Sodium Glucose Cotransporter 2 (SGLT2) Inhibitor Ameliorate Metabolic Disorder and Obesity Induced Cardiomyocyte Injury and Mitochondrial Remodeling

Figure 1. Effects of the pericardial fat secretome of different groups of metabolic-syndrome-associated mice on cardiomyocyte H9c2. Secretome of pericardial fat on H9c2 cell viability inhibition at (A) 48 h and (B) 72 h. (C) The effect of secretome on mitochondrial membrane potential alteration of H9c2. 50 ug/mL CCCP was used as positive control. (D) ATP production capacity and (E) NAD+/NADH ratio were determined as the evaluation of H9c2 mitochondrial function after treatment with different groups’ pericardial fat secretome. (F) Rhod-2 AM staining for intra-mitochondria Ca2+ distribution (Magnification: 400×). In vitro study, n = 3 for each group, * p < 0.05 compared with control group, # p < 0.05 compared with MDO group, Δ p < 0.05 compared with EMPA group.

Figure 2. ROS production and antioxidant expression in response to oxidative stress in H9c2 cells. Effects of epicardial fat secretome on the production and accumulation of (A) cellular ROS and (B) intra-mitochondria ROS of H9c2 cells. (Magnification: 400×) (C) The OS-associated antioxidant SOD1 and SOD2 genes in H9c2 cells respond to the secretome. (D) Protein expression changes and (E) quantitative results of antioxidants SOD1 and SOD2 in H9c2 cells. In vitro study, n = 3 for each group, # p < 0.05 compared with MDO group.

Figure 3. MDO- and EMPA-associated epicardial fat secretome in genes regulation correlated with mitochondrial function. (A) The expression alteration of regulatory genes related to mitochondrial fission and fusion of H9c2 treated by different epicardial fat secretomes. Western blot analysis of (B) protein expression and (C) the quantitative results of regulators involved in secretome-manipulated H9c2 mitochondrial function. (D) The impact of fat secretomes on the gene expression of Ca2+ channel regulators in mitochondria. # p < 0.05 compared with MDO group (E) KEGG pathway analysis of RNA-sequencing predictions’ potential gene network affected by the epicardial fat secretome. (In vitro study, n = 3 for each group).

Figure 4. RNA-sequencing analysis of the impact of epicardial fat secretomes on H9c2 cellular process. (A) Volcano pots of differentially expressed genes (DEGs). (B) Canonical Pathway View and (C) Disease & Functions comparative analysis of DEGs’ participation. The analysis of DEGs in (D) cellular component (CC) and (E) biological process (BP) involved in cellular process were revealed. The gene regulatory network of (F) CC and (G) BP were shown by emap plot. (In vitro study, n = 3 for each group).

Figure 5. HFD-mouse model for verification of protein expression associated with mitochondrial regulation after treatment (n = 5 for each group). IHC histological of HFD mouse model for the analysis of (A) SOD1, (B) SOD2, (C) ifit-1, (D) mfn2, (E) Sirt1, and (F) Sirt3 expression regulated in individual mouse group. Left panel for left ventricle and right panel for right ventricle of cardiac tissue. Magnification: 200×.