CN107308199A - The application of Japanese Raspberry Root or its extract in fat-reducing medicament is prepared - Google Patents
The application of Japanese Raspberry Root or its extract in fat-reducing medicament is prepared Download PDFInfo
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- CN107308199A CN107308199A CN201710570325.7A CN201710570325A CN107308199A CN 107308199 A CN107308199 A CN 107308199A CN 201710570325 A CN201710570325 A CN 201710570325A CN 107308199 A CN107308199 A CN 107308199A
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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)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
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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
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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Abstract
The present invention relates to medicinal usage technical field, the more particularly to application of Japanese Raspberry Root or its extract in fat-reducing medicament is prepared.In gene level, result of study of the present invention shows that Japanese Raspberry Root can raise LDLR genes with conspicuousness, and conspicuousness lowers MTP genes, APOB genes and DGAT1 genes；And the effect for reducing fat of Japanese Raspberry Root is better than affine cudweed, two kinds of congeners of spoon leaf affine cudweed.In cellular level, as a result show that Japanese Raspberry Root can promote cell to LDL absorbability, that is, promote the removing of LDL in the circulatory system；The content of the ester of intracellular glycerol three can be reduced.As a result show that Japanese Raspberry Root or its extract have significant effect for reducing fat, mechanism of action is clear and definite, and Japanese Raspberry Root cost is low, convenient sources, prepares simply, has no toxic side effect.
Description
Technical field
The present invention relates to medicinal usage technical field, more particularly to Japanese Raspberry Root or its extract is in fat-reducing medicament is prepared
Application.
Background technology
Hyperlipidemia refers to that blood lipid level is too high, refers specifically to blood cholesterol (TC, LDL-c) and/or triglycerides (TG)
Too high or HDL-C (HDL-C) is too low.Clinical manifestation is mainly lipid and caused Huang is deposited in intradermal
The artery sclerosis of color knurl and lipid in the blood vessels caused by skin deposition.It is strong that hyperlipidemia can directly cause some to seriously endanger human body
The disease of health, such as atherosclerosis, coronary heart disease, pancreatitis, annual 30000000 people in the whole world die from phase caused by high fat of blood
Related disorders, it is the most serious for cardiovascular and cerebrovascular disease.With the improvement of living standards, the incidence of disease of China's coronary heart disease, the death rate
Also substantially rise, hyperpietic adds 25% in 10 years, at present up to 200,000,000 people or so.Counted according to the Ministry of Public Health, China's heart and brain
Vascular disease's First episode age 1/3, the incidence of disease was up to 13.6%, old man's cardiovascular and cerebrovascular of over-65s below 60 years old
Disease incidence is up to 95%, and all different degrees of presence is nervous, uncomfortable in chest, dizzy, high blood pressure, high, tired blood fat, hemiplegia, the heart are twisted
The symptoms such as pain, the incidence of disease is high.Daily with the speed increase of 20,000 people, the 40.72% of total death toll is accounted for.According to the World Health Organization
(WHO) count：Cardiovascular and cerebrovascular disease number is died from up to 17,500,000 people in the whole world in 2012.Cardiovascular and cerebrovascular disease turns into human health
" the first killer ".To improve prevention, treatment ability of medicine and the Scientific research level to cardiovascular and cerebrovascular disease, fat-reducing medicament is engaged in
Research is the problem of people compare concern in recent years.
Current clinically most widely used class lipid-lowering medicine, i.e. 3-Hydroxy-3-methylglutaryl CoA A (HMG-CoA) are also
Reductase inhibitor, i.e. statins.The conventional statins of clinic has Lovastatin (Lovastatin), Simvastatin
(Simvastatin), Pravastatin (Pravastatin), Fluvastatin (Fluvastatin), Atorvastatin
(Atorastatin).Such medicine can suppress the activity of the rate-limiting enzyme of intracellular cholesteryl synthesis early stage, cause intracellular
Free cholesterol is reduced, and raises the expression of cell surface LDL receptor (LDLR) by feedback, accelerates circulation
The removing of low-density lipoprotein (LDL-c) in blood, beneficial to the removing and transhipment of T-CHOL (TC).Except LDLR target drugs,
There are some with microsomal triglyceride transfer protein (MTP also makees MTTP) and apolipoprotein B100 (APOB100) for target in recent years
The new lipid-regulation medicine of point goes through clinical practice.Lome Tapai (lomitapide, trade name Juxtapid) suppresses as MTP
Agent was ratified to list in 2012 by U.S. FDA, and FDA approvals meter Bo Mei in 2013 gives birth to (Mipomersen) and is used as APOB100Second
It can combine for ASON individually or with other lipid-regulation medicines for treating homozygous familial hypercholesterolemia
(HoFH).Main reduction TG medicine has three kinds：Peroxisome proliferator-activated receptor alpha (PPAR α) and activation lipoprotein
Enzyme LPL fibrate, nicotinic acid class and high-purity fish oil preparation, wherein nicotinic acid class medicine are gradually faded out pharmaceutical market.But on
State these medicines and be all inevitably present side effect.Side effect has headache, insomnia, the change of transience bowel habit, nausea etc.,
Occur more in prolonged application, it is short-term rare.Such medicine can produce more serious side effect, the lighter after being shared with Gemfibrozil Capsules
There is rhabdomyolysis, melanuria, kidney function damage etc. in hepatic disorder, DOMS, severe one.While the medicine target of chemicals
Point is single, it is often necessary to which composite reagent is to obtain more preferable therapeutic effect.
Reducing Blood Lipid by Chinese medicine has the advantages that mechanism of action uniqueness, lasting medicine, Mutiple Targets, multipath, toxic side effect are small, right
There are huge potentiality and advantage in reducing blood lipid.Japanese Raspberry Root (Gnaphalium_japonicum Thunb) is feverfew mouse
The herb of the Japanese affine cudweed of Anacamptis pyromaidalis category, also known as spire affine cudweed, clear and bright grass etc., nature and flavor are sweet, cool, nontoxic, return lung, liver, the spleen channel.Tool
Have the function of inducing diaphoresis, heat-clearing, improving eyesight, diuresis, available for curing cold, cough, have a headache, having sore throat, cataract, urine heat closes, drenches
It is turbid, leukorrhea, furunculosis etc..Modern pharmacology research shows the effects such as Japanese Raspberry Root has anti-inflammatory, treatment diarrhoea, carbuncle swells.Existing skill
The compound containing Japanese Raspberry Root is paid close attention to art is used for clear liver lung heat, relieving cough and reducing sputum, removing toxic substances diuresis, the function of anti-inflammation more.Dong Liang
" a kind of Soboring-up liver-protecting health protection tea and its preparation side of clear et al. " a kind of Sobering-up tea drink and preparation method thereof " and Huanglong et al.
Purposes of two kinds of compounds containing Japanese Raspberry Root on the antiemetic that relieves the effect of alcohol, protection liver is disclosed in method "." the one of Gao Peihuan et al.
A kind of blood-fat reducing composition containing potentilla discolor is disclosed in the composition of kind reduction blood fat ", it is mentioned that potentilla discolor also known as day
Blue or green ground is white, and potentilla discolor (Potentilla discolor Bge) is the herb of rosaceous plant potentilla discolor, reddish black with composite family
Ground is not same Chinese medicine in vain." gnaphalium affine total flavonoid improves and made Cui Jue et al. paper to diabetic mice metabolism disorder of blood lipid
Research ", it was recently reported that the drop blood of Japanese Raspberry Root grass congener affine cudweed (also known as affine cudweed, Gnaphalium offine)
Fat function, affine cudweed and Japanese Raspberry Root grass belong to composite family Gnaphalium, but are not of the same race, and the two is on meridian distribution of property and flavor and work(
There is difference in effect, affine cudweed nature and flavor are sweet, flat, return lung warp, affine cudweed is relieving cough and asthma, hypotensive, wind-damp dispelling, for catching a cold
Cough, bronchitis, asthma, hypertension, favism, lumbago, skelalgia of rheumatism；Traumatic injury, venomous snake bite, without heat-clearing are controlled in external application
Effect of removing toxic substances, it can be seen that even if two kinds of plants are all Gnaphaliums, but clinical application emphasis is different, thus speculates Buddhist ear
The lipid-reducing function of grass can not replace the lipid-reducing function of Japanese Raspberry Root.Therefore Japanese Raspberry Root simple and compound medicine are there is no at present
Relevant report with lipid-reducing function.
The content of the invention
In view of this, the application the invention provides Japanese Raspberry Root or its extract in fat-reducing medicament is prepared.The present invention
Research shows that Japanese Raspberry Root or its extract have significant effect for reducing fat, and mechanism of action clearly, has no toxic side effect.
In order to realize foregoing invention purpose, the present invention provides following technical scheme：
The invention provides the application of Japanese Raspberry Root or its extract in fat-reducing medicament is prepared.
The effect for reducing fat for the Japanese Raspberry Root that the present invention is confirmed, is specifically included：
1) effect of the Japanese Raspberry Root to lipid-loweringing key gene is confirmed using qPCR method：Japanese Raspberry Root can be with conspicuousness
LDLR genes are adjusted, conspicuousness lowers MTP genes, APOB genes and DGAT1 genes；
2) confirm that the effect for reducing fat of Japanese Raspberry Root is better than affine cudweed, two kinds of spoon leaf affine cudweed and belongs to plant together using qPCR method
Thing.Particularly Japanese Raspberry Root can raise LDLR genes, and other two kinds of medicinal materials lower LDLR genes；
3) effect for reducing fat of Japanese Raspberry Root is confirmed using cytologic experiment：Streaming result shows that Japanese Raspberry Root can promote carefully
Born of the same parents promote the removing of LDL in the circulatory system to LDL absorbability；Intracellular is determined with GPO method
Three ester levels show that Japanese Raspberry Root can reduce the content of the ester of intracellular glycerol three.
In the present invention, fat-reducing medicament is T-CHOL, triglycerides or LDL-C in reduction serum
Middle one or more of medicine.
Preferably, Japanese Raspberry Root extract is Japanese Raspberry Root alcohol extract.
Preferably, Japanese Raspberry Root extract is Japanese Raspberry Root ethanol extract.
Preferably, Japanese Raspberry Root extract is made using soxhlet extraction.
Preferably, the preparation method of Japanese Raspberry Root extract is：It is in terms of g/mL, Japanese Raspberry Root herb and ethanol is water-soluble
Liquid presses 1：The ratio mixing of (10~20), extracts 2~4h under the conditions of 80~90 DEG C, and filtering retains filtrate, concentrated, dries.
In the embodiment that the present invention is provided, the preparation method of Japanese Raspberry Root extract is：In terms of g/mL, by Japanese Raspberry Root
Herb and 90% ethanol press 1：15 ratio mixing, 3h is extracted under the conditions of 85 DEG C, is filtered, retains filtrate, is concentrated, is dried.
Preferably, the formulation of fat-reducing medicament is oral formulations or ejection preparation.
Preferably, oral formulations are tablet, pill, granule, oral liquid or capsule.
Preferably, ejection preparation is injection or injection powder injection.
Preferably, fat-reducing medicament also includes other fat-reducing medicaments.
The invention provides the application of Japanese Raspberry Root or its extract in fat-reducing medicament is prepared.The technology that the present invention has
Effect is：
In gene level, result of study of the present invention shows that Japanese Raspberry Root can be raised under LDLR genes, conspicuousness with conspicuousness
Adjust MTP genes, APOB genes and DGAT1 genes；And the effect for reducing fat of Japanese Raspberry Root is better than affine cudweed, two kinds of spoon leaf affine cudweed together
Platymiscium.
In cellular level, as a result show that Japanese Raspberry Root can promote cell to LDL absorbability, that is, promote the circulatory system
Middle LDL removing；The content of the ester of intracellular glycerol three can be reduced.
The above results show Japanese Raspberry Root or its extract has significant effect for reducing fat, and mechanism of action is clear and definite and reddish black
The white cost in ground is low, convenient sources, prepares simply, has no toxic side effect.
Brief description of the drawings
Fig. 1 shows regulating and controlling effect of the Japanese Raspberry Root extract to LDLR, MTP, APOB and DGAT1 lipid metabolism related gene, its
In, * p<0.05, * * p<0.01；
Fig. 2 shows that Japanese Raspberry Root extract, affine cudweed, spoon leaf affine cudweed are related to DGAT1 lipid metabolis to LDLR, MTP, APOB
The regulating and controlling effect of gene；
Fig. 3 shows intracellular DiI-LDL contents, wherein, * p<0.05；
Fig. 4 shows the content of intracellular triglyceride, wherein, * p<0.05.
Embodiment
The invention discloses the application of Japanese Raspberry Root or its extract in fat-reducing medicament is prepared, those skilled in the art can
To use for reference present disclosure, technological parameter realization is suitably modified.In particular, all similar replacements and change are to this
It is it will be apparent that they are considered as being included in the present invention for art personnel.The method of the present invention and application are
It is described by preferred embodiment, related personnel substantially can be not departing from present invention, in spirit and scope to herein
Described methods and applications are modified or suitably change is with combining, to realize and apply the technology of the present invention.
In following examples, the extract of Japanese Raspberry Root extract is Japanese Raspberry Root ethanol extract.Detailed process is：Claim
Japanese Raspberry Root herb 10g, plus the ethanol of 150mL 90% are taken, 85 DEG C are heated to, 3h is extracted, filtering retains filtrate, concentrated, dries,
Obtain extract medicinal extract.
The Japanese Raspberry Root or its extract that the present invention is provided agents useful for same or instrument in the application in preparing fat-reducing medicament
It can be bought by market.
With reference to embodiment, the present invention is expanded on further：
Embodiment 1
The embodiment of the present invention detects that Japanese Raspberry Root is related to lipid metaboli using fluorescence real-time quantitative PCR technology (RT-qPCR)
The influence of gene, it is determined that Japanese Raspberry Root has effect for reducing fat.Wherein, the gene paid close attention to has LDLR, MTP, APOB (to refer in particular to
The APOB of expression in liver100) and DGAT1.
Gene LDLR, LDL receptor is positioned on liver plasma membrane, passes through LDL in endocytosis mediate blood
Removing, more than 75% LDL is removed by LDLR in blood.
Triglycerides, cholesteryl ester and phospholipid are transported to newly-generated by MTP, microsome triacylglycerol transport protein
On APOB, VLDL particle (VLDL) is assembled into.
APOB, apolipoprotein B is produced by liver, is newborn VLDL proprietary component, and VLDL is assembled in liver cell, point
Secrete after circulation through serial degreasing, apoenzyme formation LDL.Therefore, it can promote LDL removing by raising LDLR, suppress MTP
With APOB expression, VLDL secretion and the conversion to LDL can be reduced.
DGAT1 is the key enzyme of triglycerides (TG) synthesis, catalyzes and synthesizes the final step of TG synthesis, DGAT1 defects
Mouse increases the sensitiveness of insulin and leptin and can resist the obesity of diet induced.
Present invention discover that Japanese Raspberry Root can raise LDLR, the expression of MTP and APOB genes is lowered, so as to reduce low-density
Lipoprotein cholesterol level.DGAT1 expression is lowered simultaneously, reduces the synthesis of triglycerides.
The present invention utilizes the expression of RT-qPCR method validation said genes, and its process is：
1st, control DMSO and Japanese Raspberry Root extract (100 μ g/mL) handle cell 24 hours.
2nd, extract RNA using kit and be reversed to cDNA.
3rd, RT-qPCR reacts, program：95 DEG C, 3min, (95 DEG C, 3s, 60 DEG C, 30s) 40 circulations.
Primer sequence is：
LDLR：Sense primer ACCAACGAATGCTTGGACAAC,
Anti-sense primer ACAGGCACTCGTAGCCGAT；
MTP：Sense primer ACAAGCTCACGTACTCCACTG,
Anti-sense primer TCCTCCATAGTAAGGCCACATC；
APOB：Sense primer ACACACTGGACGCTAAGAGGA,
Anti-sense primer ACTTGTGCTACCATCCCATACT；
DGAT1：Sense primer TATTGCGGCCAATGTCTTTGC
Anti-sense primer CACTGGAGTGATAGACTCAACCA.
3 experimental results of qPCR are shown in Table 1 and Fig. 1：
Change multiple of the Japanese Raspberry Root extract of table 1 to lipid metabolism related gene
Note：Change multiple refers to that drug-treated group influences gene variation multiple than DMSO control group.
QPCR results show that Japanese Raspberry Root can raise LDLR genes with conspicuousness, and conspicuousness lowers MTP genes, APOB genes
With DGAT1 genes.
Embodiment 2
The embodiment of the present invention compares Japanese Raspberry Root (Gnaphalium japonicum) and other two kinds of congeners
Affine cudweed (Gnaphalium offine), spoon leaf affine cudweed (Gnaphalium pensylvanicum) are to the work of said gene
With test method be the same as Example 1.Wherein, affine cudweed extract, the extracting method and Japanese Raspberry Root extracting method of spoon leaf affine cudweed
It is identical, it is alcohol extracting thing, working concentration is 100 μ g/mL.QPCR results (Fig. 2) show that the effect of Japanese Raspberry Root is significantly stronger than
Other two kinds of medicinal materials, particularly Japanese Raspberry Root can raise LDLR genes, and other two kinds of medicinal materials lower LDLR genes, therefore
Present invention demonstrates that the effect for reducing fat of Japanese Raspberry Root is better than affine cudweed and spoon leaf affine cudweed.
Embodiment 3
The embodiment of the present invention verifies the effect of Japanese Raspberry Root using the absorption for detecting fluorescence labeling low-density lipoprotein, its
Process is：
1st, control DMSO and Japanese Raspberry Root extract (100 μ g/mL) handle cell 24 hours.
2nd, the culture medium containing medicine is discarded, PBS rinsings cell twice, adds the DMEM culture mediums without serum, simultaneously
DiI-LDL (20 μ g/mL) is added to continue to be incubated 4 hours；
3rd, collected by trypsinisation cell, the intracellular DiI-LDL contents (emission of flow cytometer detection:541nm；
excitation:575nm)。
Streaming result is shown in Fig. 3, as a result shows that Japanese Raspberry Root can promote cell to LDL absorbability, that is, promotes circulation
LDL removing in system.
Embodiment 4
The embodiment of the present invention detects intracellular content of triglyceride using triglyceride reagent box, checking Japanese Raspberry Root
Act on, its process is：
1st, enuatrol (60 μ g/mL) induction HepG2 cells 24 hours；
2nd, control DMSO and Japanese Raspberry Root extract (100 μ g/mL) continue with cell 24 hours；
3rd, the content of triglycerides in cell lysis, kit detection cell.
Result of the test is shown in Fig. 4, as a result shows that Japanese Raspberry Root can reduce the content of the ester of intracellular glycerol three.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. the application of Japanese Raspberry Root or its extract in fat-reducing medicament is prepared.
2. application according to claim 1, it is characterised in that the fat-reducing medicament is T-CHOL in reduction serum, sweet
One or more of medicine in oily three esters or low-density lipoprotein.
3. application according to claim 1 or 2, it is characterised in that the Japanese Raspberry Root extract is Japanese Raspberry Root alcohol extracting
Thing.
4. application according to claim 3, it is characterised in that the Japanese Raspberry Root extract uses surname extraction legal system
.
5. application according to claim 4, it is characterised in that its preparation method is：In terms of g/mL, by Japanese Raspberry Root herb
1 is pressed with ethanol water：The ratio mixing of (10~20), extracts 2~4h, filtering retains filtrate, dense under the conditions of 80~90 DEG C
Contracting, is dried.
6. application according to claim 5, it is characterised in that its preparation method is：In terms of g/mL, by Japanese Raspberry Root herb
1 is pressed with 90% ethanol：15 ratio mixing, 3h is extracted under the conditions of 85 DEG C, is filtered, retains filtrate, is concentrated, is dried.
7. application according to claim 1, it is characterised in that the formulation of the fat-reducing medicament is oral formulations or injection system
Agent.
8. application according to claim 7, it is characterised in that the oral formulations are tablet, pill, granule, oral
Liquid or capsule.
9. application according to claim 7, it is characterised in that the ejection preparation is injection or injection powder injection.
10. application according to claim 1, it is characterised in that the fat-reducing medicament also includes other fat-reducing medicaments.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108096585A (en) * | 2018-02-28 | 2018-06-01 | 杨堃 | Application of the Japanese Raspberry Root extract as gene therapy sensitizer |
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| JP2006519231A (en) * | 2003-02-26 | 2006-08-24 | ユニゲン・ファーマシューティカルス・インコーポレーテッド | Formulations for the prevention and treatment of carbohydrate-induced diseases and conditions |
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Application publication date: 20171103 |