scholarly journals Ciglitazone Increases Adiponectin and Adipogenic Gene Expression in Bovine Skeletal Muscle Satellite Cells

2020 ◽  
Author(s):  
Junfang Zhang ◽  
Yan Yan ◽  
Jianfu Sun ◽  
Ying Wang ◽  
Yan Cui ◽  
...  
Keyword(s):  
Gene Expression ◽  
Signaling Pathway ◽  
Satellite Cells ◽  
Adipocyte Differentiation ◽  
Myogenic Differentiation ◽  
Adiponectin Concentration ◽  
Myogenic Regulatory Factor ◽  
Alcoholic Fatty Liver ◽  
Differentiation Medium ◽  
Adipogenic Gene

Abstract Background: Ciglitazone is a member of the thiazolidinedione (TZD) family, and specifically binds to peroxisome proliferator-activated receptor-γ (PPARγ) , thereby promoting adipocyte differentiation. We hypothesized that ciglitazone as a PPARγ ligand in the absence of an adipocyte differentiation cocktail would increase adiponectin and adipogenic gene expression in bovine satellite cells (BSC).Methods: Muscle-derived bovine satellite cells were isolated from six, 18-month-old Yanbian Yellow Cattle. The BSC were cultured for 96 h in differentiation medium containing 5 µM ciglitazone (CL), 10 µM ciglitazone (CM), or 20 µM ciglitazone (CH). Control (CON) BSC were cultured only in differentiation medium (containing 2% horse serum).Results: The presence of myogenin, desmin, and paired box 7 (Pax7) proteins were confirmed in the BSC by immunofluorescence staining. The CL, CM, and CH treatments produced higher concentrations of triacylglycerol and lipid droplet accumulation in myotubes than those of the CON treatment. Ciglitazone treatments significantly increased the relative expression of PPARγ , CCAAT/enhancer-binding protein alpha (C/EBPα), C/EBPβ, fatty acid synthase stearoyl-CoA desaturase (SCD) and perilipin 2. Ciglitazone treatments increased gene expression of paired box 3 (Pax3) and Pax7 and decreased expression of myogenic differentiation-1, myogenin, myogenic regulatory factor-5 (MRF5), and MYF4 (P < 0.01). Adiponectin concentration caused by ciglitazone treatments was significantly greater than CON (P < 0.01). RNA sequencing showed that 281 differentially expressed genes (DEGs) were found in the treatments of ciglitazone. DEGs gene ontology (GO) analysis showed that the top 10 GO enrichment significantly changed the biological processes such as protein trimerization, negative regulation of cell proliferation, adipocytes differentiation, and cellular response to external stimulus. KEGG pathway analysis showed that DEGs were involved into p53 signaling pathway, PPAR signaling pathway, biosynthesis of amino acids, TNF signaling pathway, non-alcoholic fatty liver disease (NAFLD), PI3K-Akt signaling pathway, and Wnt signaling pathway.Conclusion: These results indicate that ciglitazone acts as PPARγ agonist, effectively increasing the adiponectin concentration and adipogenic gene expression, and stimulating the conversion of BSC to adipocyte-like cells in the absence of adipocyte differentiation cocktail.

scholarly journals PSXIV-32 Oleic acid stimulate the formation of adipocyte-like cells from bovine satellite cells via G Protein Coupled Receptor 43 and CCAAT/Enhancer Binding Protein Beta

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 308-309
Author(s):  
Jongkyoo Kim ◽  
Ki Yong Chung ◽  
Bradley J Johnson ◽  
Stephen B Smith
Keyword(s):  
Gene Expression ◽  
Oleic Acid ◽  
Satellite Cells ◽  
Myogenic Differentiation ◽  
Adipogenic Differentiation ◽  
Horse Serum ◽  
Enhancer Binding Protein ◽  
Mrna Gene ◽  
Fetal Bovine ◽  
Mrna Gene Expression

Abstract Numerous physiological and pathological processes are controlled by free fatty acids (FFAs), which act as a signaling molecule in mammals. We hypothesized that oleic acid (Ole) may stimulate the formation of satellite cell-derived intramuscular adipose tissue. The objective of the current study was to determine the effect of Ole on GPR43 and factors related to the adipogenic differentiation of bovine satellite cells. Bovine satellite cells were isolated from the semimembranosus of two 14-month-old crossbreed steers. The isolated muscle satellite cells were incubated in Dulbecco’s Modified Eagle’s Medium (DMEM) solution with 10% Fetal Bovine Serum. Upon reaching 80 to 90% confluence, the growth medium was replaced with differentiation medium composed of DMEM and 2% horse serum, 10μg/mL insulin, 10μg/mL hydrocortisone, 10μM ciglitizone, and 1×antibiotic-antimycotic with dose of: 0, 1, 10, 100, or 500 μM of oleic acid (Ole). Addition of Ole on BSC induced transdifferentiation of myogenic lineage into adipocyte-like cells which formed lipid droplets within cells. Use of 100 μM and 500 μM Ole doses tended to result in a greater (P &lt; 0.1) amount of mRNA gene expression of C/EBPβ compared to all other doses. This might suppress myogenic differentiation. Expression of PPARγ was not altered (P &gt; 0.1) by treatment. The addition of 100 μM and 500 μM upregulated (P &lt; 0.05) mRNA gene expression of GPR43 and 100 μM of Ole increased protein level of GPR43 (P &lt; 0.05) and phosphorylated AMPKα (P &lt; 0.05).

Skeletal Muscle Regeneration: MSC versus Satellite Cells

2008 ◽  
Vol 139 (2_suppl) ◽  
pp. P86-P86
Author(s):  
Jens Stern-Straeter ◽  
Juritz Stephanie ◽  
Gregor Bran ◽  
Frank Riedel ◽  
Haneen Sadick ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Stem Cells ◽  
Satellite Cell ◽  
Cell Cultures ◽  
Muscle Tissue ◽  
Satellite Cells ◽  
Myogenic Differentiation ◽  
Culture Conditions ◽  
Muscle Biopsies

Problem Differentiating stem cells into the myogenic linage in order to create functional muscle tissue is a challenging endeavour. In this work, adipose-derived mesenchymal stem cells (MSC) and satellite cells derived from muscle biopsies were compared regarding proliferation and myogenic differentiation potential under standardized cell culture conditions. This data was obtained in order to discover the most promising type of stem cell for regeneration of muscle tissue and to determine the optimal culture conditions for later clinical use. Methods Human MSC were isolated from adipose tissue, and primary human skeletal myoblasts were extracted from muscle biopsies by enzymatic digestion. Proliferation was analysed using the AlamarBlue® assay. Gene expression of marker genes – such as Myogenin, Myo D, Myf 5 and MHC – were analysed by RT-PCR. Immunostainings against desmin and sarcomeric-actin were performed as differentiation markers. Results MSC cell cultures showed a greater proliferation rate compared with satellite cell cultures. In both stem cell cultures, myogenic differentiation/heritage could be verified by immunostainings against the muscle-specific marker desmin. Gene expression and protein analysis revealed a more stable differentiation of human satellite cell cultures. Conclusion Characterization of both human MSC cultures and satellite cell cultures – and thereby an understanding of myogenesis – might lead to their clinical usage in skeletal muscle tissue engineering. The results in this study appear to indicate that human satellite cell cultures have a more stable differentiation under in vitro conditions and that they might offer a greater potential for skeletal muscle tissue engineering purposes. Significance Our study contributes to the understanding of myogenic differentiation of MSC and satellite cells and helps to improve culture systems for later clinical utilization.

scholarly journals CDR1as related miRNA-mRNA networks in differentiating goat skeletal muscle satellite cells

2020 ◽  
Author(s):  
Bismark Kyei ◽  
Li Li ◽  
Liu Yang ◽  
Siyuan Zhan ◽  
Juntao Li ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Signaling Pathway ◽  
Satellite Cells ◽  
Muscle Development ◽  
Myogenic Differentiation ◽  
Mapk Signaling ◽  
Circular Rnas ◽  
Messenger Rnas ◽  
Muscle Satellite Cells ◽  
Skeletal Muscle Satellite Cells

Abstract Background: Myogenesis is a complex process controlled by several coding and non-coding RNAs (ncRNAs) such as circular RNAs (circRNAs) that well-known function as endogenous microRNAs (miRNAs) sponges. Over the past few years, numerous circRNAs have been known and their roles in biological processes have begun to be understood. Cerebellar Degeneration-Related protein 1 antisense (CDR1as), the most spotlighted circRNA as miR-7 sponge that has been blooming circRNAs’ research for a decade, and can potentially sponge several miRNAs in disease and muscle physiology. Nevertheless, the linear-RNAs-differed character that the acute interventions for circRNAs do not affect miRNAs levels, and has retarded the transcriptome-wide discovery of miRNAs sponged by. Therefore, the purpose of this study was to provide the transcriptomic effect of CDR1as during muscle differentiation.Methods: siCDR1as and siDICER1 were transfected into goat skeletal muscle satellite cells (SMSCs). RNA-seq technology and bioinformatics tools were used to analyze genes that are deregulated by siCDR1as and siDICER1. quantitative PCR was used to verify the expression levels of the differentially expressed mRNAs and miRNAs. Results: Here, to systematically identify miRNAs targeting CDR1as, we employed the critical enzyme DICER1 that governs the biogenesis of miRNAs. The deficiency of either DICER1 or CDR1as inhibited myogenic differentiation of SMSCs, and knockdown of DICER1 decreased the expression of CDR1as. Moreover, we screened for the targeted messenger RNAs (mRNAs) and miRNAs in SMSCs transfected with siDICER1 or siCDR1as respectively and found out that some well-known muscle-related pathways such as phosphoinositide 3-kinase (PI3K)-AKT signaling pathway, Rap1 signaling pathway, and MAPK signaling pathway were enriched in all groups. Further, regarding the miRNAs identified in siDICER1 and siCDR1as together with the sequence complementary information, we identified 11 miRNAs including miR-1, miR-206, and miR-27a-5p which are more likely to be novel targets for CDR1as. Conclusion: In summary, our study provides a perspective on the potential functions and relationship between CDR1as and DICER1 during muscle development.

scholarly journals Oleic acid in the absence of a PPARγ agonist increases adipogenic gene expression in bovine muscle satellite cells1

2019 ◽  
Vol 97 (10) ◽  
pp. 4114-4123 ◽  
Author(s):  
Xiang Z Li ◽  
Yan Yan ◽  
Jun F Zhang ◽  
Jian F Sun ◽  
Bin Sun ◽  
...  
Keyword(s):  
Gene Expression ◽  
Oleic Acid ◽  
Myogenic Differentiation ◽  
Binding Protein ◽  
Regulatory Element ◽  
Horse Serum ◽  
Peroxisome Proliferator ◽  
Bovine Muscle ◽  
Pparγ Agonist ◽  
Adipogenic Gene

Abstract We hypothesized that oleic acid (OA) in the absence of a thiazolidinedione (i.e., a synthetic peroxisome proliferator-activated receptorγ [PPARγ] agonist) would increase adipogenic gene expression in bovine muscle satellite cells (BSC). The BSC were cultured in differentiation medium containing 10 µM ciglitazone (CI), 100 µM OA, or 100 µM OA plus 10 µM CI (CI-OA). Control (CON) BSC were cultured only in differentiation media (containing 2% horse serum). The presence of myogenin, desmin, and paired box 7 proteins was confirmed in the BSC by immunofluorescence staining, demonstrating that we had isolated myogenic cells. The OA BSC had lesser paired box 3 (Pax3) and myogenic differentiation 1 expression but greater Pax7 and mygogenin (MYOG) expression (P < 0.05), than the CON BSC. The CI BSC had greater Pax3, Pax7, and MYOG expression than CON BSC (P < 0.05), suggesting that CI would promote BSC myogenesis under pro-myogenic conditions (i.e., when cultured with horse serum). However, both the OA and CI treatments upregulated the expression of PPARγ, CCAAT/enhancer-binding protein alpha (C/EBPα) and C/EBPß, sterol regulatory element-binding protein 1, lipoprotein lipase, and glycerol-3-phosphate acyltransferase 3 gene expression, as well as media adiponectin concentration (P < 0.05). The CI, OA, and CI-OA treatments also increased triacylglycerol and lipid droplet accumulation, in spite of upregulation (relative to CON BSC) of adenosine monophosphate-activated protein kinase alpha-1, perilipin 2 (PLIN2), and PLIN3 in BSC and downregulation of G protein-coupled protein receptor 43, acyl-CoA synthetase long chain family member 3, and stearoyl-CoA desaturase (P < 0.05). These results indicate that OA in the absence of a synthetic PPARγ agonist can effectively increase adipogenic gene expression in BSC.

scholarly journals Reduced Adipogenic Gene Expression in Thigh Adipose Tissue Precedes Human Immunodeficiency Virus-Associated Lipoatrophy

2008 ◽  
Vol 93 (3) ◽  
pp. 959-966 ◽  
Author(s):  
Mario Kratz ◽  
Jonathan Q. Purnell ◽  
Patricia A. Breen ◽  
Katherine K. Thomas ◽  
Kristina M. Utzschneider ◽  
...  
Keyword(s):  
Gene Expression ◽  
Adipose Tissue ◽  
Insulin Sensitivity ◽  
Adipocyte Differentiation ◽  
Plasma Lipids ◽  
Antiretroviral Drugs ◽  
Metabolic Complications ◽  
Prospective Longitudinal Study ◽  
Adipogenic Gene ◽  
Prospective Longitudinal

Abstract Context: The expression of adipogenic genes in sc adipose tissue has been reported to be lower among patients with HIV-associated lipoatrophy than HIV-uninfected controls. It is unclear whether this is a result or cause of lipoatrophy. Objective: The objective of the study was to investigate the temporal relationships among changes in adipogenic gene expression in sc adipose tissue and changes in body fat distribution and metabolic complications in HIV-infected subjects on antiretroviral therapy. Design: This was a prospective longitudinal study. Setting: The study was conducted at HIV clinics in Seattle, Washington. Participants: The study population included 31 HIV-infected and 12 control subjects. Interventions: Subjects were followed up for 12 months after they initiated or modified their existing antiretroviral regimen. Main Outcome Measures: Changes in body composition, plasma lipids, insulin sensitivity, and gene expression in sc abdominal and thigh adipose tissue. Results: Subjects who developed lipoatrophy (n = 10) had elevated fasting triglycerides [3.16 (sd 2.79) mmol/liter] and reduced insulin sensitivity as measured by frequently sampled iv glucose tolerance test [1.89 (sd 1.27) × 10−4 min−1/μU·ml] after 12 months, whereas those without lipoatrophy (n = 21) did not show any metabolic complications [triglycerides 1.32 (sd 0.58) mmol/liter, P = 0.01 vs. lipoatrophy; insulin sensitivity 3.52 (sd 1.91) × 10−4 min−1/μU·ml, P = 0.01 vs. lipoatrophy]. In subjects developing lipoatrophy, the expression of genes involved in adipocyte differentiation, lipid uptake, and local cortisol production in thigh adipose tissue was significantly reduced already at the 2-month visit, several months before any loss of extremity fat mass was evident. Conclusions: In HIV-infected subjects, lipoatrophy is associated with elevated fasting triglycerides and insulin resistance and might be caused by a direct or indirect effect of antiretroviral drugs on sc adipocyte differentiation.

scholarly journals Effect of Temperature and Selection for Growth on Intracellular Lipid Accumulation and Adipogenic Gene Expression in Turkey Pectoralis Major Muscle Satellite Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Jiahui Xu ◽  
Gale M. Strasburg ◽  
Kent M. Reed ◽  
Sandra G. Velleman
Keyword(s):  
Gene Expression ◽  
Thermal Stress ◽  
Satellite Cells ◽  
Lipid Accumulation ◽  
Lipid Synthesis ◽  
Pectoralis Major ◽  
Effect Of Temperature ◽  
Control Line ◽  
Intracellular Lipid ◽  
Adipogenic Gene

As multipotential stem cells, satellite cells (SCs) have the potential to express adipogenic genes resulting in lipid synthesis with thermal stress. The present study determined the effect of temperature on intracellular lipid synthesis and adipogenic gene expression in SCs isolated from the pectoralis major (p. major) muscle of 7-day-old fast-growing modern commercial (NC) turkeys compared to SCs from unselected slower-growing turkeys [Randombred Control Line 2 (RBC2)]. Since proliferating and differentiating SCs have different responses to thermal stress, three incubation strategies were used: (1) SCs proliferated at the control temperature of 38°C and differentiated at 43° or 33°C; (2) SCs proliferated at 43° or 33°C and differentiated at 38°C; or (3) SCs both proliferated and differentiated at 43°, 38°, or 33°C. During proliferation, lipid accumulation increased at 43°C and decreased at 33°C with the NC line showing greater variation than the RBC2 line. During proliferation at 43°C, peroxisome proliferator-activated receptor-γ (PPARγ) and neuropeptide-Y (NPY) expression was reduced to a greater extent in the NC line than the RBC2 line. At 33°C, expression of PPARγ, NPY, and CCAAT/enhancer-binding protein-β (C/EBPβ) was upregulated, but only in the RBC2 line. During differentiation, both lines showed greater changes in lipid accumulation and in C/EBPβ and NPY expression if the thermal challenge was initiated during proliferation. These data suggest that adipogenic gene expression is more responsive to thermal challenge in proliferating SCs than in differentiating SCs, and that growth-selection has increased temperature sensitivity of SCs, which may significantly affect breast muscle structure and composition.

scholarly journals A214 BARIATRIC SURGERY PATIENTS WITH NON-ALCOHOLIC FATTY LIVER DISEASE HAVE DIFFERENT VISCERAL ADIPOSE TISSUE GENE EXPRESSION COMPARED TO THOSE WITH NORMAL LIVER

2021 ◽  
Vol 4 (Supplement_1) ◽  
pp. 245-247
Author(s):  
S Keshavjee ◽  
J Yadav ◽  
K Schwenger ◽  
S Fischer ◽  
T D Jackson ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bariatric Surgery ◽  
Adipose Tissue ◽  
Visceral Adipose Tissue ◽  
Inflammatory Markers ◽  
Fatty Liver Disease ◽  
Alcoholic Fatty Liver ◽  
Visceral Adipose ◽  
Healthy Liver ◽  
Alcoholic Fatty Liver Disease

Abstract Background Non-alcoholic fatty liver disease (NAFLD) includes simple steatosis (SS) and nonalcoholic steatohepatitis (NASH). It affects 74–98% of individuals with morbid obesity undergoing bariatric surgery (BSX). Among several factors contributing to NAFLD pathogenesis, adipokines secreted by visceral adipose tissue (VAT) can play a role by regulating glucose/lipid metabolism and inflammation. Aims This study aims to determine if visceral adipose tissue adipokine and cytokine gene expression are associated with NAFLD (SS and NASH) at the time of BSX. Methods Patients were recruited from the Toronto Western Hospital Bariatric Clinic. Demographic data was recorded. The VAT and liver biopsies were collected at the time of bariatric surgery. VAT adipokines and other mediators were assessed by RT-PCR and included markers of thermogenic capacity, inflammation, fibrosis, adipokines, and others. Liver histology was assessed by a pathologist using the Brunt system and individuals were diagnosed as either SS, NASH, or having a healthy liver (HL). Blood samples were collected pre-BSX to measure liver and metabolic syndrome related parameters, including HOMA-IR, HbA1c, liver enzymes, and lipid profile. Anthropometry was also assessed. Groups were compared using Kruskal-Wallis test followed by Wilcoxon ranked sum, or chi-square and Fisher’s exact test as necessary. Data was considered to be statistically significant with a p-value less than 0.05. Results We are presenting data on 126 patients, 80.2% females with a median age of 49 and a body mass index (BMI) of 46.9. Fifty-seven patients had SS, 34 had NASH and 35 had a healthy liver (HL). BMI, age, and sex did not differ between the three groups. First, we found that those with NASH had significantly higher VAT expression of fibrosis (Loxl2), inflammation (CCL4 and TGFb1) and proliferation markers (E2F1) and significantly lower expression of adipokines (TNFa and resistin) compared to HL. Also, we found that SS had significantly higher fibrosis (Col3a1, Col6a1, Loxl2, CD9 and Acta2), inflammation (Nox2, TGFb1, IFNg and Clec10a), browning (PPARa, PPARg and Glut1) and proliferation (E2F1) marker expression compared to HL. Conclusions Results show that there is a significant difference in the expression pattern of VAT fibrotic and inflammatory markers between HL, SS and NASH patients. The observed increase of inflammatory markers in NAFLD is in line with prior research outlining the ability of inflammatory mediators from VAT to contribute to liver pathology via portal circulation. The relationship between VAT characteristics and NAFLD are important in understanding the widespread metabolic effects of obesity. Funding Agencies CIHRCanadian Liver foundation

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