scholarly journals De-novo and depot-specific androgen production in human adipose tissue - a source of hyperandrogenism in women with obesity

Obesity Facts ◽  
2022 ◽  
Author(s):  
Isabel Viola Wagner ◽  
Iuliia Savchuk ◽  
Lena Sahlin ◽  
Alexandra Kulle ◽  
Nora Klöting ◽  
...  
Keyword(s):  
Cell Culture ◽  
Adipose Tissue ◽  
De Novo ◽  
Elevated Serum ◽  
Cell Culture Supernatant ◽  
Obese Women ◽  
Steroidogenic Enzymes ◽  
Androgen Synthesis ◽  
Gene And Protein Expression ◽  
Androgen Levels

Introduction: Obesity in women is often associated with hyperandrogenism but the role of adipose tissue (AT) in androgen synthesis remains unclear. Therefore, we studied whether AT could be a source of androgens promoting hyperandrogenism. Methods: Subcutaneous and visceral AT was collected from lean and obese women. Androgen levels were evaluated in serum, AT and cell culture supernatant. Gene and protein expression of steroidogenic enzymes were determined. Results: Obese subjects had elevated serum androgen levels, which reduced after weight loss. Androgens were measurable in AT and in cell culture supernatants of adipocytes. Steroids were higher in AT from obese women, with the highest difference for testosterone in visceral AT (+7.9 fold, p=0.032). Steroidogenic enzymes were expressed in human AT with depot-specific differences. Obese women showed a significantly higher expression of genes of the backdoor pathway and of CYP19 in visceral AT. Conclusion: The whole steroidogenic machinery of the classical and backdoor pathways of steroidogenesis, and the capacity for androgen biosynthesis, were found in both AT depots and cultured adipocytes. Therefore, we hypothesize that AT is a de novo site of androgen production and the backdoor pathway of steroidogenesis might be a new pathomechanism for hyperandrogenism in women with obesity.

scholarly journals Downregulation of de Novo Fatty Acid Synthesis in Subcutaneous Adipose Tissue of Moderately Obese Women

2015 ◽  
Vol 16 (12) ◽  
pp. 29911-29922 ◽  
Author(s):  
Esther Guiu-Jurado ◽  
Teresa Auguet ◽  
Alba Berlanga ◽  
Gemma Aragonès ◽  
Carmen Aguilar ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Fatty Acid ◽  
Fatty Acid Synthesis ◽  
Subcutaneous Adipose Tissue ◽  
De Novo ◽  
Acid Synthesis ◽  
Obese Women ◽  
Subcutaneous Adipose

Select de novo Gene and Protein Expression During Renal Epithelial Cell Culture in Rotating Wall Vessels is Shear Stress Dependent

1999 ◽  
Vol 168 (1) ◽  
pp. 77-89 ◽  
Author(s):  
J.H. Kaysen *, § , W.C. Campbell ◽  
R.R. Majewski ◽  
F.O. Goda ◽  
G.L. Navar ◽  
F.C. Lewis *, † , T.J. Goodwin
Keyword(s):  
Cell Culture ◽  
Shear Stress ◽  
Epithelial Cell ◽  
De Novo ◽  
Epithelial Cell Culture ◽  
Renal Epithelial Cell ◽  
Rotating Wall ◽  
De Novo Gene ◽  
Gene And Protein Expression ◽  
Stress Dependent

scholarly journals Resistin Is More Abundant in Liver Than Adipose Tissue and Is Not Up-Regulated by Lipopolysaccharide

2009 ◽  
Vol 94 (8) ◽  
pp. 3051-3057 ◽  
Author(s):  
Ewa Szalowska ◽  
Marieke G. L. Elferink ◽  
Annemiek Hoek ◽  
Geny M. M. Groothuis ◽  
Roel J. Vonk
Keyword(s):  
Adipose Tissue ◽  
Protein Expression ◽  
Liver Tissue ◽  
Inflammatory Markers ◽  
Human Liver ◽  
Elevated Serum ◽  
Human Liver Tissue ◽  
Gene And Protein Expression ◽  
Total Body ◽  
High Level

Context: Resistin is an adipokine correlated with inflammatory markers and is predictive for cardiovascular diseases. There is evidence that serum resistin levels are elevated in obese patients; however, the role of resistin in insulin resistance and type 2 diabetes remains controversial. Objective: We addressed the question of whether inflammation may induce expression of resistin in organs involved in regulation of total body energy metabolism, such as liver and adipose tissue (AT). Methods: Human liver tissue, sc AT, and omentum were cultured in the absence/presence of lipopolysaccharide (LPS). The resistin and cytokine mRNA and protein expression levels were determined by real-time PCR, ELISA, and Multiplex Technology, respectively. The localization of resistin in human liver was analyzed by immunohistochemistry. Results: Resistin gene and protein expression was significantly higher in liver than in AT. Exposure of human AT and liver tissue in culture to LPS did not alter resistin concentration; however, concentrations of IL-1β, IL-6, and TNFα were significantly increased in these tissues. In liver, resistin colocalizes with markers for Kupffer cells, for a subset of endothelial and fibroblast-like cells. Conclusions: High level of resistin gene and protein expression in liver compared to AT implies that resistin should not be considered only as an adipokine in humans. LPS-induced inflammation does not affect resistin protein synthesis in human liver and AT. This suggests that elevated serum resistin levels are not indicative for inflammation of AT or liver in a manner similar to known inflammatory markers such as IL-1β, IL-6, or TNFα.

Separation of Extracellular Vesicles by DNA-Directed Immunocapturing Followed by Enzymatic Release

2020 ◽  
Author(s):  
Dario Brambilla ◽  
Laura Sola ◽  
Elisa Chiodi ◽  
Natasa Zarovni ◽  
Diogo Fortunato ◽  
...  
Keyword(s):  
Cell Culture ◽  
Extracellular Vesicles ◽  
Culture Media ◽  
Biological Fluids ◽  
Imaging Techniques ◽  
Cell Communication ◽  
Disease Diagnosis ◽  
Cell Culture Supernatant ◽  
Transmission Electron ◽  
Downstream Analysis

Extracellular vesicles (EVs) have attracted great interest among researchers due to their role in cell-cell communication, disease diagnosis, and drug delivery. In spite of their potential in the medical field, there is no consensus on the best method for separating microvesicles from cell culture supernatant and complex biological fluids. Obtaining a good recovery yield and preserving physical characteristics is critical for the diagnostic and therapeutic use of EVs. The separation is made complex by the fact that blood and cell culture media, contain a large number of nanoparticles in the same size range. Methods that exploit immunoaffinity capture provide high purity samples and overcome the issues of currently used separation methods. However, the release of captured nanovesicles requires harsh conditions that hinder their use in certain types of downstream analysis. Herein, a novel capture and release approach for small extracellular vesicles (sEVs), based on DNAdirected immobilization of antiCD63 antibody is presented. The flexible DNAlinker increases the capture efficiency and allows releasing of EVs by exploiting the endonucleasic activity of DNAse I. This separation protocol works under mild conditions, enabling the release of intact vesicles that can be successfully analyzed by imaging techniques. In this article sEVs recovered from plasma were characterized by established techniques for EVs analysis including nanoparticle tracking and transmission electron microscopy.<br>

Continuous purification of antibodies from cell culture supernatant with aqueous two-phase systems: From concept to process

2013 ◽  
Vol 8 (3) ◽  
pp. 352-362 ◽  
Author(s):  
Paula A. J. Rosa ◽  
Ana M. Azevedo ◽  
S. Sommerfeld ◽  
Martina Mutter ◽  
Werner Bäcker ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Supernatant ◽  
Cell Culture Supernatant ◽  
Two Phase ◽  
Aqueous Two Phase Systems ◽  
Phase Systems

scholarly journals Expression Signatures of microRNAs and Their Targeted Pathways in the Adipose Tissue of Chickens during the Transition from Embryonic to Post-Hatch Development

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 196
Author(s):  
Julie A. Hicks ◽  
Hsiao-Ching Liu
Keyword(s):  
Energy Source ◽  
Adipose Tissue ◽  
De Novo ◽  
Lipid Storage ◽  
Storage Site ◽  
Metabolic Processes ◽  
Metabolic Switch ◽  
Regulatory Pathways ◽  
Main Site ◽  
Real Time Quantitative Pcr

As the chick transitions from embryonic to post-hatching life, its metabolism must quickly undergo a dramatic switch in its major energy source. The chick embryo derives most of its energy from the yolk, a lipid-rich/carbohydrate-poor source. Upon hatching, the chick’s metabolism must then be able to utilize a lipid-poor/carbohydrate-rich source (feed) as its main form of energy. We recently found that a number of hepatically-expressed microRNAs (miRNAs) help facilitate this shift in metabolic processes in the chick liver, the main site of lipogenesis. While adipose tissue was initially thought to mainly serve as a lipid storage site, it is now known to carry many metabolic, endocrine, and immunological functions. Therefore, it would be expected that adipose tissue is also an important factor in the metabolic switch. To that end, we used next generation sequencing (NGS) and real-time quantitative PCR (RT-qPCR) to generate miRNome and transcriptome signatures of the adipose tissue during the transition from late embryonic to early post-hatch development. As adipose tissue is well known to produce inflammatory and other immune factors, we used SPF white leghorns to generate the initial miRNome and transcriptome signatures to minimize complications from external factors (e.g., pathogenic infections) and ensure the identification of bona fide switch-associated miRNAs and transcripts. We then examined their expression signatures in the adipose tissue of broilers (Ross 708). Using E18 embryos as representative of pre-switching metabolism and D3 chicks as a representative of post-switching metabolism, we identified a group of miRNAs which work concordantly to regulate a diverse but interconnected group of developmental, immune and metabolic processes in the adipose tissue during the metabolic switch. Network mapping suggests that during the first days post-hatch, despite the consumption of feed, the chick is still heavily reliant upon adipose tissue lipid stores for energy production, and is not yet efficiently using their new energy source for de novo lipid storage. A number of core master regulatory pathways including, circadian rhythm transcriptional regulation and growth hormone (GH) signaling, likely work in concert with miRNAs to maintain an essential balance between adipogenic, lipolytic, developmental, and immunological processes in the adipose tissue during the metabolic switch.

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