Intervention Effects of Ganoderma Lucidum Spores on Epileptiform Discharge Hippocampal Neurons and Expression of Neurotrophin-4 and N-Cadherin

Abstract

Epilepsy can cause cerebral transient dysfunctions. Ganoderma lucidum spores (GLS), a traditional Chinese medicinal herb, has shown some antiepileptic effects in our previous studies. This was the first study of the effects of GLS on cultured primary hippocampal neurons, treated with Mg2+ free medium. This in vitro model of epileptiform discharge hippocampal neurons allowed us to investigate the anti-epileptic effects and mechanism of GLS activity. Primary hippocampal neurons from <1 day old rats were cultured and their morphologies observed under fluorescence microscope. Neurons were confirmed by immunofluorescent staining of neuron specific enolase (NSE). Sterile method for GLS generation was investigated and serial dilutions of GLS were used to test the maximum non-toxic concentration of GLS on hippocampal neurons. The optimized concentration of GLS of 0.122 mg/ml was identified and used for subsequent analysis. Using the in vitro model, hippocampal neurons were divided into 4 groups for subsequent treatment i) control, ii) model (incubated with Mg2+ free medium for 3 hours), iii) GLS group I (incubated with Mg2+ free medium containing GLS for 3 hours and replaced with normal medium and incubated for 6 hours) and iv) GLS group II (neurons incubated with Mg2+ free medium for 3 hours then replaced with a normal medium containing GLS for 6 hours). Neurotrophin-4 and N-Cadherin protein expression were detected using Western blot. The results showed that the number of normal hippocampal neurons increased and the morphologies of hippocampal neurons were well preserved after GLS treatment. Furthermore, the expression of neurotrophin-4 was significantly increased while the expression of N-Cadherin was decreased in the GLS treated group compared with the model group. This data indicates that GLS may protect hippocampal neurons by promoting neurotrophin-4 expression and inhibiting N-Cadherin expression.

Repeated Administration of Ketamine can Induce Hippocampal Neurodegeneration and Long-Term Cognitive Impairment via the ROS/HIF-1a Pathway in Developing Rats

“…To determine the percentage of neurons present, hippocampal neurons in culture were labeled by immunofluorescence staining, according to previous protocols [30]. Briefly, the neurons were cultured for 5 days, and the cultures were fixed with 4% paraformaldehyde for 15 min and were incubated with the neuron-specific primary antibody against neuron-specific enolase (NSE) (Abcam, Cambridge‚UK; 1:200) overnight and finally with an appropriate fluorophore-tagged secondary antibody.…”

Protective Effects of Sporoderm-Broken Spores of Ganderma lucidum on Growth Performance, Antioxidant Capacity and Immune Function of Broiler Chickens Exposed to Low Level of Aflatoxin B1

“…The spores of Ganoderma lucidum (SGL), ejected from the pileus of G. lucidum in the mature phase, are tiny and mist-like particles of about 6.5–8.0 × 9.6–12.6 μm enwrapped with an outer bilayer of sporoderm [12]. During the past two decades, because the spores could be collected on a large scale, SGL has attracted extensive interest as studies revealed that the spores possess many bioactive substances, including polysaccharides, unsaturated fatty acids, triterpenoids, nucleotides, ergostero and other bioactive ingredients [13,14]. In vitro, the polysaccharides of SGL had strong bioactivity in scavenging 1,1-Diphenyl-2-picrylhydrazyl and oxygen radicals [15].…”

A retrospective study of Ganoderma Lucidum Spore Powder for patients with epilepsy

“…In the search for an alternative epilepsy therapy, Ganoderma Lucidum Spore Powder (GLSP) is a potential intriguing candidate. [ 11 – 14 ] It has been reported that animal data have supported that the antiepileptic effect of GLSP in both in vivo and in vitro studies. [ 11 – 24 ] Presently, limited data of using GLSP in treating human epilepsy is available.…”