Berberine Protects C17.2 Neural Stem Cells From Oxidative Damage Followed by Inducing Neuronal Differentiation

Jia-Wen Shou; Chun-Kai Cheung; Jian Gao; Wei-Wei Shi; Pang-Chui Shaw

doi:10.3389/fncel.2019.00395

Berberine Protects C17.2 Neural Stem Cells From Oxidative Damage Followed by Inducing Neuronal Differentiation

Front Cell Neurosci. 2019 Sep 3:13:395. doi: 10.3389/fncel.2019.00395. eCollection 2019.

Authors

Jia-Wen Shou^{1

2}, Chun-Kai Cheung², Jian Gao³, Wei-Wei Shi³, Pang-Chui Shaw^{1

2

4}

Affiliations

¹ School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong.
² Li Dak Sum Yip Yio Chin R&D Centre for Chinese Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong.
³ Shenzhen Health Development Research Center, Shenzhen, China.
⁴ State Key Laboratory of Research on Bioactivities and Clinical Applications of Medicinal Plants and Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong.

Abstract

Neurodegeneration is the loss of structure and/or function of neurons. Oxidative stress has been suggested as one of the common etiology in most of the neurodegenerative diseases. Previous studies have demonstrated the beneficial effects of berberine in various neurodegenerative and neuropsychiatric disorders. In this study, we hypothesized that berberine could protect C17.2 neural stem cells (NSCs) from 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative damage then promote neuronal differentiation. AAPH was used to induce oxidative damage. After the damage, berberine protected C17.2 cells were kept cultured for another week in differentiation medium with/without berberine. Changes in cell morphology were detected by microscopy and cell viability was determined by MTT assay. Real-time PCR and western blot analysis were performed to confirm the associated pathways. Berberine was able to protect C17.2 NSCs from the oxidative damage. It lowered the cellular reactive oxygen species (ROS) level in C17.2 cells via Nuclear Factor Erythroid 2-Related Factor 1/2 (NRF1/2) - NAD(P)H Quinone Dehydrogenase 1 (NQO-1) - Heme Oxygenase 1 (HO-1) pathway. It also down-regulated the apoptotic factors-Caspase 3 and Bcl2 Associated X (Bax) and upregulated the anti-apoptotic factor-Bcl2 to reduce cell apoptosis. Besides, berberine increased C17.2 cell viability via up-regulating Extracellular-signal-Related Kinase (ERK) and phosphor-Extracellular-signal-Related Kinase (pERK) expression. Then, berberine promoted C17.2 cell to differentiate into neurons and the differentiation mechanism involved the activation of WNT/β-catenin pathway as well as the upregulation of expression levels of pro-neural factors Achaete-Scute Complex-Like 1 (ASCL1), Neurogenin 1 (NeuroG1), Neuronal Differentiation 2 (NeuroD2) and Doublecortin (DCX). In conclusion, berberine protected C17.2 NSCs from oxidative damage then induced them to differentiate into neurons.

Keywords: berberine; differentiation; neural stem cell; neuron; oxidative damage.