Diet-induced changes in mouse cells in vitro and in vivo zebrafish models of angiogenesis

Abstract

Cardiovascular disorders rank as the primary cause of global mortality. Being overweight or obese impacts the pathogenesis of cardiovascular disease, resulting in an imbalance in endothelial function, cell growth, and inflammatory activation. Disruption of these factors resulting from endothelial cell dysfunction serves as both an outcome and a catalyst for vascular disease processes. Endothelial cells (ECs) are a natural barrier between circulating blood and vessel components. They also play critical roles in multiple physiological and pathophysiological processes, such as angiogenesis, vascular permeability, and inflammation. Amelioration of endothelial dysfunction may be attained by weight loss; however, complementary in vitro and in vivo studies are needed to establish the effects of weight loss on endothelial function and angiogenesis. This study developed an in vitro model to understand better the diet-induced changes in angiogenesis for mouse endothelial cells. In addition, a novel in vivo model of diet-induced vascular changes and its potential reversal with a return to regular diet in a zebrafish model was also studied. In vitro studies showed that a serum from mice fed a high-fat diet (HFD) might lead to proliferation of endothelial cells, yet weight loss did not compensate for prior stress induced by HFD. In vivo, studies in adult zebrafish showed that egg yolk-based high-fat diet might affect cytological architecture in the adult fish liver. Switching to a normal diet could effectively reverse these changes. Moreover, a caudal fin inter-ray vascularization assay was developed and used to test whether vessel sprouting was affected by different diets. Overfeeding resulted in a higher number of vessels, yet future studies with higher sample sizes are needed. Similarly, the expressions of several angiogenesis-related genes, which were quantified using cDNAs from the whole larvae and adult caudal fin treated with different diets, showed significant changes in vcam in larvae and cdh5 in adult fin by diet. However, further experiments are needed due to high individual variability and low sample size. The findings herein show that in vitro mouse endothelial cells and zebrafish larvae and adults could be used as valuable models for studies involving reversal/weight loss of high fat or overfeeding dietary regimes. Furthermore, the caudal fin vascularization assay in Tg(fli1:eGFP) Casper fish could be a promising preclinical model for testing the effects of different diets on angiogenesis and endothelial dysfunction.