Ginsenoside F1 attenuates pirarubicin-induced cardiotoxicity by modulating Nrf2 and AKT/Bcl-2 signaling pathways

Fig. 1. GF1's effects on THP-induced hemodynamic indices, electrocardiograms, and serum cardiac enzymes in rats. (A) Left ventricular systolic pressure (LVSP); (B) left ventricular end-diastolic pressure (LVEDP); (C and D) maximal left ventricular pressure change (±dp/dtmax); (E) cardiac QRS complex; (F) representative cardiac Q-T interval; (G) representative heart rate; (H) representative levels of lactate dehydrogenase (LDH); (I) representative levels of creatine kinase MB (CK-MB); (J) representative cardiac troponin T levels (c-TnT); and (K) representative brain natriuretic peptide levels (BNP). ∗P＜0.05 in comparison to the CON group; #P＜0.05 in comparison to the THP group; △P＜0.05 in comparison to the DZR group. Each data value represents mean ± SD of in each group (n =6).

Fig. 2. Effects of GF1 on rat cardiac oxidative stress and histological alterations produced by THP. (A) Superoxide dismutase (SOD) levels; (B) malondialdehyde (MDA) content; (C) glutathione (GSH) levels; (D) representative Nrf2 levels in the nucleus; in the nucleus Lamin B was employed as an internal control. Protein expression of Nrf2, keap1, HO-1, Gst, and GCLM in the cytoplasm; GAPDH was employed as an internal control in the cytoplasm; (E) protein ratios of Nrf2 and Lamin B in the nucleus; (F) protein ratios of Nrf2 and GAPDH in the cytoplasm; (G-J) protein ratios of cytosolic keap1, HO-1, Gst, and GCLM and GAPDH. ∗P＜0.05 in comparison to the CON group; #P＜0.05 in comparison to the THP group; △P＜0.05 in comparison to the DZR group. Each data value represents mean ± SD of in each group (n =3). (K) Staining using H&E (200× magnification; n =6); (L) HE staining is scored on a five-point scale from 0 to 4.

Fig. 3. THP-induced related-protein expression in rat cardiac tissue and the effects of GF1. (A) Immunohistochemistry pictures of cyt-c in cardiac tissue from rats; (B) Image-Pro Plus 6.0 software was used to assess the immunohistochemistry intensity of cyt-c expression; (C) Immunohistochemistry pictures of p-AKT in cardiac tissue from rats; (D) Image-Pro Plus 6.0 software was used to assess the immunohistochemical intensity of p-AKT expression; (E) Representative caspase 3 immunofluorescence images in rat cardiac tissue; (F) Image-Pro Plus 6.0 software was used to evaluate the fluorescence intensity of caspase 3; (G) Protein expression of the p-AKT, Bcl-2, Bax, and caspase families; GAPDH was used as an internal control; (H) p-AKT and AKT protein ratios; (I–K) representative caspase family and GAPDH ratios; (L) Bcl-2 and Bax ratios. ∗P＜0.05 in comparison to the CON group; #P＜0.05 in comparison to the THP group; △P＜0.05 in comparison to the DZR group. Each data value represents mean ± SD of in each group (n =3).

Fig. 4. H9c2 cells were protected from THP-induced apoptosis by GF1. (A) GF1 cytotoxicity on H9c2 cells. ∗p＜0.05 vs. 0 μM group (n =6); (B) THP effect on H9c2 cell survival rate. ∗p＜0.05 compared to the CON group (n =6); (C) representative of GF1 effects on H9c2 cell viability induced by THP (n =6); (D) representative p-AKT, Bcl-2, Bax, and caspase family protein expression; GAPDH was used as an internal control; (E) representative p-AKT and AKT protein ratios; (F–H) representative caspase family and GAPDH protein ratios; (I) representative Bcl-2 and Bax ratios. ∗P＜0.05 in comparison to CON group; #P＜0.05 in comparison to THP group; △P＜0.05 in comparison to GF1 group. Each data value represents mean ± SD of in each group (n =3).

Fig. 5. THP-induced oxidative stress in H9c2 cells and the effects of GF1. (A) Representative levels of superoxide dismutase (SOD); (B) representative levels of malondialdehyde (MDA); (C) representative levels of glutathione (GSH); (D) representative Nrf2 levels in nucleus; Lamin B was used as an internal control in nucleus; Nrf2, keap1, HO-1, Gst, and GCLM protein expression in cytoplasm; GAPDH was utilized as an internal control in cytoplasm; (E) representative nucleus Nrf2 and Lamin B protein ratio; (F) representative cytosolic Nrf2 and GAPDH protein ratios; and (G-J) representative cytosolic keap1, HO-1, Gst, and GCLM and GAPDH protein ratios. ∗P＜0.05 in comparison to CON group; #P＜0.05 in comparison to THP group; △P＜0.05 in comparison to GF1 group. Each data value represents mean ± SD of in each group (n =3).

Fig. 6. Scheme summarizes GF1 against THP-induced myocardial toxicity via activation Nrf2 and AKT/Bcl-2 signaling pathways to inhibit oxidative stress and apoptosis.