A defined multi-metabolite formulation derived from Salvia miltiorrhiza alleviates obesity-associated cardiac lipotoxicity via modulating mitochondrial dynamics

Figures

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  FIGURE 1. SABP increases H9c2 cell survival and improves mitochondrial function. (A) Chemical structures of the four metabolites in Salvia miltiorrhiza Bunge aqueous extract. (B) H9c2 cell viability following treatment with PA at different concentrations for 12, 24, 48 h tested by CCK-8 assay (n = 3). (C) H9c2 cell viability after treatment with 400 μM PA for 24 h and followed by different concentrations of SABP for 12, 24, and 48 h (n = 3). (D,E) Representative JC-1 fluorescence images and the Red/Green fluorescence ratio (n = 3 independent experiments; five to eight fields were analyzed per group per experiment). Scale bar: 50 μm. Data are expressed as mean ± SD. ## P < 0.01 vs. Control group. ** P < 0.01 vs. Model group.

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  FIGURE 2. SABP regulates mitochondrial dynamics and apoptotic protein expression in H9c2 cells. (A–G) Expression levels of Opa1, Mfn2, Drp1, Fis1 (A,C–F) and caspase-3, cleaved caspase-3 (B,G) in H9c2 cells determined by Western blot (n = 3). Data are expressed as mean ± SD. ## P < 0.01 vs. Control group. * P < 0.05, ** P < 0.01 vs. Model group.

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  FIGURE 3. SABP improves cardiac function and reduces fat accumulation in high-fat-diet induced mice. (A,C–F) Representative images of echocardiography and measurements of LVEF, LVFS, LVESD and LVEDD (n = 3). (B,G) Representative images of Micro-CT and fat volume-to-total volume ratio in mice (n = 3). (H) Ratio of heart weight to tibia length (n = 3). (I) Heart weight to body weight ratio (n = 3). Data are expressed as mean ± SD. ## P < 0.01 vs. Control group. * P < 0.05, ** P < 0.01 vs. Model group.

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  FIGURE 4. SABP ameliorates dyslipidemia and inflammatory response and alleviates myocardial injury in high-fat diet-induced mice. (A–D) Serum lipid and TG, TC, LDL-C, HDL-C levels in mice. (E–H) Levels of IL-1β, IL-6, TNF-α and IL-10 in serum. (I,J) Levels of LDH and CK-MB in serum. Data are expressed as mean ± SD. # P < 0.05, ## P < 0.01 vs. Control group. * P < 0.05, ** P < 0.01 vs. Model group.

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  FIGURE 5. SABP alleviates myocardial tissue damage and mitochondrial ultrastructural abnormalities in high-fat-induced mice. (A,B) Representative H&E (A) and Masson’s trichrome staining (B) of the left ventricular apex. Scale bar: 50 μm. (C) Representative TEM images of damaged mitochondria. Scale bars: top row, 2 μm; bottom row, 1 μm. (D) Quantitative analysis of the number of mitochondria per μm2 (n = 3). (E) Quantitative analysis of the mean mitochondrial size (n = 3). Data are expressed as mean ± SD. ## P < 0.01 vs. Control group. * P < 0.05, ** P < 0.01 vs. Model group.

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  FIGURE 6. SABP regulates mitochondria-related protein expression and inhibits myocardial apoptosis to alleviate high-fat-induced myocardial injury in mice. (A,C–F) Protein expression of Opa1, Mfn2, Drp1, Fis1 in cardiac tissue (n = 3). (B,G) Apoptosis related protein expression detected by Western blot and the ratio of cleaved-caspase 3/caspase-3 (n = 3). Data are expressed as mean ± SD. ## P < 0.01 vs. Control group. * P < 0.05, ** P < 0.01 vs. Model group.