CN104958314B - Sparassis crispa polyoses extract is preparing the purposes for the treatment of nerve degenerative diseases medicine - Google Patents
Sparassis crispa polyoses extract is preparing the purposes for the treatment of nerve degenerative diseases medicine Download PDFInfo
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- CN104958314B CN104958314B CN201510285844.XA CN201510285844A CN104958314B CN 104958314 B CN104958314 B CN 104958314B CN 201510285844 A CN201510285844 A CN 201510285844A CN 104958314 B CN104958314 B CN 104958314B
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- 241000272503 Sparassis radicata Species 0.000 title claims abstract description 35
- 239000000284 extract Substances 0.000 title claims abstract description 29
- 239000003814 drug Substances 0.000 title claims abstract description 9
- 210000005036 nerve Anatomy 0.000 title abstract description 6
- 208000015122 neurodegenerative disease Diseases 0.000 title abstract description 6
- 238000002360 preparation method Methods 0.000 claims abstract description 6
- 238000001291 vacuum drying Methods 0.000 claims abstract description 4
- 238000004321 preservation Methods 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 6
- 241000894006 Bacteria Species 0.000 claims description 3
- 239000012530 fluid Substances 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 2
- 208000018737 Parkinson disease Diseases 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- 239000013049 sediment Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 3
- 208000024827 Alzheimer disease Diseases 0.000 abstract description 2
- 206010039966 Senile dementia Diseases 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 abstract description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract 2
- 238000002474 experimental method Methods 0.000 abstract 1
- 238000000855 fermentation Methods 0.000 abstract 1
- 230000004151 fermentation Effects 0.000 abstract 1
- 238000000338 in vitro Methods 0.000 abstract 1
- 230000000324 neuroprotective effect Effects 0.000 abstract 1
- DIVDFFZHCJEHGG-UHFFFAOYSA-N oxidopamine Chemical compound NCCC1=CC(O)=C(O)C=C1O DIVDFFZHCJEHGG-UHFFFAOYSA-N 0.000 description 18
- 210000004027 cell Anatomy 0.000 description 17
- 230000006378 damage Effects 0.000 description 7
- 230000004112 neuroprotection Effects 0.000 description 7
- 230000004083 survival effect Effects 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 101710138657 Neurotoxin Proteins 0.000 description 3
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 210000003470 mitochondria Anatomy 0.000 description 3
- 210000001700 mitochondrial membrane Anatomy 0.000 description 3
- 239000002581 neurotoxin Substances 0.000 description 3
- 231100000618 neurotoxin Toxicity 0.000 description 3
- 239000002953 phosphate buffered saline Substances 0.000 description 3
- 230000008439 repair process Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 230000006907 apoptotic process Effects 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000004064 dysfunction Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000002438 mitochondrial effect Effects 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 206010006100 Bradykinesia Diseases 0.000 description 1
- 241001573498 Compacta Species 0.000 description 1
- 206010012289 Dementia Diseases 0.000 description 1
- 208000006083 Hypokinesia Diseases 0.000 description 1
- 206010044565 Tremor Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000007640 basal medium Substances 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 230000019771 cognition Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 230000000254 damaging effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000003412 degenerative effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 230000004771 dopaminergic neurodegeneration Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000008451 emotion Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007721 medicinal effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 210000003061 neural cell Anatomy 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 210000003523 substantia nigra Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides Sparassis crispa polyoses extract to prepare the purposes in treating nerve degenerative diseases medicine, is related to pharmaceutical technology field.Extracted with fermentation, ethanol, the Sparassis crispa polyoses extract of vacuum drying preparation, through experiment in vitro, it was demonstrated that there is the effect of significant neuroprotective activity.Preparation method of the present invention is simple, and cost is cheap, available for the medicine for preparing treatment nerve degenerative diseases, such as Parkinson's, senile dementia etc..
Description
Technical field
The invention discloses Sparassis crispa polyoses extract to prepare the purposes for the treatment of nerve degenerative diseases medicine, is related to doctor
Medicine technical field.
Background technology
Parkinson's (PD) are a kind of chronic CNS degenerative disorders.It can damage the limb function of patient,
Language ability etc., clinical manifestation are rest tremor, stiff, bradykinesia and posture unstability and cognition and the disturbance of emotion.
Its cause of disease at present it is still indefinite, but its major pathologic features be substantia nigra compacta in dopaminergic neuron loss and claimed
For the presence of the intracytoplasmic inclusion of Louis body, also the medicine of no obvious curative effects discloses use.
The content of the invention
The invention provides Sparassis crispa polyoses extract to prepare the purposes in treating nerve degenerative diseases medicine, has
Obvious medical effect.
The invention discloses the preparation method of the extract, it can be used for industrialized production.
The present invention further discloses medicinal application of the extract in terms of neuroprotection.
The Sparassis crispa mutagenic strain of the present invention：In China typical culture collection center preservation, preservation address：Wuhan Wuhan
University, preservation date：On May 3rd, 2015, specific name：Sparassis crispa SC031;Sparassis crispa SC031, preservation are stepped on
Mark is CCTCC NO:M2015275.
The preparation method of the extract with neuroprotection of the present invention is as described below：
The fermented tanks of Sparassis crispa SC301 are obtained into zymotic fluid, is centrifuged 5 ~ 10 minutes, abandoned with 4000 ~ 5000 revs/min
Clearly, precipitation is lyophilized, obtains Sparassis crispa agaric powder, and bacterium powder is extracted 2 times according to 100 times of amounts, 80 DEG C of hot water, and 3 hours every time, filtering was simultaneously
It is 1.15-1.20 to reclaim extract solution and be concentrated into room temperature relative density, vacuum drying, produces Sparassis crispa SC031 polyoses extracts.
The positive effect of the present invention is：Sparassis crispa SC031 polyoses extracts are provided in raising memory and improvement to recognize
Know the medical application of function, neuroprotection is obvious, to prevention and treatment Parkinson's and the effect of senile dementia also ten
Divide notable.
Brief description of the drawings
The influence of the survival rate for the DPC12 cells that Fig. 1 NNK0019 damage to 6-OHDA, cell is by NNK0019（10 μM,
20μM）3h is pre-processed, then adds 100 μM of 6-OHDA processing 24h.Data are presented with the percentage of control group, compared to control
Group ###p<0.001, compared to model group（100μM 6-OHDA）***p<0.001.
The repair for the DPC12 cells that Fig. 2 NNK0019 damage to 6-OHDA, cell is by NNK0019（20μM）Pre- place
3h is managed, then adds 100 μM of 6-OHDA processing 24h, control is blank group, and model is model group.
Embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention
Rather than the limitation scope of the invention.In addition, it is to be understood that after present disclosure has been read, those skilled in the art can be with
The present invention is made various changes or modifications, these equivalent form of values equally fall within the model that the application institute appended claims limits
Enclose.
The present invention points out a kind of Sparassis crispa polyoses extract, has obvious neuroprotection, can treat or prevent always
Applied in dementia and Parkinson medicinal or health products.
Embodiment 1：
The preparation of Sparassis crispa polyoses extract
The fermented tanks of Sparassis crispa SC031 are obtained into zymotic fluid, is centrifuged 5 ~ 10 minutes, abandoned with 4000 ~ 5000 revs/min
Clearly, precipitation is lyophilized, obtains Sparassis crispa agaric powder, and bacterium powder is extracted 2 times according to 100 times of amounts, 80 DEG C of hot water, and 3 hours every time, filtering was simultaneously
It is 1.15-1.20 to reclaim extract solution and be concentrated into room temperature relative density, vacuum drying, produces Sparassis crispa polyoses extract.
Embodiment 2：
The protective effect for the differentiated PC12 cells that Sparassis crispa SC031 polyoses extracts damage to neurotoxin 6-OHDA
The present invention passes through the result of mtt assay detection, the polyoses extract confrontation of reflection Sparassis crispa using cell survival rate as index
Neurotoxin 6-OHDA toxic action size, to illustrate its neuroprotection.
Take the logarithm the PC12 cells in growth period, 3*105/ml cell suspension is diluted to complete medium, per hole 100
μ l are inoculated in 96 orifice plates, are placed in 37 DEG C, 24h is cultivated in 5% CO2 incubators.Culture medium is removed, the Sparassis crispa for adding 100 μ l is more
Sugar extract solution（Final concentration 4 μ g/ml, 8 μ g/ml, prepared by basal medium as solvent）, model group and control group are distinguished
Add 100 μ l basal mediums, every group of 4 multiple holes.After medical preconditioning 3h, 6-OHDA is separately added into administration group and model group
（Final concentration of 100 μm of ol/l）, 24h is cultivated, the 5mg/ml μ l of MTT 10 is added, 4h is cultivated in 37 DEG C of incubators.Supernatant discarding
The μ l of DMSO 150 are added per hole afterwards, concussion is uniform, and each hole absorbance is determined at 540nm wavelength to reflect depositing for PC12 cells
Motility rate.
Above-mentioned MTT measurement results are shown（Fig. 1）, when only adding 6-OHDA, cell survival rate is decreased obviously compared with control group,
Illustrate that 6-OHDA has obvious damaging action to PC12 cells；, can be with when Sparassis crispa polyoses extract and 6-OHDA are incubated altogether
It is obvious to suppress the decline of cell survival rate, and strengthen with the increase of concentration, it is significant to illustrate that Sparassis crispa polyoses extract has
Neuroprotection.
6-OHDA damage PC12 cell models are to study the conventional neural cell model of Parkinson's, the chemistry point with reference to more than
Analysis, it is seen then that NNK0019 has neuroprotection, and has certain concentration dependent.
Embodiment 3：
The Sparassis crispa SC031 polyoses extracts sample PC12 cell lines to caused by neurotoxin 6-OHDA of above-described embodiment 1
The repair of the dysfunction of plastochondria
Molecular probe JC-1 can be used for determining mitochondrial membrane potential.JC-1 is a kind of lipophilic dye of positive ion, energy
It is selectively entered mitochondria.It can reversibly change (Cossarizza et from green to red when film potential raises
al. 1993).Healthy cell Mitochondria film potential is higher, and JC-1, which can be spontaneously formed, is referred to as J-aggregates with strong
Red fluorescence complex compound.On the other hand, apoptosis or unsound cell have relatively low mitochondrial membrane potential, and JC-1 is still
In monomeric, it has been only capable of displaying out green fluorescence (Cossarizza et al. 1993).
Take the logarithm the PC12 cells in growth period, 3*105/ml cell suspension is diluted to complete medium, per hole 1ml
6 orifice plates are inoculated in, 37 DEG C is placed in, 24h is cultivated in 5% CO2 incubators.Culture medium is removed, adds 1ml Sparassis crispa Polyose extraction
Thing（4 μ g/ml, 8 μ g/ml）, model group and control group are separately added into 1ml basal mediums.After medical preconditioning 3h, in administration group
6-OHDA is separately added into model group（Final concentration of 100 μm of ol/l）, cultivate 12h.10 points are handled at 37 DEG C with 2 μM of JC-1
Clock（ (Sigma-Aldrich, USA).After washing three times, phosphate buffered saline (PBS) is used（PBS）After washing three times, fluorescence is used
Fluorescence color change in microscope (Carl Ziess) observation mitochondria.
Above-mentioned mitochondrial membrane potential changes experimental result（Fig. 2）It has been shown that, only add the green fluorescence of 6-OHDA model group
Amount with red fluorescence ratio with control group compared with it is significantly raised, illustrate that 6-OHDA result in the damage of PC12 cell mitochondrials, cause
Its dysfunction.And when 6-OHDA and Sparassis crispa polyoses extract are acted on simultaneously, green red fluorescence ratio significantly reduces, and illustrates silk ball
Granulose extract, which has, repairs the function that 6-OHDA damages to PC12 cell mitochondrials, it is known that Sparassis crispa polyoses extract is to having
It may be by preventing it from further developing in apoptotic process, so as to play a protective role to nerve cell.
Claims (2)
1. Sparassis crispa SC301 polyoses extracts are preparing the application in treating Parkinson's disease medicament, described Sparassis crispa
SC301 is in China typical culture collection center preservation, preservation address：Wuhan Wuhan University, preservation date：May 3 in 2015
Day, specific name：Sparassis crispa SC301；Sparassis crispa SC301, preservation registration number are CCTCC NO:M2015275.
2. the preparation method of Sparassis crispa SC301 polyoses extracts as described in claim 1, it is prepared by following steps：
The fermented tanks of Sparassis crispa SC301 are obtained into zymotic fluid, is centrifuged 5 ~ 10 minutes with 4000 ~ 5000 revs/min, abandons supernatant, are sunk
Form sediment lyophilized, obtain Sparassis crispa agaric powder, bacterium powder extracts 2 times according to 100 times of amounts, 80 DEG C of hot water, 3 hours every time, filters and reclaims and carries
It is 1.15-1.20 to take liquid and be concentrated into room temperature relative density, vacuum drying, produces Sparassis crispa polyoses extract.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510285844.XA CN104958314B (en) | 2015-05-29 | 2015-05-29 | Sparassis crispa polyoses extract is preparing the purposes for the treatment of nerve degenerative diseases medicine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510285844.XA CN104958314B (en) | 2015-05-29 | 2015-05-29 | Sparassis crispa polyoses extract is preparing the purposes for the treatment of nerve degenerative diseases medicine |
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Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105384842A (en) * | 2015-12-17 | 2016-03-09 | 黑龙江众生生物工程有限公司 | Method for extracting water soluble beta-glucan from sparassis crispa sporophore |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007055952A (en) * | 2005-08-25 | 2007-03-08 | Unitika Ltd | Composition derived from sparassis crispa |
| KR20100032713A (en) * | 2008-09-18 | 2010-03-26 | (주)우성바이오 | Extract of sparassis crispa and its use as anti-inflammatory medicine |
-
2015
- 2015-05-29 CN CN201510285844.XA patent/CN104958314B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007055952A (en) * | 2005-08-25 | 2007-03-08 | Unitika Ltd | Composition derived from sparassis crispa |
| KR20100032713A (en) * | 2008-09-18 | 2010-03-26 | (주)우성바이오 | Extract of sparassis crispa and its use as anti-inflammatory medicine |
Non-Patent Citations (1)
| Title |
|---|
| 绣球菌多糖的提取工艺优化及其抗氧化作用;崔丽霞 等;《食品工业》;20131231;第34卷(第6期);摘要，材料与方法，第24页第2段 * |
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| CN104958314A (en) | 2015-10-07 |
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