEE)) and GB (greater than 30% of the TEE). Biochemical and anthropometric evaluations were conducted in fasting, following the test meal high in n-6 PUFA and postprandial biochemical evaluations. IR and insulin sensitivity (IS) were assessed by HOMA-IR (Homeostasis Model Assessment) and QUICKI (Quantitative Insulin Sensitivity Check Index), respectively. GA presented normal HOMAIR and QUICKI. GB presented higher body mass index (BMI), HOMA-IR, saturated fatty acids (SFA) and monounsaturated (MUFA) intake higher, compared with GA (p < 0.05). In GA, the MUFA intake was negatively correlated with HOMA-IR, fasting glucose and insulin, and positively with QUICKI. The fat and SFA intake contributed to the increase in body mass and IR. However, MUFA intake may have reduced the impact of high fat diet in glicidic metabolism. It is suggested that obese women with Pro12Pro genotype in the PPARgamma2 gene avoid high fat and SFA diets, prioritizing MUFA for controlling obesity and improving the IS.

DIFFERENTIAL EFFECT OF TELMISARTAN AND AMLODIPINE ON MONOCYTE CHEMOATRACTANT PROTEIN-1 AND PEROXISOME PROLIFERATOR ACTIVATED RECEPTORS GAMMA GENE EXPRESSION IN HYPERTENSIVES: PP.13.482

Objective: Monocyte chemoattractant protein-1 (MCP-1), and peroxisome proliferator-activated receptors gamma (PPAR-gamma) play a significant role in monocytes activation, vascular inflammation and atherogenesis. Angiotensin receptor blockers and calcium blockers are both antihypertensive drugs with established efficacy and favorable safety profile.We investigated the effect of telmisartan – angiotensin II type 1 receptor blocker with PPAR-gamma agonist activity – and amlodipine on activation state of peripheral blood monocytes with respect to MCP-1, and PPAR-gamma mRNA expression in patients with essential hypertension. Design and Method: We recruited 28 previously untreated patients with mild or moderate essential hypertension (16 male, aged 61 ± 10 years) who were randomly assigned to receive treatment with either telmisartan (n = 14) or amlodipine (n = 14), achieving a mean blood pressure below 140/80 mmHg. Blood samples were taken before and 3 months after therapy initiation. Mononuclear cells were isolated using anti-CD14 + antibodies and mRNAs were estimated by real-time quantitative reverse transcription-PCR and expressed as fold induction. Results: There were no significant changes in blood pressure (BP), glucose, and lipid parameters between baseline and 12 weeks after randomization in either group. Both treatments reduced all blood pressure components significantly (p < 0.001). In contrast with amlodipine group, MCP-1 gene expression was significantly downregulated after treatment with telmisartan (fold induction in telmisartan group 0.8 ± 0.35 versus 1.87 ± 0.9 in the amlodipine group, p = 0.03). Additionally, PPAR-gamma gene expression showed a significant increase in telmisartan treated patients (fold induction in telmisartan group 2.8 ± 4.4 versus 5.3 ± 2.9 in the amlodipine group, p = 0.04). Conclusions: Treatment with telmisartan results in a significant induction of PPAR-gamma gene expression and attenuation of MCP-1 gene expression in peripheral monocytes in patients with essential hypertension. Our findings may provide new insights into the benefits and cardiovascular protection of telmisartan in hypertensive patients.

Mesenchymal stem cells from CD34− human umbilical cord blood

Umbilical cord blood (UCB) is well known to be a rich source of stem cells especially for haematopoietic stem cells (HSCs). Recently, mesenchymal stem cells (MSCs) have also been shown to exist in cord blood. Although MSCs have been described by a subset of surface antigens after expansion, little is known about the cell surface phenotype of undifferentiated MSCs. The aim of this study therefore was to clarify whether undifferentiated MSCs are resident among CD34(-) UCB cells. CD34(+) cells were separated from UCB mononuclear cells (MNCs) by magnetic sorting and the CD34(-) cell fractions were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% foetal calf serum (FCS) and basic-fibroblast growth factor. Isolated CD34(+) cells were also cultured in the same medium. Adherent fibroblast-like cells at passage 3-4 were analyzed by fluorescence-activated cell sorting (FACS) for MSC marker expression , and standard adipogenic, osteogenic and chondrogenic assays were used to investigate their differentiation potentials. After 4-5 weeks in culture, the cells from the CD34(-) fraction became confluent with flat and fibroblast-like morphology. These cells were positively stained for the mesenchymal cell markers CD29, CD73 and CD105. In adipogenic differentiation, the cells showed oil red O positive and expressed FABP4, adipsin and proliferation-activated receptor gamma-2 (PPARgamma2 genes) associated with adipogenesis. In osteogenic differentiation, calcium accumulation and osteocalcin were detected. The cells grown in chondrogenic conditions were positively stained for human aggrecan and expressed collagen type II and Sox-9 genes. In contrast, cells from the CD34(+) fraction failed to generate any cells with MSC morphology under the same culture conditions. Our results showed that UCB contained MSCs which are only resident in the CD34(-) fraction. The MSCs could be induced to differentiate into at least three lineage cell types, adipocytes, osteoblasts and chondrocytes.