Relationship Between Liver Biological Rhythm And Intestinal Flora In Obese Rats

[ Objective] Existing studies have shown that the occurrence of obesity will affect the expression level of biological rhythm,and will cause intestinal flora imbalance,but the relationship between obesity biorhythm and intestinal flora has rarely been reported.The relationship between the rat Clock,Per3 and Arntl clock genes,the expression level of Scd1 gene and intestinal flora,and the regulation of GQD,provide GQD prevention and treatment of high fat diet-induced obesity from a biological rhythm perspective.Clinical drug reference,for clinical reference for pre-diabetes research.[Methods] In order to simulate the eating behavior of human hyperlipidemia with obesity,this experiment induced a nutritional obesity rat model through high-fat diet,and gave high-fat diet and normal feed to the model group and the blank group respectively.End ileal histopathological sections,HE staining and WB experiments were performed.The model was further determined.The model group was given GQD intervention,and the q-PCR?TaqMan?method was used to detect the mRNA expression levels of the clock genes Clock,Arntl,Per3 and Scd1 in the liver of rats.16 S rRNA sequencing technology was used to sequence the intestinal flora of rats.Finally,the relationship between the clock genes Clock,Per3,Arntl and Scd1 and intestinal flora was analyzed based on bioinformatics.[ Results] 1.Results of animal experiment: After giving 60% high-fat diet for 1 week,the body weight and body length were significantly different compared with the blank group?P<0.01?.At the 4th week of modeling,the triglyceride level of the model group was also significantly higher.In the normal group?P<0.01?,triglyceride levels decreased significantly after 12 weeks of GOD intervention?P<0.01?.2.HE staining and WB results: After 16 weeks of modeling,the ileal mucosal epithelial cells of the blank group were closely arranged,and the defects were rare.The inherent membrane structure is normal and no cracks occur.The muscular layer of the intestinal wall is normal and tightly bound to the lamina propria.The model group has short and uneven villi,and there are many fat cells between the villi and clearly visible.Compared with the blank group fed with normal diet,the expression of claudin1,IAP and ZO1 protein in the high-fat model group was down-regulated,and the expression difference in the occluding group was not obvious.3.q-PCR results: After 12 weeks after administration,the expression of Clock and Arntl genes in the model group was higher than that in the blank group?P<0.05?,Per3 and Scd1 were lower than the blank group?P<0.05?,and the GQD model was compared.The expression levels of Clock and Arntl gene expression in liver were up-regulated?P<0.05?,and the expression levels of Per3 and Scd1 mRNA were not significantly affected?P>0.05?.4.16 S rRNA results: Compared with the blank group,the model group has the same species but different proportions.The blank group was the highest?56.32%?,followed by the posterior wall?40.32%?and the micro-bacteria?1.99%?.In the model group,the posterior wall of the model group was the highest?57.00%?,the second was the Bacteroides?33.84%?,and the second was the bacteria?7.06%?.At the genus level,the difference between the two groups was larger.The blank group norank_f<sub>Bacteroidales_S24-7_group was the highest?38.59%?,Bacteroides was second?17.03%?,Lachnospiraceae_NK4A136_group?10.31%?was the second,unclassified_f<sub>Lachnospiraceae?4.02%?;the norank_f<sub>Bacteroidales_S24-7_group was the highest in the model group?23.83%?,Bacteroides followed?9.26%?and Clostridium_sensu_stricto₁?7.65%?.After the drug intervention,simvastatin?SIM?in the positive drug group had almost no change in the intestinal flora,which was similar to the structure of the model group,and GQD group had a great influence on the intestinal flora structure.5.Clock,Per3 and Arntl Clock genes and Scd1 genes were associated with intestinal flora:There were four bacterial communities positively correlated with Clock: bacteroidales_s24-7_group,Ruminiclostridium₉,Anaerostipes,Tyzzerella,There were 3 kinds of bacteria with negative correlation: vadinBB60_group,Ruminococcus₁ and Turicibacter There are 8 species of bacteria positively related to Arntl: Roseburia,Epulopiscium,Blautia,Lachnospiraceae,cellulosilyticus,muciniphila,Peptococcaceae,Ruminiclostridium₉ There were 9 negative correlates: Clostridiales_vadinBB60_group,Turicibacter,unclassified_g<sub>Lactobacillus,Ruminococcus₁,lachnospiraceae_ucg-010,Lactobacillus_johnsonii,Roseburia,leptum_g<sub>norank,Ruminiclostridium There were 7 species with positive correlation with Per3: Clostridiales_vadinBB60_group,Ruminococcus₁,unclassified_g<sub>Lactobacillus,Turicibacter,Lactobacillus_johnsonii,Lachnospiraceae,leptum_g<sub>norank There were 5 species of bacteria with negative correlation: lachnospiraceae_ucg-010,Epulopiscium,unclassified_g<sub>Blautia,cellulosilyticus,and Peptococcaceae Three strains were positively correlated with Scd1: Lachnospiraceae_NK4A136_group,Lactobacillus_johnsonii,and vadinBB60_group There were 4 species of bacteria with negative correlation: Ruminiclostridium₉,cellulosilyticus,Ruminococcaceae,bacteroidales_s24-7_group[Conclusion] 1.The modeling speed and time of nutritionally obese rats induced by high-fat diet were fast.2.The adhesion ability of bacteria in the intestinal wall of the ileum of nutritionally obese rats was decreased,and obesity down-regulated claudin1,ZO1 and IAP contents and expressions of intestinal tight junction proteins,but had no significant changes in occluding expression.3.GQD significantly upregulated the expression of biothythm genes in liver of rats with nutritional obesity,and the effect was significant in the medium and low dose groups.4.The effect of GQD on the intestinal flora of nutrition-obese rats is different.When used at low doses,the flora structure will not be changed until a certain period of time.5.The important influences of Clock gene,Per3,Arntl and Scd1 gene on intestinal flora of nutritionally obese rats are Ruminiclostridium₉ is positively correlated with Clock and Arntl genes and negatively correlated with Scd1.VadinBB60_group is positively correlated with Arntl and Scd1 genes,negatively correlated with Clock,and negatively correlated with obesity and body weight.Ruminococcus₁ and Turicibacter are negatively correlated with Clock and Arntl genes,and positively correlated with Per3.It has been reported in literature that Ruminococcus₁ and Turicibacter are positively correlated with Clock and Arntl genes.