Therefore, our objective was to characterize circulating EVs derived from platelets, macrophages, neutrophils, and hepatocytes in female mice fed a diet high in fat, fructose, and cholesterol (FFC) over time in comparison to male mice in order to report around the cellular sources of circulating EVs and the influence of biological sex on circulating EVs

Therefore, our objective was to characterize circulating EVs derived from platelets, macrophages, neutrophils, and hepatocytes in female mice fed a diet high in fat, fructose, and cholesterol (FFC) over time in comparison to male mice in order to report around the cellular sources of circulating EVs and the influence of biological sex on circulating EVs. neutrophil\derived EVs; and clusters of differentiation 61 (CD61) is usually a marker of platelet\derived EVs. Nonalcoholic fatty liver disease activity score (NAS) was calculated using hematoxylin and eosin\stained liver sections, and magnetic resonance imaging (MRI) was utilized for measurement of the excess fat portion and elastography. Hepatocyte\derived EVs increased in both male and female mice at 12 and 10 weeks of feeding, respectively, and remained elevated at 24 weeks in both male and female mice and at 48 weeks in male mice and 36 weeks in female mice. Macrophage\ and neutrophil\derived EVs were significantly elevated at 24 Donepezil hydrochloride weeks of dietary feeding concomitant with the histologic presence of inflammatory foci in the liver. In excess fat\, fructose\, and cholesterol\ (FFC) fed male mice, platelet\derived EVs were elevated at 12, 24, and 48 weeks, whereas in female mice, platelet derived EVs were significantly elevated at 24 weeks. Hepatocyte\, macrophage\ and neutrophil\derived EVs correlated well with the histologic NAS. Circulating cell\type\specific EVs may be a novel biomarker for NASH diagnosis and longitudinal follow up. Abbreviations+positiveALTalanine aminotransferaseAPCallophycocyaninASGR1asialoglycoprotein receptor 1CDclusters of differentiationCYP2E1cytochrome P4502E1EVextracellular vesicleFFCfat, fructose, and cholesterolLy\6G/6Clymphocyte antigen 6 complex, locus G/C1MREmagnetic resonance elastographyMRImagnetic resonance imagingNAFLDnonalcoholic fatty liver diseaseNASnonalcoholic fatty liver disease activity scoreNASHnonalcoholic steatohepatitisnsnot significantRCDrodent chow diet Nonalcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease worldwide.1 The progressive component of NAFLD, termed nonalcoholic steatohepatitis (NASH), is characterized by hepatocellular injury, ballooning, inflammation, and fibrosis. NAFLD, including NASH, is usually a complex and heterogeneous disorder in which the combinatorial and variable contribution of multiple pathogenic processes, such as lipotoxicity, microbiome, environment, epigenetics, genetic variants, and other unidentified factors, prospects to great individual variability.2 Biological sex is an additional key factor that influences disease phenotype.3, 4 The influence of Donepezil hydrochloride biological sex on NAFLD pathogenesis and on biomarkers being developed as a replacement to the platinum\standard liver biopsy remains incompletely understood. Circulating extracellular vesicles (EVs) are a relatively novel and promising blood\based biomarker being tested for diagnostic power in NAFLD.5, 6 Most cells release EVs into their extracellular milieu.6 Quantitative changes in EV figures or qualitative changes in EV cargoes in disease says may serve as disease\specific signatures and are being developed as potential blood\based biomarkers.5 It has been exhibited that total circulating EVs and hepatocyte\derived EVs are elevated in male mice with diet\induced NASH.7 In addition to hepatocytes, innate immune cells, especially macrophages and neutrophils, are implicated in Rabbit Polyclonal to SMC1 (phospho-Ser957) NASH8, 9; however, alternative cellular sources of EVs in NASH models have not been characterized. Epidemiologic data support an increase in NAFLD prevalence in women after menopause,10 yet overall, men are at greater risk for NAFLD and related fibrosis and hepatocellular carcinoma, even after adjusting for age and metabolic comorbidities. 11 Dietary murine models of NAFLD frequently demonstrate a similar Donepezil hydrochloride sexual dimorphism.3 Male mice from your C57BL/6J strain develop significant weight gain and associated metabolic features, such as insulin resistance and hepatic steatosis, while female mice from your same strain are relatively resistant to the effects of diet\induced obesity.12 Similarly, biomarkers of liver disease may demonstrate sexual dimorphism.13 For example, plasma alanine aminotransferase (ALT) levels and microRNA\122 levels are higher in men than women.13, 14 The influence of biological sex on circulating EVs in NAFLD has not been carefully examined. The diverse cellular sources of EVs have also not been delineated. Therefore, our objective was to characterize circulating EVs derived from platelets, macrophages, neutrophils, and hepatocytes in female mice fed a diet high in excess fat, fructose, and cholesterol (FFC) over time Donepezil hydrochloride in comparison to male mice in order to report around the cellular sources of circulating EVs and the influence of biological sex on circulating EVs. We further Donepezil hydrochloride compared circulating EVs with a histologic assessment of the well\established platinum\standard NAFLD activity score (NAS) and magnetic resonance elastography (MRE) parameters, an emerging noninvasive biomarker. Here, we demonstrate that hepatocyte\derived EVs are elevated early in NAFLD, well before histologically apparent inflammation, and remain elevated over time; furthermore, hepatocyte\derived EVs correlate well with histologic features of NASH. Materials and Methods Animal Model All animal use was approved by the institutional.