Objective To observe the changes of intestinal flora in Wuzhishan miniature pig atherosclerosis model induced by high-fat and high-sugar diet.
Methods Twelve male Wuzhishan miniature pigs were divided into two groups according to body weight, namely the normal control (NC) group and the model (AS) group, with 6 pigs in each group. The NC group was fed with normal feed, while the AS group was fed with high-fat and high-sugar feed. After 24 weeks of continuous diet, blood was collected from the anterior vena cava to measure biochemical indicators such as glucose and lipid metabolism, and glucose tolerance test was performed at the same time. After the experiment, euthanasia was performed to observe the fat deposition in the body, and the aortic vessels were harvested for Sudan IV staining and abdominal aortic vessels for HE staining to observe lipid deposition and histopathological changes, respectively. The cecal contents were taken for 16S rDNA sequencing to analyze the changes of intestinal flora.
Results Compared with the NC group, serum total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), blood glucose (GLU), and fructosamine (FMN) bacteria in the AS model group were significantly higher. increased (P<0.05, P<0.01), body weight and body fat deposition were also significantly increased (P<0.01), and the aortic vessels in the AS model group had obvious lipid deposition and AS lesions. Sequencing analysis of intestinal flora showed that compared with the NC group, the α-diversity of the intestinal flora in the AS model group was significantly reduced (P<0.05, P<0.01). At the phylum level, Bacteroidetes and Actinobacteria were significantly decreased in AS model group (P<0.05, P<0.01), and the ratios of Proteobacteria and Firmicutes/Bacteroidetes were significantly increased (P<0.05). . At the genus level, the AS model group includes Hesyllic, Ruminococcas, Lachnosyiracea_ineeroe_sedis, Pseudobutyrio, Barnesia The relative abundance of 17 genera such as Barnesiella decreased significantly (P<0.05, P<0.01), Ramlibacter, Sphingorhabdus, Coacikacilus, Oxalobacter The relative abundance of 11 genera, including Oxalobacter, Sphingomonas, and Norosphingobium, increased significantly (P<0.05). Functional prediction analysis showed that multiple pathways such as lipopolysaccharide biosynthesis, lipopolysaccharide biosynthesis protein pathway, primary bile acid biosynthesis, and arachidonic acid metabolism may be involved in the formation of AS. Correlation analysis showed that TC, intima-media thickness, lipid deposition, fructosamine were associated with 14 genera including Oxalobacter, Sphingomonas, Sewerbacter, Clostridium, Ruminococcus.
Conclusion The AS model of Wuzhishan miniature pigs induced by high-fat and high-sugar diet has the characteristics of obesity, glucose and lipid metabolism disorder and obvious AS lesions, which may be related to intestinal flora imbalance.