Huperzine a provides neuroprotection against several cell death inducers usingin vitro model systems of motor neuron cell death

Abstract

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease resulting from the progressive loss of motor neurons in the spinal cord and brain. To date, clinically effective neuroprotective agents have not been available. The current study demonstrates for the first time that huperzine A, a potential neuroprotective agent, has the ability to protect a motor neuron-like cell line and motor neurons in spinal cord organotypic cultures from toxin-induced cell death. The neuroblastoma-spinal motor neuron fusion cell line, NSC34 and rat spinal cord organotypic cultures (OTC) were exposed to cell death inducers for 24 h or 14 d, respectively, with and without pre-treatment with huperzine A. The inducers used here include: staurosporine, thapsigargin, hydrogen peroxide (H2O2), carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and L-(-)-threo-3-hydroxyaspartic acid (THA). These agents were selected as they induce apoptosis/necrosis via mechanisms implicated in patients with generalized motor neuron disease. Cell death was determined in NSC34 cells by metabolic activity, caspase activity/expression and by nuclear morphology and in the OTCs, using immunohistochemistry and Western blot analysis. Nuclear staining of NSC34 cells revealed cell death induced by staurosporine, thapsigargin, H2O2 and CCCP. This induction was significantly reduced with 2 h pre-treatment with 10 microM huperzine A (maximum, 35% rescue; p 0.05) following exposure to staurosporine, thapsigargin and H2O2 but not with CCCP. These data were supported by the metabolic assays and caspase activity. In addition, pre-treatment with huperzine A dramatically improved motor neuron survival, based on choline acetyltransferase (ChAT) expression analysis in OTCs following exposure to THA, and compared to THA-treated control cultures. These studies are currently being extended to include other inducers and with additional compounds as potential drug therapies that could be used in combination for the treatment of patients with ALS.

Examining the Interactome of Huperzine A by Magnetic Biopanning

“…In addition to be a potent inhibitor of AChE [3], studies had suggested Hup A may also exert its neuroprotective effects through many pathways, such as antioxidation [35]–[37], antiapoptosis [4]–[6], attenuating the metabolism of the amyloid precursor protein(APP) [8], [9], protecting Ischemia Injury [10]. Based on our data and also some earlier pharmacological studies [33], [34], we think there should be some interaction between Hup A and mitochondria proteins.…”

“…It is a potent inhibitor of acetylcholinesterase(AChE) [3]. However, many recent studies have also suggested that it may have other mechanisms including cell protection against apoptosis through reversing the down-regulation of the expression of Bcl-2 and up-regulation of the expressions of Bax and P53 [4], [5], [6], mitochondria protection against dysfunction by preserving major mitochondria enzymes activity and reducing reactive oxygen species (ROS) production [7], interfering with amyloid precursor protein (APP) cleavage [8], [9], etc [10]. However, the detailed molecular mechanisms of most of these pharmacology effects were still not clear.…”

A Review of Experimental Research on Herbal Compounds in Amyotrophic Lateral Sclerosis

“…Hup A displays a potent inhibitory activity through blocking the combination of the NMDA receptor with glutamate to prevent glutamate toxicity. Several inducers were used including H 2 O 2 , carbonyl cyanide m ‐chlorophenyl hydrazone (a mitochondrial uncoupler leading to de‐energization of mitochondria) and l ‐(‐)‐threo‐3‐hydroxyaspartic acid (a compound that triggers excitotoxicity) to kill neurons, and it was illustrated that Hup A was neuroprotective for motor neurons and may be useful as a complementary therapy for patients with ALS (Hemendinger et al ., ). Researchers will further examine the use of Hup A, alone or in combination with other compounds, in models to determine its efficacy as a potential treatment for ALS.…”

Enzyme Inhibitors Involved in the Treatment of Alzheimer’s Disease

“…It also possesses protective action against glutamate induced excitotoxicity in neurons [121]. It has shown protective action in neuroblastoma-spinal motor neuron fusion cell line and rat spinal cord organotypic cultures against various cell death inducers such as staurosporine, thapsigargin, hydrogen peroxide, carbonyl cyanide m-chlorophenyl hydrazone and L-(-)-threo-3-hydroxyaspartic acid (THA) [122].…”