Withdrawn: Safranal induces autophagy by AMPK activation and protects neurons against amyloid beta in Alzheimer’s disease

Xiaocheng Huang,Ying Hua,Manlian Zhu,Xiumei Yan,Ruilai Jiang

doi:10.4314/tjpr.v18i3.2

Abstract

This article has been withdrawn by the Editor Purpose: To investigate autophagic induction by safranal and neuroprotection against amyloid beta in Alzheimer’s disease.  Methods: Primary neurons and SH-SY5Y cells were used in this study. Assessment of cell proliferation and neuroprotection by safranal against amyloid beta was done by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay. AMPK activation and mTOR inhibition were determined by western blot. Changes in intracellular calcium level, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were assessed by flow cytometry.  Results: Safranal protected neurons against amyloid beta toxicity. Furthermore, safranal activated AMPK pathway by activation of calcium/calmodulin-dependent protein kinase (CaMKKβ) to induce autophagy in both cell lines. The toxicity induced by amyloid beta in primary neurons and SH-SY5Y cells were attenuated by safranal. Moreover, amyloid beta-induced calcium levels were significantly decreased by safranal while ROS and MMP loss produced by amyloid beta was attenuated by safranal.  Conclusion: These findings suggest that safranal protects neurons against amyloid beta by inducing autophagy via AMPK pathway. Therefore, safranal is a probable therapeutic target for Alzheimer’s disease.

Highlights

Alzheimer’s disease (AD) is a neurodegenerative disease which happens in old age [1]
Neuroprotection assay was performed by MTT assay in which differentiated SH-SY5Y cells and primary neurons were grown in 96 well plates and were pre-treated with safranal for 24 h at a concentration of 5, 10, 20, 40, 60, 80 and 100 μM followed by amyloid beta (20 μM)
Safranal protected SH-SY5Y cells and primary neuronal cells at concentrations of 5, 10, 20, 40 and 60 μM against amyloid beta induced toxicity as the viability of cells treated with safranal only increased for 48 h (Figure 1 A)

Summary

INTRODUCTION

Alzheimer’s disease (AD) is a neurodegenerative disease which happens in old age [1]. For drug treatments in case of neuroprotection assay, cells at a confluence of 70 % was pre-treated with safranal followed by amyloid beta treatment at indicated time periods and concentrations. Neuroprotection assay was performed by MTT assay in which differentiated SH-SY5Y cells and primary neurons were grown in 96 well plates and were pre-treated with safranal for 24 h at a concentration of 5, 10, 20, 40, 60, 80 and 100 μM followed by amyloid beta (20 μM). Cell viability assay was performed by MTT assay, in which differentiated SH-SY5Y cells and primary neurons were grown in 96 well plates These cells were treated with safranal at a concentration of 1, 5, 10, 20, 40, 60, 80 and 100 μM for 48 h and 72 h.

RESULTS

DISCUSSION

Conflicts of interest