Sini San ameliorates lipid metabolism in hyperprolactinemia rat with liver-depression

Figures

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  Fig. 1. SNS improves the depression-like behavior of liver-depressed HPRL rats. (a) Sugar water preference test. (b) Tail suspension test. (c-d) Open field test. (e) PRL levels in rats. (f) FSH levels in rats. (g) E2 content in rats.Data are mean ± standard deviation, n = 6. Compared with the CON group, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Compared with the MODEL group, ##p < 0.01, ####p < 0.0001.

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  Fig. 2. SNS improved ovarian histomorphology in HPRL rats with liver-depression. (a) Normal group. (b) Model group. (c) BRO group. (d) LOW-SNS group. (e) MID-SNS group. (f) HIGH-SNS group. Scale bar = 500 μm. Yellow arrow represents primary follicles, red arrow represents secondary follicles, and black arrow represents mature follicles.

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  Fig. 3. Statistical analysis of serum samples. OPLS-DA score plots for the former, and OPLS-DA validation plots for the latter. (a) Total ion flow plots in positive and negative ion modes. (b) PCA plots of samples in each group in positive and negative ion modes. (c) MODEL group vs. CON group (d) BRO group vs. MODEL group.(e) LOW-SNS group vs. MODEL group. (f) MID-SNS group vs. MODEL group. (g) HIGH-SNS group vs. MODEL group.

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  Fig. 4. Screening of differential metabolites. (a) LOW-SNS group vs. MODEL group. (b) MID-SNS group vs. MODEL group. (c) HIGH-SNS group vs. MODEL group. The first row shows the volcano plot and the second row shows the PCA score plot. (d) Venn diagram of differential metabolites between the three groups a-c under conditions of VIP value ≥ 1 and p-value < 0.05. (e) Venn diagram of differential metabolites between the three groups a-c under the conditions of VIP value ≥ 1, p-value < 0.01, FC ≥ 3 or FC ≤ 0.33.

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  Fig. 5. Comparison of differences in the content of differential metabolites and their metabolic pathway analysis. (a) Clustering heat map of differential metabolites. (b) Differential metabolite relative content violin plot. 3 of the 44 differential metabolites have too long names and are replaced with codes. (c) KEGG enrichment analysis of differential metabolites. The color of the dot is the P-value size, the redder indicates the more significant enrichment.The size of the dot represents the number of different metabolites enriched.

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  Fig. 6. Network pharmacological analysis of SNS in the treatment of liver-depression HPRL. (a) Core active ingredients of SNS. (b) Venn diagram depicting the Intersection of SNS active compounds and disease targets. (c) Protein interaction network diagram. (d) Core target genes. (e) Traditional Chinese medicine ingredient-target-disease network diagram. (f) GO analysis of the core target genes, Molecular Function, Biological Process, and Cellular Component. (g) Enrichment analysis of the KEGG pathway of the core target genes.

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  Fig. 7. Molecular docking simulation. (a) Heat map of molecular docking. (b) The binding of AKT1 with quercetin. (c) The binding of TNF with quercetin. (d) The binding of IL6 with quercetin. (e) The binding of TP53 with kaempferol. (g) The binding of IL1B with beta-sitosterol.

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  Fig. 8. Serum from rats containing SNS significantly reduces the content of PRL released by MMQ cells. (a-c) Effects of different incubation times, inoculation densities, and CCK8 incubation times on the activity of MMQ cells. (d) Effects of different concentrations of drug-containing serum on the viability of MMQ cells. (e) Effects of different concentrations of bromocriptine mesylate on the viability of MMQ cells. (f) Effects of BRO on the content of PRL released by MMQ cells. (g) Effects of SNS-containing serum on the content of PRL released by MMQ cells in rats.Data are mean ± standard deviation, n = 4. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 compared with the CON group (drug concentration = 0).

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  Fig. S1. Base peak chromatogram of SNS water decoction, SNS-medicated serum, the blank serum.

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  Fig. S2. Overall flowchart of animal experiments and examination of the effect of liver-depression HPRL rat models. (a) Schematic diagram of the pharmacodynamic experiment in rats, which lasted for 10 weeks, with behavioral tests at weeks 5 and 10. (b) Serum PRL levels of rats after modeling. (c) Sugar water preference test. (d) Tail suspension test. (e-f) Open field test. Data are mean ± standard deviation, n = 6. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001, compared with the CON group.