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Aβ peptide secretion is reduced by Radix Polygalaeinduced autophagy via activation of the AMPK/mTOR pathway

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Radix Polygalae is a traditional Chinese medicine that has been used as a sedative and to improve memory for a number of years. The impact of Radix Polygalae in patients with Alzheimer's disease has been investigated. However the mechanisms underlying its effects remain unclear. In the current study, the toxicity of various doses (100, 40, 20, 10, 5 and 0 µg/ml) of Radix Polygalae was measured in the human neuroblastoma cell line (SHSY5Y) using an MTT assay. Changes in amyloid β (Aβ) levels in the supernatant of Chinese hamster ovary (CHO) cells overexpressing βamyloid proprotein (APP) and BACE1 (CHOAPP/BACE1), were detected using an ELISA assay. In order to confirm that the Aβ reduction was associated with autophagy, the autophagy marker protein, light chain 3 (LC3), was measured by western blot analysis and autophagosomes were assessed using MDC staining. In addition, the mechanism underlying the autophagy induced by Radix Polygalae was analyzed using western blotting to measure the protein expression of mammalian target of rapamycin (mTOR), p70s6k, Raptor, protein kinase B and adenosine monophosphateactivated protein kinase (AMPK), in addition to the phosphorylated forms of these proteins. The results demonstrated no significant toxicity of Radix Polygalae in SHSY5Y cells, at a dose of 100 µg/ml. The secretion of Aβ was markedly reduced following treatment with Radix Polygalae, and this reduction occurred in a dosedependent manner. The autophagy levels were shown to be enhanced in the drug treatment group, using fluorescence microscopy. In addition, levels of LC3/LC3, the marker protein of autophagy, were also increased. The results of the current study suggest that Radix Polygalae may aid in the elimination of the Aβ peptide, via the induction of autophagy, by the AMPK/mTOR signaling pathway. These results may provide a basis for further kin vivo investigation.
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Document Type: Research Article

Affiliations: 1: Department of Histology and Embryology, Anhui Medical University, Hefei, Anhui 230032, P.R. China 2: Department of Laboratorial Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200071, P.R. China 3: Shanghai Genhouse Technology Co., Ltd., Shanghai 201203, P.R. China

Publication date: August 1, 2015

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  • Molecular Medicine Reports is a monthly, peer-reviewed journal available in print and online, that includes studies devoted to molecular medicine, underscoring aspects including pharmacology, pathology, genetics, neurosciences, infectious diseases, molecular cardiology and molecular surgery. In vitro and in vivo studies of experimental model systems pertaining to the mechanisms of a variety of diseases offer researchers the necessary tools and knowledge with which to aid the diagnosis and treatment of human diseases.
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