Neuroprotection of isookanin against MPTP-induced cell death of SH-SY5Y cells via BCL2/BAX and PI3K/AKT pathways.

BACKGROUND AND PURPOSE

Isookanin, an important antioxidant component in Coreopsis tinctoria Nutt., has shown remarkable hypolipidemic, hypoglycemic, and hypotensive effects. However, the neuroprotective effect of isookanin has not been reported yet. Here, the neuroprotective effects and relevant molecular mechanisms of isookanin are explored for the first time.

METHODS

The SH-SY5Y cells were exposed to neurotoxic HO, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and Aβ, respectively. Cell viability and apoptosis were evaluated by MTT, lactate dehydrogenase (LDH), and TUNEL assays. Intercellular ROS and mitochondrial membrane potential were assessed by DCFH-DA and JC-1 assay. Western blot and qRT-PCR were used to explore the perturbed signaling at the gene and protein levels. Molecular docking analysis and in vitro assay were further applied to confirm potential target.

RESULTS

Among the three in vitro models, isookanin showed the best neuroprotection against MPTP-induced damage. Isookanin attenuated the levels of LDH, intracellular ROS, and mitochondrial membrane potential. Isookanin upregulated phosphorylation of AKT and PI3K, and increased BCL2/BAX ratio. Isookanin possessed a powerful affinity toward AKT. Besides, the protective effects of isookanin disappeared when cells were co-treated with an AKT inhibitor (AZD5363).

CONCLUSION

Isookanin regulated BCL2/BAX and PI3K/AKT pathways to reduce mitochondrial damage and cellular apoptosis. Isookanin may be a new protector for neurodegenerative diseases.