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Accumulating evidence indicates that lactic acid bacteria could improve host physiology and lipid metabolism. To investigate the effect of the gut microbiota on host lipid metabolism, a hyperlipidemic rat model was established by feeding rats a high-fat diet for 28 days, and the gut microbiota of the rats was analyzed using real-time PCR before and after administration of Lactobacillus rhamnosus hsryfm 1301 and its fermented milk for 28 days. The findings showed that the Lactobacillus spp., Bifidobacterium spp., Bacteroides spp., and Enterococcus spp. content in the hyperlipidemic rats gut was increased significantly (p < 0.05), while the Clostridium leptum and Enterobacter spp. content was decreased significantly after intervening with L. rhamnosus hrsyfm 1301 and its fermented milk for 28 days (p < 0.05). Furthermore, the lipid levels of the serum and the liver were decreased significantly (p < 0.05) and the fecal water content was increased significantly (p < 0.05) in the hyperlipidemic rats after the intervention, and hepatocyte fatty degeneration of liver tissues was also prevented. A positive correlation was observed between the Clostridium leptum content and the level of serum cholesterol, triglycerides, low-density lipoprotein, and high-density lipoprotein, and a negative correlation was observed between the Enterobacter spp. content and the Lactobacillus spp. and Bifidobacterium spp. content in the hyperlipidemic rats gut. These results suggest that the gut microbiota and lipid metabolism of hyperlipidemic rats could be improved by supplementation with L. rhamnosus hsryfm 1301 and its fermented milk.

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