Comprehensive multiomics analysis of the effect of ginsenoside Rb1 on hyperlipidemia

Figure 1. Characteristics of 16S rRNA sequencing. (A) TaxonTree revealed the distribution of each species in all samples. Each layer in the leaf node represents a classification level, from left to right is the boundary, gate, class, order, family, genus and species level, only genus annotated and with relative abundance large than 1% was shown in the figure. (B) PCA of OUT between Control and Model group. Each point in the graph represents a sample. The different colors of the points represent the grouping of the samples. (C) PCA of OUT between Rb1 and Model group. (D) Relative abundance of bacteria in genus level in each group. Blautia and Allobaculum increased in Model group and Turicibacter decreased in Model group. Notes: data was shown in mean +/- SD, * represent P<0.05, ** represent P<0.01, *** represent P<0.001. (E) KEGG analysis of abundance difference between Rb1 and model group. The result was performed by PICRUSt software with default parameters. Each color represents a group of samples, their average relative abundance is showed in X axis. The left of bar chart shows the pathways with significant differences in abundance between the two groups, and the right of chart shows P…

Figure 2. Characteristics of lipidomics in positive ion model. (A) Score scatter plot of OPLS-DA model for group C vs M in POS model. The X axis t [1] P denotes the predicted principal component score of the first principal component, the Y axis t [1] O denotes the orthogonal principal component scores. The two groups of samples are very distinct in this model. C, control group. M, model group. POS, positive ion model. (B) Score scatter plot of OPLS-DA model for group M vs R in POS. M, model group. R, Rb1 group. POS, positive ion model (C) Permutation test of OPLS-DA model for group Control vs Model in POS. The X axis represents the replacement retention, the Y axis represents the value of R2Y or Q2, the green dot represents the value of R2Y and the blue square represents the value of Q of the replacement test. The dotted lines represent the regression lines of R Y and Q, respectively. The original model can well explain the difference between the two groups of samples. (D) Permutation test of OPLS-DA model for group Rb1 vs Model in POS. (E) Pathway analysis for group Control vs Model in POS. The bubble size indicates the influencing factor in the topological analysis; the bubble color…

Figure 3. Characteristics of transcriptomics. (A) Heat map of intersample correlation between model and control group. The correlation was evaluated by Pearson’s correlation coefficient, different color represents differences among different samples. (B) Heat map of intersample correlation between Rb1 and model group. (C) FPKM distribution boxes of transcripts in each sample. Y axis is shown in log10 scale, X axis one sample per region. (D) Clustering map of differential genes. The depth of the color represents the level of gene expression. (E) KEGG pathway analysis of differentially expressed genes in control group versus model group. Gene ratio represents enrichment factor, bubble scale represents number of different genes, depth of bubble color represents P value. (F) KEGG pathway analysis of differentially expressed genes in Rb1 group versus model group.

Figure 4. Comprehensive analysis of microbiology, lipidomics and transcriptomics. (A) Lipidomics showed content of PC increase in normal mice of Rb1 group comparing with that of Model group. Y axis is shown in foldchange, X axis shows different electron cut-off points of PC. All the metabolites with VIP >1 and P <0.05.PC, phosphatidylcholine. POS, positive ion model. NEG, negative ion model. (B) RNA-seq and qRT-PCR showed expression of RNAs associated with glycerophospholipid metabolism. RNA-seq was performed once, PCR were repeated thrice, n=3, data was shown in mean +/- SD. (C) Serum level of hyperlipidemia associated index showed Rb1 reduced total cholesterol level. TG, triglycerides; TC: total cholesterol; LDL, low density lipoprotein; HDL, high density lipoprotein;** represents P<0.01, n=6. (D) Lipidomics showed content of PC increase in fecal transplanted germ-free mice of Rb1 group comparing with that of Model group. Y axis is shown in foldchange, X axis shows different electron cut-off points of PC. All the metabolites with VIP >1 and P <0.05.PC, phosphatidylcholine. (E) QRT-PCR showed that Pmet increased and Dgkg, Phospho1, Pla4g4b decreased in fecal transplanted germ-free mice of…

Figure 5. RB1 increase phosphatidylcholine in glycerophospholipid metabolism to prevent hyperlipidemia. Through multi-omics analysis, we found Glycerophospholipid metabolism was a common pathway between Rb1 and model group. By using germ-free mice, we found Rb1 increased the abundance of Turicibacter and decreased the abundance of Blautia and Allobaculum to increase content of phosphatidylcholine by raising Pemt and reducing Pla2g4b and Phospho1, and resulting the downregulation of total cholesterol to play a prevention role in hyperlipideamia. Notes: The green box in mocrobe and metabolism box represents the content reduced, the orange box represents the content increased. Pemt promotes synthesis of phosphatidylcholine, Pla2g4b and Phospho1 accelerate decomposition of phosphatidylcholine. Notes: Experiments were repeated twice, data was shown in mean +/- SD, * represent P<0.05, ** represent P<0.01, *** represent P<0.001.