Protective effects of traditional Chinese medicines Aconiti Lateralis Radix Praeparata and Angelicae Sinensis on H/R cardiomyocytes via the PI3K/Akt/GSK-3β pathway and autophagy pathway.

[BACKGROUND] Aconiti Lateralis Radis Praeparata (FZ, Fuzi) and Angelicae Sinensis Radix (DG, Danggui) are widely used in traditional Chinese medicine for the treatment of coronary heart disease. However, the protective effects of their combination on cardiomyocytes remain unexplored. This study aimed to investigate whether the combined application of these 2 herbs confers protective benefits on H9c2 cardiomyocytes subjected to hypoxia/reoxygenation (H/R) injury.

[METHODS] A H/R model (6h hypoxia followed by 6h reoxygenation) was established using H9c2 cardiomyocytes. The cells were randomly assigned to the following groups: control, model, FZ, DG, FZDG 3 mg/mL, FZDG 9 mg/mL, FZDG 27 mg/mL, FZDG 27 mg/mL + LY, and LY + model. Cell viability was assessed using the cell counting kit-8 assay, and apoptosis was measured by Annexin V-FITC/PI staining. The levels of lactate dehydrogenase, malondialdehyde, and superoxide dismutase were determined using colorimetric methods. The distribution and expression of apoptosis-related proteins were evaluated via immunohistochemistry. Protein and mRNA expression levels of key components in the phosphatidylinositol 3-kinase/protein kinase B/glycogen synthase kinase-3β (PI3K/Akt/GSK-3β) and autophagy pathways were analyzed by Western blot and reverse transcription-polymerase chain reaction, respectively.

[RESULTS] This study demonstrated that the FZDG 27 mg/mL treatment significantly enhanced cell viability and reduced the overall rate of apoptosis. This group also ameliorated oxidative stress injury in H/R cardiomyocytes, as evidenced by decreased levels of lactate dehydrogenase and malondialdehyde, along with an increase in superoxide dismutase activity. Moreover, the expression and distribution of key proteins were modulated, with elevated phosphorylation levels of Akt and GSK-3β, as well as altered expression of B-cell lymphoma-2 and B-cell lymphoma-2-associated X protein. Western blot and reverse transcription-polymerase chain reaction demonstrated that the FZDG 27 mg/mL group most potently upregulated the PI3K/Akt/GSK-3β and autophagy pathways among all groups, an effect blocked by LY294002.

[CONCLUSION] This study demonstrated that the combination of Aconiti Lateralis Radix Praeparata and Angelicae Sinensis significantly protected H/R cardiomyocytes in vitro. This protective effect was achieved through enhancing cell viability, reducing oxidative stress-induced injury, and modulating the expression and distribution of key proteins and genes. The underlying mechanisms involved the activation of the PI3K/Akt/GSK-3β signaling pathway and the autophagy pathway, as confirmed by the inhibitory effect of LY294002.