Glucose Restriction Plus Refeeding in Vitro Induce Changes of the Human Adipocyte Secretome with an Impact on Complement Factors and Cathepsins.

Adipose tissue is a major endocrine organ capable of secreting adipokines with a role in whole-body metabolism. Changes in the secretome profile during the development of obesity is suspected to contribute to the risk of health complications such as those associated with weight regain after weight loss. However, the number of studies on weight regain is limited and secretome changes during weight regain have hardly been investigated. In an attempt to generate leads for in vivo studies, we have subjected human Simpson Golabi Behmel Syndrome adipocytes to glucose restriction (GR) followed by refeeding (RF) as an in vitro surrogate for weight regain after weight loss. Using LC-MS/MS, we compared the secreted protein profile after GR plus RF with that of normal feeding (NF) to assess the consequences of GR plus RF. We identified 338 secreted proteins of which 49 were described for the first time as being secreted by adipocytes. In addition, comparison between NF and GR plus RF showed 39 differentially secreted proteins. Functional classification revealed GR plus RF-induced changes of enzymes for extracellular matrix modification, complement system factors, cathepsins, and several proteins related to Alzheimer's disease. These observations can be used as clues to investigate metabolic consequences of weight regain, weight cycling or intermittent fasting.

Salmonella-driven intestinal edema in mice is characterized by tensed fibronectin fibers.

Intestinal edema is a common manifestation of numerous gastrointestinal diseases and is characterized by the accumulation of fluid in the interstitial space of the intestinal wall. Technical advances in laser capture microdissection and low-biomass proteomics now allow us to specifically characterize the intestinal edema proteome. Using advanced proteomics, we identify peptides derived from antimicrobial factors with high signal intensity, but also highlight major contributions from the blood clotting system, extracellular matrix (ECM) and protease-protease inhibitor networks. The ECM is a complex fibrillar network of macromolecules that provides structural and mechanical support to the intestinal tissue. One abundant component of the ECM observed in Salmonella-driven intestinal edema is the glycoprotein fibronectin, recognized for its structure-function interplay regulated by mechanical forces. Using mechanosensitive staining of fibronectin fibers reveals that they are tensed in the edema, despite the high abundance of proteases able to cleave fibronectin. In contrast, fibronectin fibers increasingly relax in other cecal tissue areas as the infection progresses. Co-staining for fibrin(ogen) indicates the formation of a provisional matrix in the edema, similar to what is observed in response to skin injury, while collagen staining reveals a sparse and disrupted collagen fiber network. These observations plus the absence of low tensional fibronectin fibers and the additional finding of a high number of protease inhibitors in the edema proteome could indicate a critical role of stretched fibronectin fibers in maintaining tissue integrity in the severely inflamed cecum. Understanding these processes may also provide valuable functional diagnostic markers of intestinal disease progression in the future.

T1 mapping and feature tracking imaging of left ventricular extracellular remodeling in severe aortic stenosis.

BACKGROUND: Left ventricular (LV) extracellular remodeling is a critical process in aortic stenosis (AS), which is related to functional abnormalities. Data regarding the use of combined T1 mapping and feature tracking (FT) to assess LV extracellular remodeling in severe AS are scarce. This study aimed to investigate the ability of T1-derived and FT-derived parameters to identify and assess the changes in process of LV extracellular remodeling in patients with severe AS. METHODS: A total of 49 patients with severe AS and 20 healthy volunteers were prospectively recruited. Modified look-locker inversion-recovery T1 mapping and FT imaging were performed in all participants using 3.0-T cardiac magnetic resonance imaging. The degree of myocardial fibrosis was quantified using Masson trichrome stain in biopsy specimens obtained intraoperatively from 13 patients and expressed as collagen volume fraction (CVF). Patients were divided into subgroups according to preserved LV ejection fraction (LVEF) (LVEF ≥50%) or reduced LVEF (LVEF <50%). RESULTS: Regarding the diffuse fibrosis burden, extracellular volume (ECV) was statistically insignificant between patients with preserved LVEF) and controls (28.0%±3.3% vs. 26.5%±2.3%, P>0.05). ECV in the reduced LVEF group (n=20) was significantly higher than that in the preserved LVEF group (n=29) (30.4%±3.9% vs. 28.0%±3.3%, P<0.05). Regarding the myocardial strain, global longitudinal strain (GLS) showed increasing impairment from the control group to the preserved LVEF AS group to the reduced LVEF AS group (-23.4%±3.3% vs. -18.6%±3.8% vs. -11.2%±4.8%, P<0.05). A significant correlation was found between ECV and CVF (r=0.64, P=0.020), whereas the correlation between GLS and CVF was insignificant. Significant correlations were observed between GLS and LV mass index (r=0.72, P=0.006) and LVEF (r=0.82, P<0.001). However, no correlations were found between ECV and LV mass index (P=0.172) and between ECV and LVEF (P=0.339). Discrimination of patients with preserved LVEF from controls, GLS yielded the best diagnostic performance as defined by the area of under the curve (-0.83), and GLS, ECV, and post-T1 were significant discriminators after regression analysis. CONCLUSIONS: In the process of LV extracellular remodeling in severe AS, ECV is the structural marker of extracellular fibrosis burden, and GLS is the functional marker before the fibrosis burden intensifies.

Saroglitazar reduces obesity and associated inflammatory consequences in murine adipose tissue.

Prevailing knowledge links chronic low-grade inflammation in the adipose tissue to obesity and its associated metabolic complications. In this study, we evaluated immunometabolic effects of a recently launched dual peroxisome proliferator-activated receptor (PPAR) α & γ agonist 'Saroglitazar' in a mouse model of diet-induced obesity (DIO). Body composition analysis revealed that saroglitazar treatment promoted hepatic weight gain, while attenuated epididymal white adipose tissue (eWAT) mass in DIO. In the eWAT of saroglitazar treated mice, histological analysis showed reduced adipocyte hypertrophy and matrix deposition (picrosirius red staining). Immunological profiling of stromal vascular fraction isolated from eWAT showed decreased pro-inflammatory cells (M1 macrophages, CD4 and CD8 T-cells) and increased anti-inflammatory M2 macrophages. Gene expression and western blot analysis suggested that saroglitazar promoted energy expenditure machinery and attenuated inflammatory as well as fibrotic markers in eWAT during DIO. In conclusion, for the first time we are reporting immunometabolic effects of dual PPARα & γ agonist saroglitazar in DIO and insulin resistance (IR). Saroglitazar exerted its beneficial effects on adipose tissue by limiting, diet-induced adipose tissue dysfunction, adipocyte hypertrophy, adipocyte cell damage and extracellular matrix deposition in obesity.