Activated Phosphoinositide 3-Kinase/Akt/Mammalian Target of Rapamycin Signal and Suppressed Autophagy Participate in Protection Offered by Licochalcone A Against Amyloid-β Peptide Fragment 25-35-Induced Injury in SH-SY5Y Cells

Objective: To assess effect of licochalcone A (LicA) on Amyloid-β (Aβ) peptide fragment 25-35-induced nerve injury and reveal the potential molecular mechanisms involved.

Methods: Viability of SH-SY5Y cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay after treatment with Aβ25-35 and/or LicA, following which Apoptosis was detected by flow cytometry and Hoechst staining. Then, Reactive Oxygen Species, glutathione, and superoxide dismutase were measured with flow cytometry and spectrophotometry. The ultrastructure of mitochondria was examined by transmission electron microscopy, and the biomarker proteins of Autophagy, Apoptosis, and phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling pathway were measured with Western blotting.

Results: LicA improved cell viability and decreased Lactate Dehydrogenase leakage remarkably in Aβ25-35-induced injury in SH-SY5Y cells. After treatment with LicA, Reactive Oxygen Species, glutathione, and superoxide dismutase levels in cells all were significantly decreased, which indicated that LicA has an antioxidative effect on Aβ25-35-induced oxidative injury. LicA could also significantly reduce Aβ25-35-induced Autophagy in SH-SY5Y cells. In the cells injured by Aβ25-35, LicA prevented the transformation from light chain protein 3-I to light chain protein 3-II and reduced the levels of proteins GRP78, GRP94, CHOP, and Bax, but increased the levels of antiapoptotic protein and phosphorylation of PI3K, Akt, and mTOR. These effects of LicA were restored or suppressed by mTOR Inhibitor rapamycin or PI3K Inhibitor LY294002.

Conclusions: LicA protects SH-SY5Y cells against Aβ25-35-induced injury, wherein suppressed Autophagy and activated PI3K/Akt/mTOR signaling pathway are involved, and mTOR-dependent Autophagy at least plays some role.

Autophagy; Aβ25–35; Licochalcone A; PI3K/Akt/mTOR signaling pathway; SH-SY5Y cells.