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Ginsenoside Rg1 facilitates neural differentiation of mouse embryonic stem cells via GR-dependent signaling pathway
Authors:Jiaying Wu  Zongfu Pan  Meiyuan Cheng  Yuanyuan Shen  Hao Yu  Qiongyue Wang  Yijia Lou
Affiliation:1. Division of Cardio–Cerebral Vascular and Hepatic Pharmacology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China;2. Department of Pharmacology, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China;3. NeuroSurgery Department, 2nd Affiliated Hospital at School of Medicine, Zhejiang University, Hangzhou 310009, China
Abstract:
Ginsenoside Rg1, a steroidal saponin of high abundance in ginseng, possesses the neuroprotective effects. In this study, we tried to explore the effect of Rg1 on promoting differentiation of mouse embryonic stem (ES) cells towards the neuronal lineage and its potential role involved in glucocorticoid receptor (GR) activation. Rg1 treatment induced a remarkable increase in the population of neuron-like cells in a time-dependent manner. More than 80% of Rg1-treated embryoid bodies (EBs) differentiated into neuron-like cells on d 8 + 10. Furthermore, the gradually increased protein expression of neurofilament (NEFM) and β-tubulin III (a neuronal specific protein) was determined. GR expression gradually increased during the differentiation course. RU486, an antagonist of GR, could efficiently block the neurogenesis-promoting activity of Rg1. On the other side, Rg1 stimulated the phosphorylation of ERK1/2 and Akt at different time points through GR activation-dependent mechanisms. Treatment of both U0126 (an inhibitor of MEK) and LY294002 (an inhibitor of PI3 K), hampered the neuronal differentiation induced by Rg1. Meantime, U0126 further decreased Rg1-induced p-Akt expression. In conclusion, Rg1 possesses the effects on inducing differentiation of mouse ES cells into neurons in vitro via the GR-MEK-ERK1/2-PI3 K-Akt signaling pathway.
Keywords:DMSO, dimethylsulfoxide   Dex, dexamethasone   EB, embryoid body   ERK, extracellular signal-related protein kinases   ES, embryonic stem   GAPDH, glyceraldehyde-3-phosphate dehydrogenase   JNK, c-Jun N-termianl kinases   LIF, leukemia inhibitory factor   MAPK, mitogen-activated protein kinase   MEF, mouse embryonic fibroblast   NEAA, non-essential amino acid   NEFM, neurofilament   PI3 K, phosphatidylinositol 3-kinase   RA, retinoic acid   Rg1, ginsenoside Rg1   RU486, mifepristone
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