[Animal modeling] - Changes of intestinal flora in atherosclerosis model of Wuzhishan miniature pigs induced by high-fat and high-sugar diet

  Objective To observe the changes of intestinal flora in atherosclerosis model of Wuzhishan mini pig induced by high-fat and high-sugar diet.

  Methods Twelve male Wuzhishan miniature pigs were divided into two groups according to their body weight: normal control (NC) group and model (AS) group, with 6 pigs in each group. NC group was fed with normal diet, AS group was fed with high fat and high sugar diet. After 24 weeks of continuous diet, blood was collected from anterior vena cava to measure biochemical indicators such as glucose and lipid metabolism, and glucose tolerance test was also conducted. After the experiment, the rats were euthanized to observe the fat deposition in the body, and the aorta was taken for Sudan IV staining and the abdominal aorta for HE staining to observe the 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 NC group, the total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), blood glucose (GLU), fructosamine (FMN) bacteria in serum of AS model group were significantly increased (P<0.05, P<0.01), and the body weight and body fat deposition were also significantly increased (P<0.01). At the same time, the aortic vessels of AS model group had significant lipid deposition and AS lesions. Sequence analysis of intestinal flora showed that compared with NC group, the intestinal flora of AS model group α- The diversity decreased significantly (P<0.05, P<0.01). At the phyla level, Bacteroides and Actinomycetes in AS model group decreased significantly (P<0.05, P<0.01), and the ratios of Proteus and Firmicum/Bacteroides increased significantly (P<0.05). At the genus level, the relative abundance of 17 genera in AS model group, including Hesylelic, Ruminococcas, Lachnosyiracea_ineeroe_sedis, Pseudobutyrio and Barnesiella, decreased significantly (P<0.05, P<0.01), Ramlibacter, Sphingohabdus The relative abundance of 11 genera, including Coacikcilus, Oxalobacter, Sphingomonas and Norospingobium, increased significantly (P<0.05). Functional prediction analysis showed that multiple pathways such as lipopolysaccharide biosynthesis, lipopolysaccharide biosynthetic protein pathway, primary bile acid biosynthesis, arachidonic acid metabolism may participate in the formation of AS. Correlation analysis showed that TC, intima-media thickness, lipid deposition, fructosamine were associated with 14 genera, including Oxalibacter, Sphingomonas, Enterobacter, Clostridium and Ruminococcus.

  Conclusion The AS model of Wuzhishan miniature pig induced by high-fat and high-sugar diet has the characteristics of obesity, disorder of glucose and lipid metabolism and obvious AS lesions, which may be related to the imbalance of intestinal flora.