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Carnosic Acid Prevents Beta-Amyloid-Induced Injury in Human Neuroblastoma SH-SY5Y Cells via the Induction of Autophagy

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Abstract

Beta-amyloid (Aβ), the hallmark protein in Alzheimer’s disease (AD), induces neurotoxicity that involves oxidative stress and mitochondrial dysfunction, leading to cell death. Carnosic acid (CA), a polyphenolic diterpene isolated from the herb rosemary (Rosemarinus officinalis), was investigated in our study to assess its neuroprotective effect and underlying mechanism against Aβ-induced injury in human neuroblastoma SH-SY5Y cells. We found that CA pretreatment alleviated the Aβ25–35-induced loss of cell viability, inhibited both Aβ1–42 accumulation and tau hyperphosphorylation, reduced reactive oxygen species generation, and maintained the mitochondrial membrane potential. Moreover, CA increased the microtubule-associated protein light chain 3 (LC3)-II/I ratio and decreased SQSTM1(p62), indicating that CA could induce autophagy. Autophagy inhibitor 3-methyladenine (3-MA) attenuated the neuroprotective effect of CA, suggesting that autophagy was involved in the neuroprotection of CA. It was also observed that CA activated AMP-activated protein kinase (AMPK) but inhibited mammalian target of rapamycin (mTOR). Furthermore, blocking AMPK with si-AMPKα successfully inhibited the upregulation of LC3-II/I, prevented the downregulation of phosphorylation of mTOR and SQSTM1(p62), indicating that CA induced autophagy in SH-SY5Y cells via the activation of AMPK. These results suggested that CA might be a potential agent for preventing AD.

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Abbreviations

Aβ:

Beta-amyloid

AD:

Alzheimer’s disease

CA:

Carnosic acid

ROS:

Reactive oxygen species

MMP:

Mitochondrial membrane potential

LC3:

Microtubule-associated protein light chain 3

3-MA:

3-Methyladenine

AMPK:

AMP-activated protein kinase

mTOR:

Mammalian target of rapamycin

NFT:

Neurofibrillary tangles

CMA:

Chaperone mediated autophagy

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide

DMSO:

Dimethyl sulfoxide

RT:

Room temperature

PBS:

Phosphate buffered saline

PI:

Propidium iodide

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Acknowledgments

This work was supported by the Natural Science Foundation of China from Yong-Ning Deng (No.81301097).

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Authors and Affiliations

  1. Department of Neurology, The First Affiliated Hospital of Xi’an Jiaotong University, 277 West Yanta Rd, Xi’an, 710061, China

    Jie Liu & Qiu-Min Qu

  2. Department of Anesthesia and Perioperative Care, Center for Cerebrovascular Research, University of California, San Francisco, San Francisco, CA, USA

    Hua Su

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  1. Jie Liu
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Correspondence to Qiu-Min Qu.

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Liu, J., Su, H. & Qu, QM. Carnosic Acid Prevents Beta-Amyloid-Induced Injury in Human Neuroblastoma SH-SY5Y Cells via the Induction of Autophagy. Neurochem Res 41, 2311–2323 (2016). https://doi.org/10.1007/s11064-016-1945-6

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