CN105521075B - A kind of Rosa Damascana for treating Alzheimer's disease and its application - Google Patents
A kind of Rosa Damascana for treating Alzheimer's disease and its application Download PDFInfo
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- ASUTZQLVASHGKV-UHFFFAOYSA-N galanthamine hydrochloride Natural products O1C(=C23)C(OC)=CC=C2CN(C)CCC23C1CC(O)C=C2 ASUTZQLVASHGKV-UHFFFAOYSA-N 0.000 description 1
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- BUGYDGFZZOZRHP-UHFFFAOYSA-N memantine Chemical compound C1C(C2)CC3(C)CC1(C)CC2(N)C3 BUGYDGFZZOZRHP-UHFFFAOYSA-N 0.000 description 1
- 229960004640 memantine Drugs 0.000 description 1
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- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/738—Rosa (rose)
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
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- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to biomedicine field, specifically provides a kind of Rosa Damascana for treating Alzheimer's disease and its application.The Rosa Damascana is extracted from rose dried flower with steam distillation method and is prepared.The Rosa Damascana has significant therapeutic effect to Caenorhabditis elegans Alzheimer's disease pathological model, can be applied in the medicine for preparing treatment Alzheimer's disease, can also be applied in the relevant healthcare product of prevention and auxiliary treatment Alzheimer's disease are prepared.The preparation method of Rosa Damascana provided by the invention can be utilized the volatile oil component in rose dried flower, reduce and waste, economize on resources.
Description
Technical field
The invention belongs to biomedicine field, and in particular to a kind of Rosa Damascana for treating Alzheimer's disease and its should
With.
Background technology
Alzheimer's disease (Alzheimer'sM disease, AD) is a kind of with progressive cognitive disorder and memory damage
Nervous system degenerative disease based on wound, it is a kind of most common form of senile dementia.Clinically with memory disorders, mistake
The generalized dementia performance such as language, agnosia, the infringement of visual space technical ability, perform function obstacle and personality and behavior change is characterized.
Proportion is more than 75% in dementia patients.With the aggravation of China's aging degree, AD, which can turn into, influences senior health and fitness
One of principal disease, effectively preventing and treating AD turns into another key subjects of the pendulum in face of us.
Amyloid beta deposition (aβ protein) and neurofibrillary tangles are AD two big key pathological features.Modern medicine
Years of researches have been carried out to AD pathogenesis, but because its cause of disease is complicated, the sick pathogenesis is still indefinite so far,
" A β cascade hypothesis " is currently by one of well accepted mechanism of scientific circles.The theory thinks the A β of abnormal deposition in brain in patients
By a series of cascade reactions such as radical reaction, Mitochondrial oxidative damage and inflammatory reaction, god is directly or indirectly acted on
Through member and Deiter's cells, neuronal function exception or dead is ultimately resulted in, triggers cognitive impairment and amnesia, finally
Cause dementia.Amyloid precusor protein (APP) gene and presenilin genes are found in familial Alzheimer disease disease patient
Undergo mutation, cause A β to turn into theory strong evidence the most in the overexpression of brain.Therefore amyloid beta has turned into
The anti-AD drug targets of generally acknowledged screening.
The AD causes of disease are complicated, the course of disease is very long and morbidity link is more, it is necessary to Long-term taking medicine, anti-alzheimer ' main at present
Silent medicine acetylcholinesteraseinhibitors inhibitors (such as galanthamine) and N-methyl-D-aspartate acceptor (nmda receptor) antagonist
(such as Memantine), but said medicine is expensive, illusion after medication, misunderstanding, dizziness, headache and the side effect such as tired show
Write, and can only delay, but can not control or reverse conditions of patients.
And Chinese medicine composition has various active component, the feature of Mutiple Targets, Western medicine can be overcome not to a certain extent
Foot part so that screen from Chinese medicine composition treasure-house and had a extensive future for treating AD drug candidates.
Caenorhabditis elegans (Caenorhabditis elegans) be a highly useful multifunctional drug screening and
The research platform of mechanism of drug action.Its cheap, easily culture；Generation cycle is short, offspring's number is more, while can obtain a large amount of backgrounds
Consistent individual, ensure experimental repeatability and tested using large sample size, so as to exclude the influence of individual difference；With it is high
Biology is highly conserved in terms of gene and molecular pathway (Kaletta and Hengartner, 2006).It is as one from thin
Born of the same parents' levels in vitro medicine is just sieved to a bridge of horizontal medicine secondary screening in Mice Body, is increasingly favored by pharmacy man.It is beautiful
DevGen companies of state have entered clinical research by FDA approvals using the arrhythmia medicine of humanization nematode screening.
The foundation of humanization Caenorhabditis elegans AD pathological models is that people source A β 1-42 genes are transferred into myosin to start
Sub- downstream and expression temperature controlled, the nematode strain can be in 15 DEG C of normal growths, the A β 1-42 tables under the conditions of being transferred to 25 DEG C
Reach and accumulated in its intramuscular, cause nematode to lose locomitivity and benumb.Muscular paralysis phenotype can be delayed by adding test medicine,
The percentage that its result accounts for tested nematode population quantity with muscular paralysis nematode population represents.It is higher that nematode ratio is not benumbed, medicine
The anti-AD effects of thing are more notable.This model early has been used for anti-AD diseases mechanism and anti-AD medicines and candidate mechanism of action
Research (Link, 1995；2001).
Rose is the Chinese medicine of the promoting flow of qi and blood circulation simply, and big geneva leather rose chloroform extract is proved to that in vitro culture can be promoted
Rat embryonic neuronal cell axon growth, and suppress neuron axon atrophy caused by A β (25-35) (Awale et al.,
2011).Rosa Damascana has anti-AD activity, and further studies have shown that Rosa Damascana moderate resistance AD main active is benzene second
Alcohol (C8H10O), it, which has, suppresses activity of cholinesterase and BuCh lipase activity (Senol et al., 2011).City at present
The Rosa Damascana sold is extracted from new win in rose mostly, and the fresh rose for failing to utilize in time is needed using the side such as high salt
Method suitably preserves, and just can guarantee that the quality of extraction gained essential oil.And Rosa Damascana is extracted from rose dried flower and is used to treat A Er
There is not been reported for Ci Haimo diseases.
Based on the studies above present situation, the present invention now provides a kind of Rosa Damascana for treating Alzheimer's disease, the rose
Essential oil is that the volatile oil obtained is extracted from rose dried flower, and the Rosa Damascana is to Caenorhabditis elegans Alzheimer's disease pathology mould
Type has significant therapeutic effect.Present invention simultaneously provides the application of the Rosa Damascana.
Bibliography
Hou Y,Wang Y,Zhao J,Li X,Cui J,et al.(2014)Smart soup,a traditional
Chinese medicine formula,ameliorates amyloid pathology and related cognitive
deficits.PLoS ONE 9(11):e111215.doi:10.1371/journal.pone.0111215.
Kaletta T,Hengartner MO.Finding function in novel targets:C.elegans
as a model organism.Nat Rev Drug Discov 2006,5:387e398.
Link CD.Expression of human beta-amyloid peptide in transgenic
Caenorhabditis elegans.Proc Natl Acad Sci USA 1995,92:9368e9372.
Link CD.Transgenic invertebrate models of age-associated
neurodegenerative diseases.Mech Ageing Dev 2001,122:1639e1649.
Awale S,Tohda C,Tezuka T et al.Protective Effects of Rosa damascena
and its active constituent on Aβ(25–35)-induced neuritic atrophy.Evidence-
Based Complementary and Alternative Medicine 2011,doi:10.1093/ecam/nep149.
Senol FS,Orhan I,Kurkcuoglu M,et al.An in vitro approach to
neuroprotective activity of Rosa damascena Mill,a medieval age traditional
medicine used for memory enhancement.Planta Med.2011,77(12):1440-1440.
Around, Zhou little Ping, State of Zhao is grand, Liu Hongwei, Ding Lan, the research colors of Chen Li benevolence China bitter water rose oil fragrance component
Compose 2002,20 (6):560-564.
The content of the invention
It is an object of the present invention to provide a kind of Rosa Damascana for treating Alzheimer's disease, the Rosa Damascana has treatment A Er
The potentiality of Ci Haimo diseases.
It is a further object to provide the application of above-mentioned Rosa Damascana.
Above-mentioned purpose of the present invention is to be achieved through the following technical solutions：
A kind of Rosa Damascana for treating Alzheimer's disease, the Rosa Damascana are prepared with steam distillation method：Rose is taken to do
The weight ratio of flower, addition distilled water, rose dried flower and distilled water is 1:10, steam distillation extraction 4h, collect the rose distillated
Essential oil, the Rosa Damascana distillated afterwards with ether extraction, after ether evaporates into no ether taste, collect Rosa Damascana.
The rose is bitter water rose (Rosa Sertata x Rosa rugosa).
A kind of application of Rosa Damascana in the medicine for preparing treatment Alzheimer's disease.
The Rosa Damascana is capsule, tablet, granule, powder, oral liquid, ball as the pharmaceutical preparation of effective component
Agent.
A kind of application of Rosa Damascana in the health products of prevention and auxiliary treatment Alzheimer's disease are prepared.
The Rosa Damascana is capsule, tablet, granule, powder, oral liquid, ball as the health products formulation of active ingredient
Agent.
The beneficial effects of the present invention are：
1st, the present invention is tested using Caenorhabditis elegans as Alzheimer's disease pathological model, the results showed that：This hair
The Rosa Damascana of bright offer has significant therapeutic effect to the Caenorhabditis elegans for suffering from Alzheimer's disease.It can be controlled in preparation
Treat and applied in the medicine of Alzheimer's disease, can also be applied in the health products for preparing prevention Alzheimer's disease.
2nd, the preparation method of Rosa Damascana of the present invention is to extract volatile oil from rose dried flower, with presently commercially available rose
Rare essential oil is compared, and the Rosa Damascana has lost Rosa Damascana characteristic chemical constituent citronellol and geraniol, but to beautiful hidden bar
The therapeutic effect of nematode AD pathological models is constant.This preparation method can make the volatile oil component in rose dried flower obtain profit
With reduction wastes, and economizes on resources.
Technical scheme is described in further detail with reference to specific embodiment, but protection scope of the present invention is not
It is confined to as described below.
Brief description of the drawings
Therapeutic action of Fig. 1 Rosa Damascanas to Caenorhabditis elegans AD pathological models
Control- blank control groups, H- high concentration fresh rose flower essential oil groups, concentration fresh rose flower essential oil group in M-, L- are low
Concentration fresh rose flower essential oil group；HH- high concentration rose dried flower essential oil groups, concentration rose dried flower essential oil group in MM-, LL- low concentrations
Rose dried flower essential oil group
Embodiment
The preparation method of 1 Rosa Damascana of the present invention of embodiment
The extraction of fresh rose flower essential oil：100g fresh rose flowers are taken, add 1000mL distilled water, steam distillation extracts 4h,
The μ L of Rosa Damascana 30 distillated are collected, the Rosa Damascana distillated afterwards with ether extraction, after ether evaporates into no ether taste,
Rosa Damascana is collected, it is final to obtain the μ L of Rosa Damascana 20.
The extraction of rose dried flower essential oil：100g rose dried flowers are taken, add 1000mL distilled water, steam distillation extracts 4h,
The μ L of Rosa Damascana 15 distillated are collected, the Rosa Damascana distillated afterwards with ether extraction, after ether evaporates into no ether taste,
Rosa Damascana is collected, it is final to obtain the μ L of Rosa Damascana 10.
The GC/MS testing results of 2 Rosa Damascana of the present invention of embodiment
GC/MS detects above-mentioned Rosa Damascana composition and the results are shown in Table 1 and table 2.
The Rosa Damascana composition analysis result extracted in the fresh rose flower of table 1
The Rosa Damascana composition analysis result extracted in the rose dried flower of table 2
Above-mentioned GC/MS testing results show：Main component is not in the Rosa Damascana extracted in rose dried flower and fresh rose flower
Together, the Rosa Damascana extracted in rose dried flower has lost Rosa Damascana characteristic chemical constituent citronellol and geraniol.
The anti-AD of Rosa Damascana extracted in the rose dried flower of embodiment 3 Effect study
1st, biomaterial
(1) Caenorhabditis elegans CL4176 is purchased from Caenorhabditis Genetics Center (CGC)；For transgenosis
Strain, in 25 DEG C of temperature-induced lower muscle specific expression mankind A β1-42, A β1-42Assemble in musculature, ultimately result in nematode
Paralysis, the present embodiment is using C. Elegans Automatic Screening strain CL4176 as the pathological model for screening anti-AD medicines.
(2) Escherichia coli OP50 (uracil leaky mutant), purchased from Caenorhabditis Genetics Center
(CGC), the food as Caenorhabditis elegans.
2nd, reagent
(1) solid NGM (Nematode Growth Medium) medium components are with making (being upgraded to example with 1)：
After solid NGM culture mediums prepare, high-pressure constant temp sterilizing 20min, 5mg/ is added under aseptic operating platform at 121 DEG C
ML cholesterol 1mL, 1M MgSO4 1mL,1M CaCl21mL shakes up, and pours into sterilized 9cm culture plates, about 20mL/ plates while hot.
Stand and wait culture medium solidifying, it is standby.
(2) M9 formula of liquid
| Composition | Content |
| Na2HPO4 | 6.00g |
| KH2PO4 | 3.00g |
| NaCl | 5.00g |
| 1M MgSO4 | 1.00mL |
| Supplement H2O is extremely | 1000mL |
(3) preparation of lysate：6.4%NaClO solution and 1M NaOH solutions by volume 1:1 mixing.
3rd, implementation steps
(1) culture of nematode：
Nematode is connected on the solid NGM plates for scribbling Escherichia coli OP50, is subsequently placed in 16 DEG C of incubator and cultivates, when
Synchronizing is handled when nematode grows to adult.
(2) nematode synchronizes：
Select containing a large amount of adults and the NGM culture mediums that there are part line eggs to hatch, with M9 liquid by nematode from training
Support and swept away on base, is transferred in centrifuge tube, standing makes nematode free settling abandon supernatant to ttom of pipe.How much sight worm amount is to centrifugation
Lysate is added in pipe, stops being vortexed when vibrating 5-7 minutes when nematode complete rupture on whirlpool misfortune agitator, and be sub-packed in
In 1.5mL centrifuge tubes, line eggs are washed three times with M9 solution.
(3) in rose dried flower Rosa Damascana preparation
The extraction of fresh rose flower essential oil：100g fresh rose flowers are taken, add 1000mL distilled water, steam distillation extracts 4h,
The μ L of Rosa Damascana 30 distillated are collected, the Rosa Damascana distillated afterwards with ether extraction, after ether evaporates into no ether taste,
Rosa Damascana is collected, it is final to obtain the μ L of Rosa Damascana 20.
The extraction of rose dried flower essential oil：100g rose dried flowers are taken, add 1000mL distilled water, steam distillation extracts 4h,
The μ L of Rosa Damascana 15 distillated are collected, the Rosa Damascana distillated afterwards with ether extraction, after ether evaporates into no ether taste,
Rosa Damascana is collected, it is final to obtain the μ L of Rosa Damascana 10.
(4) experimental group is set
Blank control group (Control)：0.1%DMSO (v/v).
High concentration fresh rose flower essential oil group (H)：The Rosa Damascana of 5000 times of dilutions, solvent 0.1%DMSO.
Middle concentration fresh rose flower essential oil group (M)：The Rosa Damascana of 10000 times of dilutions, solvent 0.1%DMSO.
Low concentration fresh rose flower essential oil group (L)：The Rosa Damascana of 50000 times of dilutions, solvent 0.1%DMSO.
High concentration rose dried flower essential oil group (HH)：The Rosa Damascana of 5000 times of dilutions, solvent 0.1%DMSO.
Middle concentration rose dried flower essential oil group (MM)：The Rosa Damascana of 10000 times of dilutions, solvent 0.1%DMSO.
Low concentration rose dried flower essential oil group (LL)：The Rosa Damascana of 50000 times of dilutions, solvent 0.1%DMSO.
(4) experimental procedure
A. the preparation of the NGM culture mediums containing Rosa Damascana：Each group Rosa Damascana is taken, isometric sky with Rosa Damascana group
White control group, it is separately added into NGM culture mediums, the final concentration of each group Rosa Damascana is stood etc. respectively as described in being set experimental group
Treat culture medium solidifying.Foods of the even spread Escherichia coli OP50 as nematode on culture medium.
B. the line eggs being sub-packed in after synchronization in centrifuge tube are transferred to respectively and are coated with OP50 and are mixed with different dense
In the NGM culture dishes of the Rosa Damascana of degree, blank control group is be coated with OP50 contain with Rosa Damascana same volume 0.1%
DMSO NGM culture dishes, each 60 nematodes of culture dish, each drug concentration setting three is parallel, and 16 DEG C are cultivated 3 days to L3
Phase.
In order that nematode expresses A β, L3 phase nematodes are placed at 25 DEG C and induced, nematode paralysis is started counting up after 34h
Bar number.Every two hours count once, until all nematodes are all benumbed.When paralysis refers to mechanical stimulus nematode body, nematode can not
Motion or only head movement.
(5) experimental result
The Rosa Damascana of table 3 delays AD nematode half paralysis times TD50 to influence
| Group | Half paralysis time (h) |
| C | 39.2±0.15a |
| L | 41.5±0.09b |
| M | 41.3±0.11b |
| H | 42.7±0.58c |
| LL | 41.3±0.11b |
| MM | 41.9±1.33c |
| HH | 43.4±0.07c |
Note：Different letters represent significant difference (p<0.05)
Fig. 1 ordinate represents the percentage accounted for without the Caenorhabditis elegans of paralysis, and in point at the same time, this value is got over
Greatly, show not have in the treatment group Caenorhabditis elegans ratio of paralysis bigger, i.e., medicine delays the effect of paralysis stronger.
Test result indicates that Rosa Damascana provided by the invention has significant retarding action to AD nematode muscular paralysises,
This prompting Rosa Damascana has anti-AD effects (Fig. 1), wherein, the half paralysis time of middle concentration rose dried flower essential oil group (MM) shows
Work is better than middle concentration fresh rose flower essential oil group (M) (table 3).
Proved by above example, Rosa Damascana provided by the invention is to Caenorhabditis elegans Alzheimer's disease pathology
Model has significant therapeutic effect, prompts Rosa Damascana of the present invention to have the potentiality for the treatment of Alzheimer's disease, can be
Prepare and applied in the medicine for the treatment of Alzheimer's disease, can also prepare prevention guarantor related to auxiliary treatment Alzheimer's disease
Applied in strong product.
Claims (3)
- A kind of 1. Rosa Damascana for treating Alzheimer's disease, it is characterised in that the Rosa Damascana extracts from bitter water rose, Rosa Damascana is prepared with steam distillation method：Take rose dried flower, add distilled water, the weight ratio of rose dried flower and distilled water is 1: 10, steam distillation extraction 4h, the Rosa Damascana distillated is collected, the Rosa Damascana distillated afterwards with ether extraction, treats that ether is waved After being sent to no ether taste, Rosa Damascana is collected.
- A kind of 2. application of Rosa Damascana as claimed in claim 1 in the medicine for preparing treatment Alzheimer's disease.
- 3. application as claimed in claim 2, it is characterised in that the Rosa Damascana is capsule as the pharmaceutical preparation of effective component Agent, tablet, granule, powder, oral liquid, pill.
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