Inhibition of PPARγ Gene Particle Downregulates the Differentiation of Rabbit Bone Marrow Mesenchymal Stem Cells into Adipocytes

This study aims to evaluate the effect of peroxisome proliferator-activated receptor (PPAR) γ gene inhibition on the adipogenic differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs). Primary BMSCs were isolated from rabbit bone marrow, cultured, and the markers of BMSCs on cell’s surface were analyzed using flow cytometry. The experiment involved five groups, namely, control: untreated BMSCs; model: BMSCs treated with ethanol; empty siRNA: BMSCs treated with ethanol + empty siRNA; PPARγ: BMSCs treated with ethanol + PPARγ siRNA; and PPARγ inhibitor: BMSCs treated with ethanol + T0070907. RT-PCR and Western blotting were used to detect changes in the expression level of PPARγ, PETALA2 (AP2), lipoprotein lipase (LPL), fatty acid transport protein (FATP) 1, and fatty acid transporter (FAT). Adipocyte count and triacylglycerol content of the model and the empty siRNA groups were considerably greater than the control group ( P < 0.01 ). After the inhibition with PPARγ or T0070907, adipocyte count and triacylglycerol content of the PPARγ and T0070907 groups were significantly reduced ( P < 0.01 ), with no statistically significantly difference than the control group ( P > 0.05 ). The expression levels of PPARγ gene and protein in the model and empty siRNA groups were ominously enhanced than the control group ( P < 0.01 ), and after inhibition with PPARγ or T0070907, the PPARγ gene or protein expression level of PPARγ and T0070907 groups significantly reduced ( P < 0.01 ), with no statistically significance difference compared to the control group ( P > 0.05 ). The expression levels of Ap2, LPL, FATP1, and FAT genes in the model and empty siRNA groups were considerably greater compared to the control group ( P < 0.01 ). Inhibition with PPARγ or T0070907 in the PPARγ and T0070907 groups, respectively, lead to significantly reduced expression levels of adipogenic genes ( P < 0.01 ), with no statistically significance difference compared to the control ( P > 0.05 ). Inhibition of PPARγ gene downregulates the differentiation of BMSCs into adipocytes, indicating its putative role in the expression of adipogenic genes.

PPARγ gene expression analysis in psoriasis treatment

Introduction. PPARγ is the most studied PPAR subtype and is expressed predominantly in adipose tissue, heart, colon, kidney, spleen, intestine, skeletal muscle, liver, macrophages, and skin. In the skin, PPARγ controls the genetic regulation of gene network expression involved in cell proliferation, differentiation, and inflammatory responses. PPARγ (Peroxisome proliferator-activated receptor gamma) has only recently come to be considered a key player in the development and pathogenesis of psoriasis and psoriatic inflammatory conditions.Aim of the study. To study PPARγ gene expression in the affected skin of psoriasis patients in comparison with visually unaffected skin. To study changes in PPARγ gene expression level in psoriasis affected skin in comparison with unaffected skin in patients before and after treatment with low-level laser radiation with a wavelength of 1.27 μm.Materials and methods. Twelve patients with psoriasis participated in the study. Biopsies from unaffected skin areas were taken at a distance of about 3 cm from the affected skin. Analysis was performed by real-time PCR.Results and Discussion. We quantitatively measured PPARγ gene expression using RT-PCR in the affected skin of patients with psoriasis in comparison with visually unaffected skin in the same patients before and after treatment with low-level laser radiation with a wavelength of 1.27 μm (the short-wave part of the infrared range). The study experimentally showed a 1.3 ± 0.27-fold decrease in PPARγ gene expression in the affected skin of psoriasis patients on average. Significant increase in over-expression of PPARγ gene up to 2,13 ± 0,47 times was observed after treatment of patients with low-level laser radiation.Conclusions. PPARγ gene expression may be an indicator of the efficacy of psoriasis treatment at the molecular level, as well as become a new therapeutic target.

Challenges in Managing Metabolic Complications in a Patient With Familial Partial Lipodystrophy Type 3

Familial partial lipodystrophy (FPL) is a rare group of autosomal dominant genetic disorders which causes variable loss of subcutaneous fat from abdomen, thorax or extremities in addition to the numerous metabolic complications like insulin resistance, diabetes mellitus and dyslipidemia1. FPL type 3 was first characterized by Agarwal et al. in 20021, in which peroxisome proliferator-activated receptor-γ (PPARγ) gene was the molecular basis of this disorder. It is extremely rare and so far only 30 patients or so have been recognized with this mutation2. FPL3 is unique because it generally spares the loss of fat from trunk, face and neck region and also presents with more severe metabolic derangements. We report a case of a young female with PPARγ mutation leading to numerous metabolic complications. A 19 year old female with FPL3 was seen by adult endocrinology as a transition from pediatric endocrinology. She was found to have hypertriglyceridemia on routine labs done at the age of 11. Patient reported loss of subcutaneous fat from her extremities and eruptive xanthoma on flexor surfaces at the time of diagnosis along with a positive family history of hypertriglyceridemia induced pancreatitis and Myocardial infarction at the age of 40 in her father. Her triglyceride level has varied between 600 and 3000 (normal 20–149 mg/dl) over the years. FPL3 was diagnosed based on genetic testing. She was prescribed fenofibrate and fish oil, and statin was added thereafter. She developed type 2 diabetes and was started on metformin and pioglitazone. She was noted to have hypertension and was treated with amlodipine and lisinopril. She also was found to have Polycystic Ovarian Syndrome (PCOS) based on menstrual irregularities, hirsutism and ultrasound showing multiple ovarian cysts, and was treated with spironolactone. Her most recent labs show triglyceride level of 2400 mg/dl and HbA1c of 8.3. PPARγ gene mutation in FPL3 leads to insulin resistance and hence patients often develop hypertriglyceridemia, type 2 diabetes, PCOS and hypertension. In terms of treatment options, we are still limited to pioglitazone, metformin, statins and fish oil. Often these are not sufficient in addressing the complexity of metabolic derangements in these patients who have an increased risk of cardiovascular events at a young age. Further research about agents targeting this gene in particular would be beneficial. 1. Agarwal et al. A novel heterozygous mutation in peroxisome proliferator-activated receptor-gamma gene in a patient with familial partial lipodystrophy. J Clin Endocrinol Metab. 2002 Jan; 87(1):408–411. 2. Garg A. Lipodystrophies: Genetic and Acquired Body Fat Disorders. J Clin Endocrinol Metab. 2011;96(11): 3313–3325.

PSIX-34 Omega-3 fatty acids are more likely to cause C2C12 cell adipose deposition and decomposition compared with palmitic acid

It is usually considered that n-3 polyunsaturated fatty acids (n-3 PUFAs) to be beneficial to health and are widely provided as a dietary supplement during pregnancy. However, the risk of n-3 PUFAs overdosage on fetal development has rarely been studied. The objective of this study is to explore the effect of different concentrations of n-3 PUFAs [docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA)] and a saturated fatty acid (palmitic acid) on the differentiation of C2C12 myoblasts. Oil Red O staining showed that n-3 PUFAs are more likely to cause adipogenesis. C2C12 cells were induced apoptosis when the concentration of EPA and DHA is 75µM, respectively. The expression of PPARγ gene expression in the EPA+DHA 50μM group was significantly increased, while in the palmitic acid treatment, the gene expression of AP2 (in the ≥10 μM groups) and C/EBPβ (50 μM) were significantly reduced (P &lt; 0.05). Among the lipolysis related genes, HSL increased significantly in the EPA+DHA groups (≥ 25 μM), while MGL decreased significantly in both the EPA+DHA groups (≥ 10 μM) and palmitic acid group (≥ 100 μM) (P &lt; 0.05). In the EPA+DHA groups, the expression of myogenic genes Mrf4 (≥10 μM), MyoD (≥50 μM), and MyoG (≥25 μM) were significantly reduced (P &lt; 0.05). Gene expression of Myf5 decreased significantly when the concentration of palmitic acid was higher than 100 μM (P &lt; 0.05). Among thermogenesis-related genes, in the EPA+DHA treatment, Prdm16 increased (≥50 μM), and UCP3 decreased significantly (≥50 μM)(P &lt; 0.05). After palmitic acid administration, Prdm16 (≥50 μM) and UCP1 (≥100 μM) was significantly reduced, and UCP3 was significantly increased only in the highest concentration group (150 μM) (P &lt; 0.05). In short, compared to palmitic acid, the treatment of EPA+DHA in the differentiation culture of C2C12 cells can enhance the deposition and decomposition of adipose in a lower concentration.

Mechanisms by Which Membrane and Nuclear ER Alpha Inhibit Adipogenesis in Cells Isolated From Female Mice

Mesenchymal stem cells can differentiate into mature chondrocytes, osteoblasts, and adipocytes. Excessive and dysfunctional visceral adipocytes increase upon menopause and importantly contribute to altered metabolism in postmenopausal women. We previously showed both plasma membrane and nuclear estrogen receptors alpha (ERα) with endogenous estrogen are required to suppress adipogenesis in vivo. Here we determined mechanisms by which these liganded ER pools collaborate to inhibit the peroxisome proliferator-activated gamma (PPARγ) gene and subsequent progenitor differentiation. In 3T3-L1 pre-adipocytes and adipose-derived stem cells (ADSC), membrane ERα signaled through phosphatidylinositol 3-kinase (PI3K)-protein kinase B (AKT) to enhance ERα nuclear localization, importantly at the PPARγ gene promoter. AKT also increased overall abundance and recruitment of co-repressors GATA3, β-catenin, and TCF4 to the PPARγ promoter. Membrane ERα signaling additionally enhanced wingless-integrated (Wnt)1 and 10b expression. The components of the repressor complex were required for estrogen to inhibit rosiglitazone-induced differentiation of ADSC and 3T3-L1 cells to mature adipocytes. These mechanisms whereby ER cellular pools collaborate to inhibit gene expression limit progenitor differentiation to mature adipocytes.

Increasing Fat Deposition via Upregulates the Transcription of Peroxisome Proliferator-Activated Receptor Gamma (PPARγ) in Native Crossbred Chicken

Background: Crossbreeding using exotic breeds is usually employed to improve the growth characteristics of indigenous chickens. This mating not only provides growth but affect adversely to fat deposition as well. We studied the growth, abdominal, subcutaneous and intramuscular fat and mRNA expression of peroxisome proliferator-activated receptor (PPAR) α and PPARγ in adipose and muscle tissues of four chicken breeds [Chee breed (CH) (100% Thai native chicken), Kaimook e-san1 (KM1; 50% CH background), Kaimook e-san2 (KM2; 25% CH background), and broiler (BR)]. This study was aim to study role of PPARs on fat deposition in native crossbred chicken.Results: The BR chickens had higher abdominal fat than other breeds (P<0.05) and the KM2 had an abdominal fat percentage higher than KM1 and CH respectively (P<0.05). The intramuscular fat (IMF) of BR was greater than KM1 and CH (P<0.05). In adipose tissue, PPARα transcription expression was different among the chicken breeds. However, there were breed differences in PPARγ gene expression. Study of abdominal fat PPARγ gene expression showed the BR breed, KM1, and KM2 breed significantly greater (P<0.05) than CH. In 8 to 12 weeks of age, the result shows that the PPARγ expression of the CH breed is less than (P<0.05) KM2. The result of PPARs expression in muscle tissue was similar result in adipose tissue.Conclusion: Crossbreeding improved the growth of the Thai native breed, there was also a corresponding increase in carcass fatness. However, there appears to be a relationship between PPARγ expression and fat deposition traits. therefore, PPARγ activity plays a key role in lipid accumulation by up-regulation.

Pharmacogenetic Aspects of Type 2 Diabetes Treatment

In this article, we analyze the role of different variants of the KCNJ11, TCF7L2, SLC22A1, SLC22A3, CYP2C9, CYP2C8, PPARγ genes polymorphisms in efficacy of diabetes mellitus pharmacotherapy. T allele of the KCNJ11 rs2285676 gene polymorphism and G allele of KCNJ11 rs5218 gene polymorphism are associated with the response to IDPP-4 therapy; the presence of KCNJ11 gene rs5210 polymorphism A allele is a predictor of poor response. The effect of rs7903146 polymorphism of TCF7L2 gene was evaluated on the response to treatment of patients taking linagliptin. Linagliptin significantly reduced HbA1c levels for all three rs7903146 genotypes (CC: –0.82 %; CT: –0.77 %; TT: –0.57 %). A significantly smaller effect of therapy was observed with the genotype with ТТ. The rs622342 polymorphism of SLC22A1 gene was studied in effectiveness of metformin. The researches demonstrated that carriers of variant AA had an average decrease of HbA1c of 0.53 %, heterozygous – decrease of 0.32 %, and carriers of a minor variant of SS had an increase of 0.2 % in the level of HbA1c. A significant effect of CYP2C9 polymorphisms on the pharmacokinetic parameters of PSM was noted. When studying the kinetics of glibenclamide, it was found that carriage of the allele *2 significantly reduces glibenclamide metabolism: homozygous carriers had clearance 90 % lower than homozygous carriers of the wild variant. The studies confirmed the association of the allelic variants of Thr394Thr and Gly482Ser of PPARγ gene with higher efficacy of the rosiglitazone. The data obtained from the analysis of the association of the Pro12Ala polymorphism of PPARγ gene and the response to therapy is contradictory. Thus the personalized approach, based on the knowledge of polymorphism options, will allow choosing the most effective drug with transparent kinetics for each individual patient.