A Novel Age-Related Circular RNA Circ-ATXN2 Inhibits Proliferation, Promotes Cell Death and Adipogenesis in Rat Adipose Tissue-Derived Stromal Cells

Adipose tissue-derived stromal cells are promising candidates investigating the stem cell-related treatment. However, their proportion and utility in the human body decline with time, rendering stem cells incompetent to complete repair processes in vivo. The involvement of circRNAs in the aging process is poorly understood. Rat subcutaneous adipose tissue from 10-week-old and 27-month-old rats were used for hematoxylin and eosin (H and E) staining, TUNEL staining, and circRNA sequencing. Rat adipose tissue-derived stromal cells were cultured and overexpressed with circ-ATXN2. Proliferation was examined using xCELLigence real-time cell analysis, EdU staining, and cell cycle assay. Apoptosis was induced by CoCl2 and examined using flow cytometry. RT-PCR assay and Oil Red O staining were used to measure adipogenesis at 48 h and 14 days, respectively. H and E staining showed that the diameter of adipocytes increased; however, the number of cells decreased in old rats. TUNEL staining showed that the proportion of apoptotic cells was increased in old rats. A total of 4,860 and 4,952 circRNAs was detected in young and old rats, respectively. Among them, 67 circRNAs exhibited divergent expression between the two groups (fold change ≥2, p ≤ 0.05), of which 33 were upregulated (49.3%) and 34 were downregulated (50.7%). The proliferation of circ-ATXN2-overexpressing cells decreased significantly in vitro, which was further validated by xCELLigence real-time cell analysis, EdU staining, and cell cycle assay. Overexpression of circ-ATXN2 significantly increased the total apoptotic rate from 5.78 ± 0.46% to 11.97 ± 1.61%, early apoptotic rate from 1.76 ± 0.22% to 5.50 ± 0.66%, and late apoptosis rate from 4.02 ± 0.25% to 6.47 ± 1.06% in adipose tissue-derived stromal cells. Furthermore, in circ-ATXN2-overexpressing cells, RT-PCR assay revealed that the expression levels of adipose differentiation-related genes PPARγ and CEBP/α were increased and the Oil Red O staining assay showed more lipid droplets. Our study revealed the expression profile of circRNAs in the adipose tissue of old rats. We found a novel age-related circular RNA—circ-ATXN2—that inhibits proliferation and promotes cell death and adipogenesis in rat adipose tissue-derived stromal cells.

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Pathogenic Microenvironment from Diabetic–Obese Visceral and Subcutaneous Adipocytes Activating Differentiation of Human Healthy Preadipocytes Increases Intracellular Fat, Effect of the Apocarotenoid Crocetin

Nutrients ◽

10.3390/nu13031032 ◽

2021 ◽

Vol 13 (3) ◽

pp. 1032

Author(s):

Lesgui Alviz ◽

David Tebar-García ◽

Raquel Lopez-Rosa ◽

Eva M. Galan-Moya ◽

Natalia Moratalla-López ◽

...

Keyword(s):

Adipose Tissue ◽

Subcutaneous Adipose Tissue ◽

Bioactive Components ◽

Hypertrophic Growth ◽

Visceral Adipose ◽

Subcutaneous Adipose ◽

Oil Red O Staining ◽

Oil Red O ◽

Excessive Accumulation

In diabetes mellitus type 2 (DM2), developed obesity is referred to as diabesity. Implementation of a healthy diet, such as the Mediterranean, prevents diabesity. Saffron is frequently used in this diet because of its bioactive components, such as crocetin (CCT), exhibit healthful properties. It is well known that obesity, defined as an excessive accumulation of fat, leads to cardiometabolic pathology through adiposopathy or hypertrophic growth of adipose tissue (AT).This is related to an impaired adipogenic process or death of adipocytes by obesogenic signals. We aimed to evaluate the effect of the pathogenic microenvironment and CCT, activating differentiation of healthy preadipocytes (PA). For this, we used human cryopreserved PA from visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) depots obtained from healthy and obese-DM2 donors. We studied the effect of a metabolically detrimental (diabesogenic) environment, generated by obese-DM2 adipocytes from VAT (VdDM) or SAT (SdDM), on the viability and accumulation of intracellular fat of adipocytes differentiated from healthy PA, in the presence or absence of CCT (1 or 10 μM). Intracellular fat was quantified by Oil Red O staining. Cytotoxicity was measured using the MTT assay. Our results showed that diabesogenic conditions induce cytotoxicity and provide a proadipogenic environment only for visceral PA. CCT at 10 μM acted as an antiadipogenic and cytoprotective compound.

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The Cooperation of Adipocytes and Stromal Cells in the Secretion of Prostaglandins by Rat Adipose Tissue Is Not Influenced by Diet

Journal of Nutrition ◽

10.1093/jn/123.7.1203 ◽

1993 ◽

Vol 123 (7) ◽

pp. 1203-1207 ◽

Cited By ~ 2

Author(s):

Minghua Zhang ◽

Dorothy B. Hausman ◽

Gary J. Hausman

Keyword(s):

Adipose Tissue ◽

Stromal Cells ◽

Rat Adipose Tissue

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Dietary sodium restriction exacerbates age-related changes in rat adipose tissue and liver lipogenesis

Metabolism ◽

10.1016/s0026-0495(03)00162-8 ◽

2003 ◽

Vol 52 (8) ◽

pp. 1072-1077 ◽

Cited By ~ 8

Author(s):

A.R Xavier ◽

M.A.R Garófalo ◽

R.H Migliorini ◽

I.C Kettelhut

Keyword(s):

Adipose Tissue ◽

Dietary Sodium ◽

Sodium Restriction ◽

Age Related ◽

Dietary Sodium Restriction ◽

Rat Adipose Tissue ◽

Age Related Changes

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Age related changes in the antilipolytic effects of nicotinic acid in rat adipose tissue

British Journal of Pharmacology ◽

10.1111/j.1476-5381.1983.tb10056.x ◽

1983 ◽

Vol 80 (4) ◽

pp. 663-670 ◽

Cited By ~ 3

Author(s):

L. Caparrotta ◽

G. Fassina ◽

R.M. Gaion ◽

F. Tessari

Keyword(s):

Adipose Tissue ◽

Nicotinic Acid ◽

Age Related ◽

Rat Adipose Tissue ◽

Age Related Changes

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Alterations in rat adipose tissue morphology induced by a low-temperature environment

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1982.242.2.e93 ◽

1982 ◽

Vol 242 (2) ◽

pp. E93-E96 ◽

Cited By ~ 2

Author(s):

W. H. Miller ◽

I. M. Faust

Keyword(s):

Adipose Tissue ◽

Sprague Dawley ◽

High Fat ◽

Fat Depots ◽

High Carbohydrate ◽

Tissue Morphology ◽

Age Related ◽

Sprague Dawley Rats ◽

Temperature Environment ◽

Rat Adipose Tissue

Rats raised in the cold showed an unusual pattern of adipose tissue morphology. Male Sprague-Dawley rats were maintained in a 5 degrees C environment for up to 24 wk and the cellularity of their major adipose depots was determined. Normal age-related increases in adipocyte number were absent in two major fat depots (retroperitoneal and inguinal), whereas there was a supranormal increase in a third (epididymal). This pattern of hyperplasia contrasts sharply with that seen in rats fed highly palatable high-fat or high-carbohydrate diets in which retroperitoneal depots show the most hyperplasia and epididymal pads the least. Such variations of response across depots suggest that the features of adipose tissue responsible for adipocyte proliferation in the various depots may not be homogeneous both in their nature and in their distribution.

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The Initial Cardiac Tissue Response to Cryopreserved Allogeneic Adipose Tissue-Derived Mesenchymal Stromal Cells in Rats with Chronic Ischemic Cardiomyopathy

International Journal of Molecular Sciences ◽

10.3390/ijms222111758 ◽

2021 ◽

Vol 22 (21) ◽

pp. 11758

Author(s):

Bjarke Follin ◽

Cecilie Hoeeg ◽

Lisbeth D. Højgaard ◽

Morten Juhl ◽

Kaya B. Lund ◽

...

Keyword(s):

Adipose Tissue ◽

Gene Transcription ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Ischemic Cardiomyopathy ◽

Cardiac Tissue ◽

Gene Expression Pattern ◽

Saline Group ◽

Tissue Response ◽

Rat Adipose Tissue

Mesenchymal stromal cells have proven capable of improving cardiac pump function in patients with chronic heart failure, yet little is known about their mode of action. The aim of the study was to investigate the short-term effect of cryopreserved allogeneic rat adipose tissue-derived stromal cells (ASC) on cardiac composition, cellular subpopulations, and gene transcription in a rat model of chronic ischemic cardiomyopathy (ICM). Myocardial infarction (MI) was induced by permanent ligation of the left anterior descending coronary artery. After 6 weeks, the rats were treated with ASCs, saline, or no injection, using echo-guided trans-thoracic intramyocardial injections. The cardiac tissue was subsequently collected for analysis of cellular subpopulations and gene transcription 3 and 7 days after treatment. At day 3, an upregulation of genes associated with angiogenesis were present in the ASC group. On day 7, increases in CCR2+ and CD38+ macrophages (p = 0.047 and p = 0.021), as well as in the CD4/CD8 lymphocyte ratio (p = 0.021), were found in the ASC group compared to the saline group. This was supported by an upregulation of genes associated with monocytes/macrophages. In conclusion, ASC treatment initiated an immune response involving monocytes/macrophages and T-cells and induced a gene expression pattern associated with angiogenesis and monocyte/macrophage differentiation.

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Torreya nucifera seed oil improves 3T3-L1 adipocyte differentiation

BMC Complementary Medicine and Therapies ◽

10.1186/s12906-021-03429-5 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Eunbi Koh ◽

Boram Kim ◽

Kyungoh Choi

Keyword(s):

Adipose Tissue ◽

Fatty Acid ◽

Seed Oil ◽

Adipocyte Differentiation ◽

Endocrine Function ◽

Mrna Levels ◽

Fatty Acid Binding ◽

Oil Red O Staining ◽

Intracellular Triglyceride ◽

Oil Red O

Abstract Background Adipose tissue is a critical regulator of lipid storage and endocrine function. Impairment of the recruitment of new adipocytes in the adipose tissue is associated with ectopic fat accumulation, diabetes and insulin resistance. Torreya nucifera, an evergreen conifer that grows in warm temperate climates, has been found to exert beneficial effects against inflammation, infection and diabetes. However, the molecular mechanisms responsible for these effects at the cellular level remain unknown. This study aimed to investigate effects of Torreya nucifera seed oil (TNSO) on 3T3-L1 adipocyte differentiation and its underlying regulatory mechanism. Methods To investigate the effects of TNSO on adipocyte differentiation, 3T3-L1 cells were induced to differentiate for 5 days in the presence of 0.75 μL/mL TNSO. Oil Red O staining and an assay for intracellular triglyceride were performed to determine the extent of lipid accumulation in 3T3-L1 cells. To elucidate the underlying mechanism of TNSO, adipogenic gene expression was analyzed using quantitative real-time PCR. Moreover, we monitored TNSO-derived activation of PPARγ and STAT3 with 3T3-L1 reporter cell lines engineered to secrete Gaussia luciferase upon the interaction of a transcription factor to its DNA binding element. Results Oil Red O staining revealed that TNSO improved the differentiation of 3T3-L1 preadipocytes into mature adipocytes. The mRNA levels of adipogenic genes, including adiponectin, fatty acid synthase (FAS) and adipocyte fatty acid-binding protein (FABP4), were upregulated and intracellular triglyceride levels increased upon TNSO treatment. We also established that adipocyte differentiation was improved by TNSO-derived activation of PPARγ and STAT3. Conclusions Our results suggest that TNSO improves adipocyte differentiation by regulating the activation of adipogenic transcription factors, indicating that it may serve as a potential treatment strategy for adipocyte dysfunction.

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Abstract P120: Increased [ca 2+ ] e Enhances Adipocyte Development In Bone Marrow Stroma

Circulation Research ◽

10.1161/res.109.suppl_1.ap120 ◽

2011 ◽

Vol 109 (suppl_1) ◽

Author(s):

Ryota Hashimoto ◽

Youichi Katoh ◽

Seigo Itoh ◽

Takafumi Iesaki ◽

Hiroyuki Daida ◽

...

Keyword(s):

Bone Marrow ◽

Stromal Cells ◽

Bone Marrow Stromal Cells ◽

Marrow Stromal Cells ◽

Bone Marrow Stroma ◽

Age Related ◽

Marrow Stroma ◽

High Concentrations ◽

Adipocyte Development ◽

Oil Red O

Background: Bone marrow stroma contains adipocytes, osteoblasts, and lymphohematopoietic donor cells. With age, fatty marrow gradually predominates in bone marrow stroma and is a factor underlying age-related fracture and anemia. Thus, it is important to understand the mechanism of adipocyte development in bone marrow stroma. Bone marrow Ca 2+ levels can reach high concentrations of 8 to 40 mM, while circulating plasma Ca 2+ levels normally range from 2.3 to 2.6 mM. However, the effects of a high extracellular calcium concentration ([Ca 2+ ] e ) on adipocyte development in bone marrow stroma remain largely unknown. Methods and Results: We studied the effects of high [Ca 2+ ] e on adipocyte development in bone marrow stroma. First, we used the fura-2 method to examine whether a change in [Ca 2+ ] e alters [Ca 2+ ] i levels in bone marrow stromal cells. Changes of [Ca 2+ ] e from 1.8 mM to 5.4 mM and 10.8 mM significantly increased [Ca 2+ ] i by 1.1 and 1.3 times, respectively. Next, bone marrow stromal cells were cultured for 14 days in high [Ca 2+ ] e (5.4 mM and 10.8 mM) and normal [Ca 2+ ] e (1.8 mM) conditions. Adipocyte development was monitored by Oil Red O staining of cytoplasmic lipids and by the activity of glycerol-3-phosphate dehydrogenase (GPDH). In 5.4 mM and 10.8 mM [Ca 2+ ] e , Oil Red O-stained cells increased significantly by 1.4 and 2.3 times, respectively, and GPDH activity increased significantly by 1.7 and 2.3 times, respectively, compared with the respective values in 1.8 mM [Ca 2+ ] e . Conclusions: These results indicate that high [Ca 2+ ] e induces an increase of [Ca 2+ ] i , which enhances adipocyte development in bone marrow stroma. Further studies are required to determine the influx pathway of Ca 2+ , since prevention of Ca 2+ influx into bone marrow stromal cells might suppress development of fatty marrow and reduce age-related fracture and anemia.

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