Neuroprotective Ganoderma compositions and methods of use

A technology of Ganoderma lucidum extract and Ganoderma lucidum, applied in] in one field, can solve the problems of weakening the inflammatory response of microglial cells, no report of Ganoderma lucidum, etc.

Inactive Publication Date: 2012-08-01
PURAPHARM INT HK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0012] However, it has not been reported that Ganoderma can attenuate the inflammatory response of microglia to exogenous or endogenous stimuli and / or protect dopaminergic neurons from invariance

Method used

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  • Neuroprotective Ganoderma compositions and methods of use
  • Neuroprotective Ganoderma compositions and methods of use
  • Neuroprotective Ganoderma compositions and methods of use

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0063] MES 23.5 Preparation of Cell Membrane Fractions

[0064] After MES 23.5 cells were exposed to MPP+10 μM for 24 hours, they were harvested in a medium containing 0.25M sucrose, 100mM PBS, 1mM MgCl 2 , 1mM EDTA and 2μM protease inhibitor PMSF buffer, and use glass-Teflon homogenizer (glass-teflon homogenizer) to homogenize it (Le, W.D.et al., 2001, J.Neurosci., 21: 8447-8455). The homogenate was then centrifuged at 8000 xg for 10 minutes at 4°C to remove the native nuclear fraction. The supernatant was centrifuged again at 100000 x g for 60 min at 4 °C. The pellet was homogenized and suspended in culture medium and used as the neuronal membrane fraction.

[0065] High affinity [3H] dopamine uptake assay

[0066] With 1ml Krebs-Ringer buffer (16mM NaH 2 PO 4 , 16mM Na 2 HPO 4 , 119mM NaCl, 4.7mM KCl, 1.8mM CaCl 2 , 1.2 mM MgSO 4 , 1.3 mM EDTA and 5.6 mM glucose; pH 7.4) washed the cells in each well. The cells were then incubated with 10 nM [3H]dopamine in Krebs...

Embodiment 1

[0078] Example 1-LPS and MPP+ treated dopaminergic membrane-induced microglia activation

[0079]To model microglial activation in neurodegeneration, LPS and MPP were used in microglial cultures or co-cultures of dopaminergic neurons (MES23.5 cell line) and microglia + Treated dopaminergic cell membranes serve as stimuli.

[0080] Microglia were visualized by staining for the CR3 complement receptor using the monoclonal antibody OX-42. The purity of the microglia culture was -95%. Non-activated microglia displayed branched shapes or bipolar or multipolar protrusions (Fig. 1a and b). Activated microglia exhibited an amoeboid-like morphology (Fig. 1c and d).

[0081] Among numerous neurotoxic factors, NO, TNF-α, IL-1β, and superoxide may be the main regulators of dopaminergic neurodegeneration induced by microglial activation. LPS-induced microglial activation was characterized by measuring TNF-α and IL-1β levels, two well-documented cytokines that reflect microglial activat...

Embodiment 2

[0085] Example 2 - Ganoderma lucidum prevents the production of pro-inflammatory factors and ROS derived from microglia

[0086] Microglia can produce cytokines as a result of activation (20-22). To elucidate the underlying mechanism of the neuroprotective activity of Ganoderma lucidum, the effects of Ganoderma lucidum on the levels of microglia-derived inflammatory cytokines and ROS were investigated. Microglial cultures were pretreated with different doses (50–400 μg / ml) of Ganoderma lucidum for 30 min before exposure to LPS or MPP + Processed CF.

[0087] Such as image 3 and 4 It was shown that low doses (50 μg / ml) of G. lucidum had minimal inhibitory effects, whereas pretreatment with higher doses (100–400 μg / ml) of G. lucidum potently reduced NO and CF induced by LPS or CF in a concentration-dependent manner. Increase of SOD.

[0088] At the same concentration, Ganoderma lucidum also significantly reduced exposure to LPS and MPP + Release of TNF-[alpha] and IL-1[be...

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Abstract

The subject invention provides methods for treatment of degenerative neurological disorders such as Parkinson's disease and Alzheimer's disease comprising administration of a Ganoderma lucidum extract. The subject invention also provides a method for inhibiting the activation of microglial cells by applying the Ganoderma lucidum extract to the cells.

Description

[0001] Cross references to related patent applications [0002] This application claims the benefit of US Provisional Application No. 61 / 173,802, filed April 29, 2009, which is incorporated herein by reference in its entirety. Background technique [0003] Parkinson's disease (PD) is a common neurodegenerative disorder that causes slowness of movement, stiffness, resting tremor, and loss of balance. As the disease progresses, many patients develop non-motor symptoms including anxiety, depression, constipation and dementia. [0004] Although there are drugs that can reduce the symptoms of PD, long-term use of these drugs is not effective in preventing the development of PD and brings debilitating side effects. There is therefore a great need to develop neuroprotective therapies aimed at slowing or even halting the progression of said neurodegeneration. [0005] Unfortunately, limited knowledge of the pathogenesis of degenerative neurological diseases such as PD hampers the d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/074A61P25/28A61P25/16A61P25/00
CPCA61K36/074A61P17/02A61P25/00A61P25/02A61P25/08A61P25/16A61P25/18A61P25/28A61P29/00A61P31/22A61P39/06A61P43/00A61P9/10
Inventor B·P·陈R·张
Owner PURAPHARM INT HK
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