Multi-omics analysis reveals the pathogenesis of db/db mice diabetic kidney disease and the treatment mechanisms of multi-bioactive compounds combination from Salvia miltiorrhiza

FIGURE 1. (A). Multi-week weight change curve (n = 10); (B). Blood glucose change curve (n = 10); (C). Determination of biochemical indicators among control group, model group and administration groups; (# p < 0.05; ## p < 0.01; ### p < 0.001: models vs. control; * p < 0.05; ** p < 0.01; *** p < 0.001: treatment groups vs. models).

FIGURE 2. (A). Pathological section of kidney tissue revealed by HE staining (×200); (B). Pathological section score; (C). Pathological section of kidney tissue revealed by PAS staining (×400); (D). Average optical (# p < 0.05; ## p < 0.01; ### p < 0.001: models vs control; * p < 0.05; ** p < 0.01; *** p < 0.001: treatment groups vs models) (Black arrow: the cell nucleus was deep-stained or fragmented and dissolved. Red arrow: necrosis, structural disorder in the glomerular cells. Green arrow: a small amount of kidney tubular necrosis calcification. Yellow arrow: inflammatory cells in the tubulointerstitial).

FIGURE 3. (A). Summary of pathway analysis with MetPA of potential metabolites in serum (S) and urine (U); (B). PLS-DA scores plots for serum (S) and urine (U) samples from models, controls and treatment group in positive and negative ion mode.

FIGURE 4. The differentially expressed genes in the control and model groups of MA map (A), volcano map (B) and heat map (C). Analysis result of GO (D) between the control and model groups. KEGG enrichment (E) of the control and model groups.

FIGURE 5. (A,B). Power value filtering based on network; (C). Genetic module classification; (D). Heat map of correlation between module and biochemical index (”***”: p < 0.001; “**”: 0.001 ≤ p < 0.01; “*”: 0.01 ≤ p < 0.05; “.”: 0.05 ≤ p ≤ 0.1; P＞0.1, it will not be displayed); (E). Relative expression of candidate genes different groups of samples (# p < 0.05; ## p < 0.01; ### p < 0.001: models vs control; * p < 0.05; ** p < 0.01; *** p < 0.001: treatment groups vs. models). Real-time fluorescence quantitative PCR validation of candidate genes.

FIGURE 6. Quantitative comparison of target proteins. (ns: P ＞ 0.05, not significant; # p < 0.05; ## p < 0.01; ### p < 0.001: models vs. control; *:p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001)

FIGURE 7. Molecular mechanism of multi-bioactive compounds combination from Salvia miltiorrhiza to improve DKD. (The red arrows represent changes in db/db model mice).