Huperzine A Alleviates Oxidative Glutamate Toxicity in Hippocampal HT22 Cells via Activating BDNF/TrkB-Dependent PI3K/Akt/mTOR Signaling Pathway

Abstract

Oxidative glutamate toxicity is involved in diverse neurological disorders including epilepsy and ischemic stroke. Our present work aimed to assess protective effects of huperzine A (HupA) against oxidative glutamate toxicity in a mouse-derived hippocampal HT22 cells and explore its potential mechanisms. Cell survival and cell injury were analyzed by MTT method and LDH release assay, respectively. The production of ROS was measured by detection kits. Protein expressions of BDNF, phosphor-TrkB (p-TrkB), TrkB, phosphor-Akt (p-Akt), Akt, phosphor-mTOR (p-mTOR), mTOR, phosphor-p70s6 (p-p70s6) kinase, p70s6 kinase, Bcl-2, Bax, and β-actin were assayed via Western blot analysis. Enzyme-linked immunosorbent assay was employed to measure the contents of nerve growth factor, brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4 (NT-4). Our findings illustrated 10 μM HupA for 24 h significantly protected HT22 from cellular damage and suppressed the generation of ROS. Additionally, after treating with LY294002 or wortmannin [the selective inhibitors of phosphatidylinositol 3 kinase (PI3K)], HupA dramatically prevented the down-regulations of p-Akt, p-mTOR, and p-p70s6 kinase in HT22 cells under oxidative toxicity. Furthermore, it was observed that the protein levels of BDNF and p-TrkB were evidently enhanced after co-treatment with HupA and glutamate in HT22 cells. The elevations of p-Akt and p-mTOR were abrogated under toxic conditions after blockade of TrkB by TrkB IgG. Cellular apoptosis was significantly suppressed (decreased caspase-3 activity and enhanced Bcl-2 protein level) after HupA treatment. It was concluded that HupA attenuated oxidative glutamate toxicity in murine hippocampal HT22 cells via activating BDNF/TrkB-dependent PI3K/Akt/mTOR signaling pathway.

MicroRNA-382 inhibits cell proliferation and invasion of retinoblastoma by targeting BDNF-mediated PI3K/AKT signalling pathway

“…miR-382, microRNA-382; miR-NC, miRNA mimic negative control; RT-qPCR, reverse transcription quantitative polymerase chain reaction; CCK-8, Cell Counting Kit-8.tumour-suppressing roles in RB, at least in part by regulating BDNF.miR-382 inhibits PI3K/AKT signalling in RB. BDNF is involved in PI3K/AKT signalling pathway regulation(35,36).…”

Schisandrae Chinensis Fructus inhibits behavioral deficits induced by sleep deprivation and chronic unpredictable mild stress via increased signaling of brain‐derived neurotrophic factor

“…Our data showed that SD (12 hr) and SCF could upregulate TrkB/CREB/ERK signaling, which suggest that the increased BDNF mitigates depressive symptoms mainly by binding to TrkB, leading to autophosphorylation of TrkB tyrosine residues and activation of downstream signaling molecules, including the ERK known to phosphorylate CREB. In addition, BDNF/TrkB downstream signaling includes activation of Ras/ERK and PI3K/AKT pathways (Luo et al, ; Mao, Zhou, Li, & Liu, . Basic and clinical studies demonstrated that BDNF related PI3K/AKT signaling pathway was involved in the stress‐induced depression and antidepressant‐induced improvement (Wang et al, ; Zeng et al, ).…”

Acetylcholinesterase-independent protective effects of huperzine A against iron overload-induced oxidative damage and aberrant iron metabolism signaling in rat cortical neurons

“…Although the protective effects of HupA, a highly selective and potent AChE inhibitor, against neuronal damage have been well proven in multiple in vitro neuronal models [26,30,38,39] , whether and how this active natural product can directly ameliorate iron overload-induced neuronal damage has not yet been clarified. The most notable aspects of the present study include: 1) the significant effect of HupA on alleviating multifaceted iron overload-induced malignant changes in primary cortical neurons, and 2) the finding that the classical AChEinhibiting effect of HupA may not contribute to its protection against iron-overload-induced neuronal damage.…”

“…Interestingly, increasing numbers of studies have demonstrated that HupA can ameliorate oxidative stress and mitochondrial dysfunction [20][21][22] . First, HupA significantly suppresses ROS overproduction and lipid peroxidation in Aβ- [23,24] , H 2 O 2 - [25] and glutamate [26] -stimulated cellular models. Second, HupA also markedly ameliorates oxidative stress in chronic cerebrally hypoperfused and aged rats [27,28] and attenuates mitochondrial dysfunction in APP/PS1 transgenic mice [29] and rats after intracerebral hemorrhage (ICH) [30] .…”