Kaempferol Protects Against Apoptosis in PC12 Cells Exposed to Hydrogen Peroxide by Activating Akt1.

Objective: Spinal cord injury (SCI) is a debilitating disease that cannot be cured at present. Kaempferol (KPL) has proven neuroprotective, antioxidant, and antibacterial properties. However, the neuroprotective effect of KPL on SCI and its potential mechanism are still unclear. Methods: Network pharmacology and molecular docking were used to determine the potential mechanisms of KPL on SCI. In vitro studies were used to validate the results. Hydrogen peroxide (H₂O₂)-exposed PC12 cells were pretreated with or without KPL. The safe concentrations of KPL, H₂O₂, and MK2206 in PC12 cells were determined via cell counting kit-8 assays. Then, terminal deoxynucleotidyl transferase dUTP nick end labeling staining and Annexin V-FITC/PI double staining were used to detect apoptotic cells. In addition, Western blotting was utilized to measure apoptosis-related gene and protein expression levels. Results: Network pharmacology and molecular docking data demonstrated an interaction between KPL and Akt1 and identified apoptosis as a potential target of the KPL/Akt1 complex. The data verified that KPL could inhibit mitochondrial apoptosis in vivo by inducing Akt1 phosphorylation. Subsequently, MK2206, a pharmacological Akt1 inhibitor, was used to assess the role of Akt1 in this process, and we found that MK2206 attenuated the antiapoptotic effect of KPL on PC12 cells. Conclusion: This study demonstrates that KPL could inhibit mitochondrial apoptosis by inducing the phosphorylation of Akt1, suggesting that KPL is a very promising candidate for SCI treatment. [ABSTRACT FROM AUTHOR]