Abstract P067: Piezo1 Activation Inhibits Adipogenesis And Lipogenesis In Perivascular Adipose Tissue Preadipocytes

Hypertension ◽  
2020 ◽  
Vol 76 (Suppl_1) ◽  
Author(s):  
Cristian Javier Rendon Mora ◽  
Emma D Flood ◽  
Stephanie W Watts ◽  
G.Andres Contreras ◽  
Janice Thompson
Keyword(s):  
Adipose Tissue ◽  
Expression Patterns ◽  
Stromal Vascular Fraction ◽  
Serial Passage ◽  
Cell System ◽  
Sprague Dawley ◽  
Perivascular Adipose Tissue ◽  
Triglyceride Accumulation ◽  
Sprague Dawley Rats ◽  
Triglyceride Content

In adipogenesis, perivascular adipose tissue (PVAT) preadipocytes turn into adipocytes. In non-PVAT preadipocytes, mechanical forces affect the commitment and lipogenic stages of adipogenesis. The mechanism may involve PIEZO1, a mechanosensor, that boosts differentiation of progenitor cells towards osteogenic and fibroblastic lineages. Since hypertension causes changes in the vascular forces that could affect adipogenesis in PVAT, our objective was to evaluate PIEZO1 expression patterns in PVAT and the effects of PIEZO1 activation on the adipogenic potential of preadipocytes. We hypothesize that PIEZO1 activation limits the adipogenic potential of PVAT preadipocytes. PVAT from the thoracic aorta (APVAT) was collected from male Sprague Dawley rats at 10 weeks of age (n=5). Preadipocytes were obtained by Liberase digestion followed by serial passage of adherent cells. Preadipocyte progenitors, CD34+PDGFRα+, were harvested by magnetic-activated sorting. PIEZO1 expression was assessed by RT-qPCR and immunofluorescence (IF). Preadipocytes were induced to differentiate for 14 d in standard media (CON) or in the presence of PIEZO1 agonist Yoda (10μM) and inhibitor Dooku (5μM) during days 0-2 (commitment), 3-14 (lipogenesis), and 0-14 (overall adipogenesis). Adipogenesis was evaluated with IncuCyte Live-Cell system using the triglyceride stain Bodipy 493503. Triglyceride content is reported as Bodipy Intensity/Adipocyte Count. Piezo1 RNA was expressed in adipocytes and the stromal vascular fraction of APVAT. PIEZO1 IF signal was detected in SVF and preadipocyte. Triglyceride was reduced by Yoda (62 ± 14.3) and Dooku (49.3 ± 14) during 0-2 d compared to CON (312.5 ± 165.6). Neither Yoda nor Dooku for 12 d affected triglyceride accumulation compared to CON. In contrast, the lipid content of Yoda (77.7 ± 21.3) and Dooku (48.9 ± 15.2) treated cells during 14 d was reduced vs CON (312.5 ± 165.6). The expression of PIEZO1 in all PVAT fractions suggests mechanosensitivity. PIEZO1 activation during adipogenesis commitment impaired adipocyte maturation. These data provide evidence for the capacity of mechanosensory expressed in PVAT preadipocytes to modulate adipogenesis, underpinning the deleterious impact of hypertension on PVAT function.

scholarly journals Effect of Resistance Training on NADPH Oxidase and Adiponectin in PVAT of OVX Rats

2021 ◽  
Vol 30 (2) ◽  
pp. 205-212
Author(s):  
Erling Guo ◽  
Jin-Hwan Yoon ◽  
Wooyeon Jo ◽  
Jaeho Jin ◽  
Sang Ki Lee
Keyword(s):  
Adipose Tissue ◽  
Resistance Training ◽  
Nadph Oxidase ◽  
Abdominal Aorta ◽  
Sham Group ◽  
Ovariectomized Rats ◽  
Sprague Dawley ◽  
Perivascular Adipose Tissue ◽  
Ovx Rats ◽  
Sprague Dawley Rats

PURPOSE: Perivascular adipose tissue (PVAT) is a type of adipose tissue that surrounds vessels to provide anti-contractile effects. This study aimed to investigate the effect of resistance training on NADPH oxidase, adiponectin, and endothelial NOS (eNOS) expression in the abdominal aorta and PVAT of ovariectomized rats.METHODS: Sprague-Dawley rats at 20 weeks of age were divided into three groups: sham control (Sham, n=10), OVX-control (OVX_ Con, n=10), and OVX-resistance exercise (OVX_Rex, n=10). Resistance training was performed by climbing a ladder for 12 weeks. Western blotting was used to analyze target protein expression in the rat abdominal aorta and PVAT.RESULTS: NADPH oxidase (p67phox) expression was significantly higher in the OVX_Con group than in the sham group, but it was significantly decreased in the OVX_Rex group. The expression of adiponectin, AKT, and eNOS in both abdominal aorta and PVAT was significantly reduced in the OVX_Con group than in the Sham group, but it was improved in the OVX_Rex group.CONCLUSIONS:The results suggest that regular resistance training inhibits p67phox and increases adiponectin expression and phosphorylation of AKT and eNOS in abdominal aortic PVAT of ovariectomized rats.

scholarly journals Loss of anti-contractile effect of perivascular adipose tissue in offspring of obese rats

2016 ◽  
Vol 40 (8) ◽  
pp. 1205-1214 ◽  
Author(s):  
K E Zaborska ◽  
M Wareing ◽  
G Edwards ◽  
C Austin
Keyword(s):  
Adipose Tissue ◽  
Maternal Obesity ◽  
Later Life ◽  
Independent Effect ◽  
Sprague Dawley ◽  
Response Curves ◽  
Perivascular Adipose Tissue ◽  
Contractile Effect ◽  
Sprague Dawley Rats ◽  
Pregnancy And Lactation

Abstract Rationale: Maternal obesity pre-programmes offspring to develop obesity and associated cardiovascular disease. Perivascular adipose tissue (PVAT) exerts an anti-contractile effect on the vasculature, which is reduced in hypertension and obesity. Objective: The objective of this study was to determine whether maternal obesity pre-programmes offspring to develop PVAT dysfunction in later life. Methods: Female Sprague–Dawley rats were fed a diet containing 10% (control) or 45% fat (high fat diet, HFD) for 12 weeks prior to mating and during pregnancy and lactation. Male offspring were killed at 12 or 24 weeks of age and tension in PVAT-intact or -denuded mesenteric artery segments was measured isometrically. Concentration–response curves were constructed to U46619 and norepinephrine. Results: Only 24-week-old HFD offspring were hypertensive (P<0.0001), although the anti-contractile effect of PVAT was lost in vessels from HFD offspring of each age. Inhibition of nitric oxide (NO) synthase with 100 μM l-NMMA attenuated the anti-contractile effect of PVAT and increased contractility of PVAT-denuded arteries (P<0.05, P<0.0001). The increase in contraction was smaller in PVAT-intact than PVAT-denuded vessels from 12-week-old HFD offspring, suggesting decreased PVAT-derived NO and release of a contractile factor (P<0.07). An additional, NO-independent effect of PVAT was evident only in norepinephrine-contracted vessels. Activation of AMP-activated kinase (with 10 μM A769662) was anti-contractile in PVAT-denuded (P<0.0001) and -intact (P<0.01) vessels and was due solely to NO in controls; the AMPK effect was similar in HFD offspring vessels (P<0.001 and P<0.01, respectively) but was partially NO-independent. Conclusions: The diminished anti-contractile effects of PVAT in offspring of HFD dams are primarily due to release of a PVAT-derived contractile factor and reduced NO bioavailability.

scholarly journals Transglutaminases Are Active in Perivascular Adipose Tissue

2021 ◽  
Vol 22 (5) ◽  
pp. 2649
Author(s):  
Alexis N. Orr ◽  
Janice M. Thompson ◽  
Janae M. Lyttle ◽  
Stephanie W. Watts
Keyword(s):  
Adipose Tissue ◽  
Vascular Remodeling ◽  
Coagulation Factor ◽  
Sprague Dawley ◽  
Rt Pcr ◽  
Ideal Model ◽  
Perivascular Adipose Tissue ◽  
Tissue Sections ◽  
Insight Into ◽  
Immunohistochemical Analyses

Transglutaminases (TGs) are crosslinking enzymes best known for their vascular remodeling in hypertension. They require calcium to form an isopeptide bond, connecting a glutamine to a protein bound lysine residue or a free amine donor such as norepinephrine (NE) or serotonin (5-HT). We discovered that perivascular adipose tissue (PVAT) contains significant amounts of these amines, making PVAT an ideal model to test interactions of amines and TGs. We hypothesized that transglutaminases are active in PVAT. Real time RT-PCR determined that Sprague Dawley rat aortic, superior mesenteric artery (SMA), and mesenteric resistance vessel (MR) PVATs express TG2 and blood coagulation Factor-XIII (FXIII) mRNA. Consistent with this, immunohistochemical analyses support that these PVATs all express TG2 and FXIII protein. The activity of TG2 and FXIII was investigated in tissue sections using substrate peptides that label active TGs when in a catalyzing calcium solution. Both TG2 and FXIII were active in rat aortic PVAT, SMAPVAT, and MRPVAT. Western blot analysis determined that the known TG inhibitor cystamine reduced incorporation of experimentally added amine donor 5-(biotinamido)pentylamine (BAP) into MRPVAT. Finally, experimentally added NE competitively inhibited incorporation of BAP into MRPVAT adipocytes. Further studies to determine the identity of amidated proteins will give insight into how these enzymes contribute to functions of PVAT and, ultimately, blood pressure.

Sepsis induces changes in the expression and distribution of Toll-like receptor 4 in the rat kidney

2006 ◽  
Vol 290 (5) ◽  
pp. F1034-F1043 ◽  
Author(s):  
Tarek M. El-Achkar ◽  
Xiaoping Huang ◽  
Zoya Plotkin ◽  
Ruben M. Sandoval ◽  
Georges J. Rhodes ◽  
...  
Keyword(s):  
Rat Kidney ◽  
Expression Patterns ◽  
Cecal Ligation ◽  
Proximal Tubules ◽  
Microbial Pathogens ◽  
Sprague Dawley ◽  
Renal Vasculature ◽  
Two Photon Microscopy ◽  
Sprague Dawley Rats ◽  
Cellular Signal

Toll-like receptors (TLRs) are now recognized as the major receptors for microbial pathogens on cells of the innate immune system. Recently, TLRs were also identified in many organs including the kidney. However, the cellular distribution and role of these renal TLRs remain largely unknown. In this paper, we investigated the expression of TLR4 in a cecal ligation and puncture (CLP) model of sepsis in Sprague-Dawley rats utilizing fluorescence microscopy. In sham animals, TLR4 was expressed predominantly in Tamm-Horsfall protein (THP)-positive tubules. In CLP animals, TLR4 expression increased markedly in all tubules (proximal and distal), glomeruli, and the renal vasculature. The staining showed a strong apical distribution in all tubules. A moderately less intense cellular signal colocalized partially with the Golgi apparatus. In addition, kidneys from septic rats showed increased expression of CD14 and THP. They each colocalized strongly with TLR4, albeit in different tubular segments. We also imaged the kidneys of live septic animals with two-photon microscopy after fluorescent lipopolysaccharide (LPS) injection. Within 10 min, LPS was seen at the brush border of some proximal tubules. Within 60 min, LPS was fully cytoplasmic in proximal tubules. Conversely, distal tubules showed no LPS uptake. We conclude that TLR4, CD14, and THP have specific renal cellular and tubular expression patterns that are markedly affected by sepsis. Systemic endotoxin can freely access the tubular and cellular sites where these proteins are present. Therefore, locally expressed TLRs and other interacting proteins could potentially modulate the renal response to systemic sepsis.

scholarly journals Antiobesity Activity of Elateriospermum tapos Shell Extract in Obesity-Induced Sprague Dawley Rats

Molecules ◽  
2021 ◽  
Vol 26 (2) ◽  
pp. 321
Author(s):  
Kokila Vani Perumal ◽  
Nor Liyana Ja’afar ◽  
Che Norma Mat Taib ◽  
Nurul Husna Shafie ◽  
Hasnah Bahari
Keyword(s):  
Body Weight ◽  
Corn Starch ◽  
Milk Powder ◽  
Caloric Intake ◽  
Lipid Lowering ◽  
Sprague Dawley ◽  
Triglyceride Accumulation ◽  
Sprague Dawley Rats ◽  
Effective Result ◽  
Lipid Lowering Effect

Obesity is one of the risk factors associated with cardiovascular diseases, hypertension, abnormal liver function, diabetes, and cancers. Orlistat is currently available to treat obesity, but it is associated with adverse side effects. Natural resources are widely used for obesity treatment. Hence, this study aimed to investigate the anti-obesity activity of Elateriospermum tapos (E. tapos) shell extract in obesity induced Sprague Dawley rats. The rats’ obesity was induced by a high-fat (HF) diet made up of 50% standard rat pellet, 20% milk powder, 6% corn starch, and 24% ghee and a cafeteria (CAF) diet such as chicken rolls, salty biscuits, cakes, and cheese snacks. A hot aqueous method for the extraction of E. tapos shells was applied by using 500 mL of distilled water for about 24 h. Various dosages of E. tapos shell extract (10 mg/kg, 100 mg/kg, and 200 mg/kg) were used. At the end of the study, body weight, caloric intake, organ weight, lipid profile, lipoprotein lipase (LPL) activity, and histopathology analysis were carried out. E. tapos shell extract treated groups showed a reduction in body weight, positive lipid-lowering effect, decrements in triglyceride accumulation and LPL activity, and positive improvement in histopathology analysis. A dose of 200 mg/kg showed the most effective result compared to 10 mg/kg and 100 mg/kg doses.

134 The Effect of Adipose Derived Stromal Vascular Fraction on Stasis Zone in an Experimental Burn Model on Streptozocin – Induced Diabetic Rats

2021 ◽  
Vol 42 (Supplement_1) ◽  
pp. S89-S90
Author(s):  
Cagri A Uysal ◽  
Burak Ozkan ◽  
Abbas Najimaldin Muhsun Al Bayati ◽  
Gonca Ozgun ◽  
Kadri Akinci ◽  
...  
Keyword(s):  
Blood Glucose ◽  
Cell Density ◽  
Inflammatory Cell ◽  
Burn Injury ◽  
Stromal Vascular Fraction ◽  
Vascular Density ◽  
Sprague Dawley ◽  
Glucose Levels ◽  
Blood Glucose Levels ◽  
Sprague Dawley Rats

Abstract Introduction Stasis zone is the encircling area of the coagulation zone which is a critical area determining the depth and width of the necrosis in burn patients. In our study we aim to salvage the stasis zone by injecting adipose derived stromal vascular fraction (ADSVF). Methods Intraperitoneal Streptozotocin was administered for the induction of diabetes mellitus (DM) and the development of DM was confirmed by the measurement of blood glucose levels in the blood samples with blood glucometer weekly 48 hours after injection. Rats with blood glucose levels above 200 mg/dl were accepted as diabetic. The diabetic animals were followed for 4 weeks before the intervention. Thermal injury was applied on dorsum of diabetic Sprague – Dawley rats (n=20) according to the previously described ‘‘comb burn’’ model. After the burn injury (30 minutes) on Sprague - Dawley rats; rat dorsum was separated into 2 equal parts consisting of 4 burn zones (3 stasis zone) on each pair. ADSVF cells harvested from inguinal fat pads of diabetic Sprague - Dawley rats (n=5) were injected on the right side while same amount of phosphate buffered saline (PBS) injected on the left side of the same animal. One week later, average vital tissue on the statis zone was determined by macroscopy, angiography and microscopy. Vascular density, inflammatory cell density and gradient of fibrosis were determined via immunohistochemical assay. Results Macroscopic stasis zone tissue survivability percentage (32 ± 3.28 %, 57 ± 4.28 %), average number of vessels (10.28 ± 1.28, 19.43 ± 1.72), capillary count (15.67 ± 1.97, 25.35 ± 2.15) and vascular density (1.55 ± 0.38, 2.14 ± 0.45) were higher on ADSVF side. Fibrosis gradient (1.87 ± 0.51, 1.50 ± 0.43) and inflammatory cell density (1.33 ± 0.40, 1.20 ± 0.32) were higher on the PBS side. Conclusions Macroscopic and microscopic findings determined that ADSVF has a statistically significant benefit for salvaging stasis zone on acute burn injuries in DM.

Regression of white adipose tissue in diabetic rats

1989 ◽  
Vol 257 (4) ◽  
pp. E547-E553 ◽  
Author(s):  
A. Geloen ◽  
P. E. Roy ◽  
L. J. Bukowiecki
Keyword(s):  
Endothelial Cells ◽  
Adipose Tissue ◽  
Protein Content ◽  
Cell Number ◽  
Diabetic Rats ◽  
Total Protein Content ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Wet Weight

The effects of long-term diabetes (4 wk) on the development of parametrial (PWAT) and retroperitoneal (RWAT) white adipose tissues were studied in young Sprague-Dawley rats (170-200 g) injected with a single dose of streptozotocin (75 mg/kg). Diabetes stopped animal growth and totally abolished the normal increases in the wet weight, total protein content, and cellularity (estimated by DNA content) of PWAT and RWAT. Remarkably, the prolonged lack of insulin induced a progressive decrease of the cellularity of RWAT to levels that were lower than those of the initial controls. It also resulted in a marked reduction of adipocyte size. The tiny adipocytes seen in diabetic animals were characterized by the presence of multilocular triglyceride droplets. In general, the decreases in cell number, cell size, and protein content were more pronounced in RWAT than in PWAT. Quantitative cellular frequency studies revealed that adipocytes, and possibly also endothelial cells, contribute to the decrease in RWAT cellularity. The results demonstrate that 1) diabetes inhibits proliferative activity in adipose tissue, 2) total cell number reduction may occur in adipose depot of young growing rats, 3) this effect is depot dependent, and 4) the turnover of adipocytes and endothelial cells is relatively slow (several weeks).

Weight reduction increases adipose but decreases cardiac LPL in reduced-obese Zucker rats

1991 ◽  
Vol 261 (2) ◽  
pp. E246-E251 ◽  
Author(s):  
D. H. Bessesen ◽  
A. D. Robertson ◽  
R. H. Eckel
Keyword(s):  
Adipose Tissue ◽  
Cardiac Muscle ◽  
Weight Reduction ◽  
Zucker Rats ◽  
Mrna Levels ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Obese Rats ◽  
Obese Zucker Rats ◽  
Coordinate Changes

Lipoprotein lipase (LPL) activity and mRNA levels were measured in cardiac muscle and adipose tissue from lean, obese, and weight-stable reduced-obese Zucker rats, both fasted and 2 h after feeding. Fasting epididymal fat LPL activity was substantially higher in obese rats relative to lean rats [6.9 vs. 0.2 nmol free fatty acid (FFA).10(6) cells-1.min-1; P = 0.0001], and was higher still in reduced-obese rats (15.7 nmol FFA.10(6) cells-1.min-1; P = 0.002). Adipose tissue LPL increased with feeding in all three groups. In marked contrast, fasting cardiac muscle LPL was lower in obese rats relative to lean (28.8 vs. 38.5 nmol FFA.g-1.min-1; P = 0.0064) and was lower still in reduced-obese rats (14.5 nmol FFA.g-1.min-1; P = 0.0001). LPL mRNA levels increased in adipose tissue along with enzyme activity; however, the magnitude of the changes were relatively small, suggesting that the primary regulatory steps are posttranslational. Weight reduction studies were also carried out in Sprague-Dawley rats with similar results. These studies show that sustained weight reduction results in coordinate changes in tissue-specific LPL, favoring delivery of lipoprotein triglyceride fatty acids to adipose tissue relative to cardiac muscle and the restoration of energy stores.

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