CN107184658A - Glossy privet flower extract, extracting method and its application in terms of prevention and treatment metabolic syndrome medicine is prepared - Google Patents
Glossy privet flower extract, extracting method and its application in terms of prevention and treatment metabolic syndrome medicine is prepared Download PDFInfo
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- CN107184658A CN107184658A CN201710274380.1A CN201710274380A CN107184658A CN 107184658 A CN107184658 A CN 107184658A CN 201710274380 A CN201710274380 A CN 201710274380A CN 107184658 A CN107184658 A CN 107184658A
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- glossy privet
- ethanol
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- extract
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- 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
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Abstract
The invention belongs to medicine and/or field of health care products, and in particular to a kind of glossy privet flower extract, extracting method and its application in terms of prevention and treatment metabolic syndrome medicine is prepared.Glossy privet flower extract of the present invention is that mixture more than one or both of the total medicinal extract of Ligustrum japonicum Thunb.flower obtained, 40% ethanol position and 60% ethanol position is extracted from Ligustrum japonicum Thunb.flower.The invention also discloses the extracting method of above-mentioned glossy privet flower extract.The present invention is found by experiment that：Glossy privet flower extract has the effect of prevention and treatment metabolic syndrome, the absorption of fat cell sugar can especially be promoted, suppress the effect that PECTORAL LIMB SKELETON breaks up and promotes fat cell lipid metaboli, diabetes medicament and prevention and improvement obesity drug are prevented and treated available for preparing.
Description
Technical field
The invention belongs to medicine and/or health product technology field, and in particular to a kind of glossy privet flower extract, extracting method and
Its prepare prevention and treatment metabolic syndrome medicine in terms of application.
Background technology
Glossy privet（Ligustrum lucidumAit.）For Oleaceae glossy privet platymiscium.Its fruit medicine is the fruit of glossy privet
（Ligustri Lucidi Fructus）, sweet, bitter, cold.Return liver and kidney channel.With nourishing liver and kidney, effect of improving eyesight black hair.For
The deficiency of liver-yin and kidney-yin, dizziness and tinnitus, soreness and weakness of waist and knees, poliosis, mesh secretly fails to understand, Heat Diabetes, osteopyrexia and fever.The current fruit of glossy privet is ground
Study carefully very many, chemical composition mainly has triterpenes, flavonoids, iridoids and phenylethanol etc..Pharmacological activity mainly has anti-
Oxidation, anti-inflammatory, liver protection, anticancer, hypoglycemic and immunological regulation etc..In addition, having been reported that the separation of glossy privet bark identifies 19 chemical combination
Thing, in addition to the compound structure type often having in glossy privet, is also obtained the chemical combination such as umbelliferone, benzoic acid, niacinamide
Thing.The research of Ligustrum japonicum Thunb.flower is relatively fewer.The composition of glossy privet flowers volatile oil is mainly alcohol, aldehyde, ester etc..Ligustrum japonicum Thunb.flower chemical composition is main
There are flavonoids and sterols.It is another that there are some researches show Ligustrum japonicum Thunb.flower general flavone has stronger removing DPPH free radicals and nitrite
Ability.At present and have no research on Ligustrum japonicum Thunb.flower in terms of metabolism syndrome.
Obesity is that body fat caused by many factors such as heredity, environment accumulates the excessive, chronic metabolic of increased weight
Property disease.It is closely related with diabetes B, dyslipidemia.The energy that the energy of intake exceedes consumption can cause the accumulation of fat.
The increase of adipose tissue is caused with volume increase by increasing for adipocyte number.The propagation of adipose tissue, differentiation are not normal
The accumulation of adipose tissue can be caused.Body fat tissue, which is excessively accumulated, can secrete a series of hormone and Adipocyte Factor interference
Insulin signal transduction in the cell, triggers insulin resistance, and insulin resistance refers to internal peripheral tissues（Muscle, fat）It is right
The sensitiveness reduction of insulin so that intake of the peripheral tissues to glucose is reduced, causes blood glucose rise, and then induce 2 types sugar
Urine disease.Also, insulin resistance runs through the whole process of diabetes B.The primary structure of adipose tissue is sweet in fat drips, fat drips
The content of oily three esters accounts for 95%.Triglycerides is decomposed into free fatty and glycerine in the presence of lipase, and lipolytic is most main
The effect wanted is energy supply.Fat drips are referred to as " energy converter " again, are increased to fat related subcellular structure, intracellular fat drips
Fat cell volume can be caused to increase greatly.Fat Basic of Biology is：The increase of intracellular fat drips cause cell volume increase and
Newborn adipocyte number increases.Therefore, the formation of research fat drips and the metabolism of triglycerides are likely to become solution problem of obesity
One of means.Fat cell originates from mesodermal stem cell, and its differentiation and development goes through several stages：Versatile stem cell,
Mesenchymal precursor, PECTORAL LIMB SKELETON, mature fat cell.3T3-L1 PECTORAL LIMB SKELETONs are thin from mouse embryo fibroblast
Born of the same parents, turn into pre-adipose cell lines through clonal expansion, energy directed differentiation is mature fat cell, is fully shown in the spy of body cell
Point, including morphologic change, the expression of a variety of lipid metabolism enzymes, extensive lipid accumulation etc., it is in vitro study fat cell
Ideal model.The incidence of disease of obesity and diabetes B constantly rises, it has also become global health problem.Exploitation is prevented and treated
The natural products meaning for treating obesity and diabetes B is particularly far-reaching.
The content of the invention
Present invention aims to overcome that prior art defect is pre- there is provided glossy privet flower extract, extracting method and its preparing
Application in terms of anti-and treatment metabolic syndrome medicine.
To achieve the above object, the present invention is adopted the following technical scheme that：
A kind of glossy privet flower extract, the glossy privet flower extract is the total medicinal extract of Ligustrum japonicum Thunb.flower, 40% ethanol position and 60% ethanol position
One or both of more than mixture.
The extracting method of above-mentioned glossy privet flower extract, be specially：Dry Ligustrum japonicum Thunb.flower is taken, with petroleum ether heating and refluxing extraction 1-
3 times, each refluxing extraction 1-3h, refluxing extraction terminates rear separation of solid and liquid, residue obtained to volatilize petroleum ether, with 70 ± 5% ethanol rooms
Temperature lower Soakage extraction 2-4 times, each Soakage extraction 2-4 days merges extract solution and concentrated, obtains the total medicinal extract of Ligustrum japonicum Thunb.flower；Ligustrum japonicum Thunb.flower
After total medicinal extract ethanol dissolves, separated with D101 types macroporous resin adsorption, successively with water, 20% ethanol, 40% ethanol, 60% ethanol,
95% ethanol gradient elution, after eluent concentration, respectively obtains water position, 20% ethanol position, 40% ethanol position, 60% ethanol portion
Position, 95% ethanol position.Glossy privet flower extract is exactly any in the total medicinal extract of Ligustrum japonicum Thunb.flower, 40% ethanol position and 60% ethanol position
One or more kinds of mixtures.
Application of the above-mentioned glossy privet flower extract in terms of prevention and treatment metabolic syndrome medicine is prepared.
Above-mentioned application, can be specifically：Glossy privet flower extract is in terms of prevention and treatment diabetes medicament is prepared
Using the especially application in terms of the medicine and/or health products that promote fat cell sugar absorption, can be further to prepare
Promote the application in terms of the medicine and/or health products that fat cell sugar absorbs under insulin-resistant states.
Above-mentioned application, specifically can also be：Glossy privet flower extract is in terms of preparing prevention and improving obesity drug
Application, i.e., with potential effect of weight reducing, especially prepare suppress PECTORAL LIMB SKELETON differentiation medicine and/or health products side
The application in face or be prepare promote fat cell lipid metaboli medicine and/or health products in terms of application.
Compound preparation can also be made with this area customary adjuvant compounding in above-mentioned glossy privet flower extract.The compound preparation
Formulation can be for tablet, granule, capsule, pill or injection etc..
Compared to the prior art, the present invention has been found by experiment that glossy privet flower extract by suppressing PECTORAL LIMB SKELETON point
Change, promote fat cell lipid metaboli, and promote fat cell sugar to absorb and improve insulin resistance, be likely to become 2 types for the treatment of
Diabetes, a kind of fat new compound, i.e., it is with potential effect that is hypoglycemic, improving insulin resistance.Therefore, may be used
It is used as preparing prevention and treatment metabolic syndrome medicine and/or health products, especially prevents and improve diabetes
Medicine, obesity drug.
Brief description of the drawings
Fig. 1 is the oil red differential stain result of control group；
Fig. 2 is the oil red differential stain result of the total medicinal extract of Ligustrum japonicum Thunb.flower；
Fig. 3 is the oil red differential stain result at the ethanol position of Ligustrum japonicum Thunb.flower 40%；
Fig. 4 is the oil red differential stain result at the ethanol position of Ligustrum japonicum Thunb.flower 60%；
Fig. 5 is the influence that glossy privet flower extract is acted on 3T3-L1 fat cells lipid metaboli；
Fig. 6 is glossy privet flower extract to the sugared absorption of 3T3-L1 fat cells；
Fig. 7 is the formation of induced by dexamethasone 3T3-L1 mature fat cell insulin resistances；
Fig. 8 is effect of the glossy privet flower extract to 3T3-L1 fat cells sugar absorption under insulin-resistant states.
Embodiment
Technical scheme is further discussed in detail with reference to embodiments, but protection scope of the present invention
It is not limited thereto.
In following embodiments, unless otherwise specified, the percentage of ethanol refers both to percent by volume.
Embodiment 1
A kind of glossy privet flower extract, the glossy privet flower extract is the total medicinal extract of Ligustrum japonicum Thunb.flower, 40% ethanol position medicinal extract and 60% ethanol
Mixture more than one or both of position medicinal extract.
The extracting method of above-mentioned glossy privet flower extract, specifically includes following steps：
1）475 g are taken to dry Ligustrum japonicum Thunb.flower, addition petroleum ether is to Ligustrum japonicum Thunb.flower is submerged, then heating and refluxing extraction 2 times, backflow every time is carried
Take after 2h, refluxing extraction terminate, filtering is residue obtained to volatilize petroleum ether, with Soakage extraction 3 times at room temperature of 70% ethanol（70% second
Alcohol addition is advisable with submerging residue）, each Soakage extraction 3 days, merge extract solution simultaneously concentrate, obtain the total medicinal extract of Ligustrum japonicum Thunb.flower；
2）The total medicinal extract of Ligustrum japonicum Thunb.flower with 50% ethanol dissolve after, with D101 types macroporous resin adsorption separate, successively with water, 20% ethanol,
40% ethanol, 60% ethanol, 95% ethanol gradient elution, each five column volumes of gradient elution, each mL of column volume 2000, elution
After liquid concentration, water position, 20% ethanol position 7g, 40% ethanol position 60g, 60% ethanol position 24g, 95% ethanol portion are respectively obtained
Position 16g.
The influence of application test 1, glossy privet flower extract to 3T3-L1 PECTORAL LIMB SKELETONs activity
Using 3T3-L1 PECTORAL LIMB SKELETONs as cell model, Ligustrum japonicum Thunb.flower prepared by above-described embodiment 1 is added in induction atomization
Extract, oil red O stain identification 3T3-L1 PECTORAL LIMB SKELETON differentiation degrees.
Instrument material：Instrument：Preparative high performance liquid chromatography instrument（Waters 2535Q）；Rotary Evaporators（EYELA, east
Capital physics and chemistry apparatus Co., Ltd.）；LC-3000 efficient liquid phases prepare column chromatography（Beijing Chuangxin Tongheng Science and Technology Co., Ltd.）；Am-
400 NMR spectrometer with superconducting magnet（Bruker）；Electronic balance（Mettler-Toledo companies of the U.S.）；TGL-16gR high speed desktops are cold
Freeze centrifuge（Anting Scientific Instrument Factory, Shanghai）；Double one side clean work station（Model：SW-CJ-2FD types producer：Suzhou is net
Change equipment Co., Ltd）；CO2gas incubator（Model：3111 types, producer：Thermo scientific）；Inverted microscope
（Ckx31 types, olympus）；Full temperature shaken cultivation case（The grand experimental facilities Co., Ltd of Nereid in HZP types）；Centrifugation device
（800 type Shanghai Surgical Operation Equipment Factory）；All size liquid-transfering gun（transferpette）；Digital camera（canon EOS450D）
Inverted microscope（Nikon ECLiPSE TS100）；The silica gel thin-layer plates of GF 254（Yantai Hui You silica gel development corporation, Ltd.）；40
~ 80 mesh silica gel, 200 ~ 300 mesh silica gel and silica gel H（Yantai Hui You silica gel development corporation, Ltd.）；Sephadex LH-20（Sweden
Pharmacia companies）；D101 type macroreticular resins（Tianjin sea light Chemical Co., Ltd.）；C-18（German Merk companies）；Remaining examination
Agent is that analysis is pure；Glucose estimation kit（ShangHai RongSheng Biology Pharmacy Co., Ltd, 20161105147）；Glucose mark
Quasi- product（ShangHai RongSheng Biology Pharmacy Co., Ltd, lot number：20161001）；Mouse free fatty acid determination reagent kit（Nanjing is gloomy
Bei Jia bio tech ltd, lot number：201609）；Hyclone（Zhejiang Tian Hang biotech inc, lot number：
20160923）；The high sugared nutrient solutions of DMEM（Beijing Suo Laibao scientific ＆ technical corporation, lot number：F08HV090）；Penicillin streptomycin mixed liquor
（Solarbio, lot number：20161029）；Pancreatin cell dissociation buffer（The green skies, numbering：C0210）；Dimethyl sulfoxide (DMSO)（sigma
SHBC3313V）；Insulin（sigma ）；Tetrazolium bromide（Shanghai Chinese blue Science and Technology Ltd.）；Oil red O（sigma
SLBP5248V）； IBMX（Sigma, BCBH1214V）；DeX（sigma BCBM4557V）.Rosiglitazone tablets（The permanent auspicious system in Chengdu
Medicine Co., Ltd）；Lovastatin capsule（Yangzijiang Pharmaceutical Group Co., Ltd, 20160109）；Cell line：3T3-L1 Development of Mouse Embryos
Tire fibroblast is purchased from Chinese Academy of Sciences's cell bank.It is public that Ligustrum japonicum Thunb.flower is collected in Guiyang City, Guizhou Province Huaxi District wetland in June, 2015
Garden, by Guizhou University, professor Zhang Qianjun is accredited as glossy privet（Ligustrum lucidumAit.）Flower.Sample is existing in Henan
Institute of Chinese materia medica of university.
Experimental method：
Cell culture is with freezing：
3T3-L1 PECTORAL LIMB SKELETONs, with the high sugared nutrient solutions of the DMEM containing 15% hyclone（Containing penicillin：10U/mL, streptomysin 10
μg/mL）In 37 DEG C, 5% CO2Cultivated in the CO2gas incubator of saturated humidity, 2 d change liquid once, when cell fusion to 80%
Afterwards, nutrient solution in blake bottle is abandoned, 0.25% trypsin solution 1mL, effect 1min or so is added, 2 times of volume nutrient solutions is added and terminates
Digestion, is blown and beaten attached cell into nutrient solution with piping and druming pipe.Cell suspension is moved into sterile centrifugation tube, 1000 rpm centrifugations 5
Min, abandons supernatant.Passage：The cell precipitation that 1 blake bottle is collected is dispelled with the new nutrient solutions of 1mL, by 1：2 ratios are passed on.Carefully
Born of the same parents freeze：The cell precipitation that 2 blake bottles are collected is with 1.2 mL frozen stock solutions（Hyclone containing 10%DMSO）Dispel to be transferred to and freeze
Pipe is put into program temperature reduction box, is then placed in -80 DEG C of refrigerator overnights, takes out cryopreservation tube and is put into preservation in liquid nitrogen container.
Cytotoxicity assay：
The 3T3-L1 PECTORAL LIMB SKELETONs of exponential phase are collected, concentration of cell suspension are adjusted, with every hole 2 × 105Density inoculation
In 96 well culture plates, 100 μ L cell suspensions are added per hole.Cell is placed in 37 DEG C, 5% CO2And the carbon dioxide of saturated humidity
24 h are cultivated in incubator, cell fusion are treated to 60%-70%, the medicine of concentration gradient is added, each plate sets Normal group, molten
Matchmaker's control group（0.1% DMSO）, extract administration group, if 6 multiple holes, continue cultivate 48 h after, per hole add 10 μ L MTT it is molten
Liquid（5 g/L）, continue to cultivate 4 h.Nutrient solution in hole carefully is sucked, 100 μ L DMSO are added per hole and put concussion and cultivate case（100
r/min）10 min of middle concussion, make crystal fully dissolve.The absorbance in each hole is measured at the nm wavelength of ELIASA 570（OD
Value）.
Table 1：The total medicinal extract of Ligustrum japonicum Thunb.flower, 60% ethanol position influence on 3T3-L1 PECTORAL LIMB SKELETONs cytoactive（n=6）
As shown in Table 1, Normal group is compared with vehicle control group absorbance, and difference is not statistically significant, and illustrates solvent
Influence is not produced on cell viability.The total medicinal extract of Ligustrum japonicum Thunb.flower, 60% ethanol position dosage group compared with Normal group absorbance,
Difference is not statistically significant, and illustrates the total medicinal extract of Ligustrum japonicum Thunb.flower, 60% ethanol position dosage group without cytotoxic activity, i.e. glossy privet
The total medicinal extract of Hua, 60% ethanol position can be set to dosage below 300 μ g/mL dosage ranges.
Table 2:Influence of the ethanol position of Ligustrum japonicum Thunb.flower 40% to 3T3-L1 PECTORAL LIMB SKELETONs activity（n=6）
As shown in Table 2, Normal group is compared with vehicle control group absorbance, and difference is not statistically significant, and illustrates solvent
Influence is not produced on cell viability.40% ethanol position dosage group is compared with Normal group absorbance, and difference does not have statistics
Meaning, illustrates the 40% all dosage groups in ethanol position without cytotoxic activity.That is 40% ethanol position is in 300 μ g/mL dosage models
Enclosing following can be set to dosage.
The influence that application test 2, glossy privet flower extract break up to 3T3-L1 PECTORAL LIMB SKELETONs
Take the logarithm the 3T3-L1 PECTORAL LIMB SKELETON Secondary Cultures in growth period, by 1.5 × 105Density be inoculated into 96 orifice plates, carefully
After born of the same parents' fusion completely, then the h of contact inhibition 48 changes A containing differentiating inducer（0.5 mM IBMX 、1 μM DeX、10 μg/
mL Insulin）The high sugared nutrient solutions of DMEM（Containing 15% hyclone）, while dosing（It is divided into blank group, model group, Lip river and cuts down him
Spit of fland group（50 μM）, administration group）.Change B containing differentiation-inducing agents after 3 d into（Contain 10 μ g/mL Insulin）The high sugar trainings of DMEM
Nutrient solution（Containing 15% hyclone）, cellar culture liquid is changed after 2 d, liquid is hereafter changed every other day, is cultivated to 12 d, is carefully absorbed nutrient solution,
Ice-cold PBS is washed 2 times, is dried, and the paraformaldehydes of 50 μ L 4% are added per hole and fix 40 min, PBS is washed 2 times, dried again, oil red is added
O dyes 30 min, and PBS is washed 2 times, and ultrapure to wash 2 times, digital camera is taken pictures under inverted microscope（100×）.100 are added per hole
After μ L isopropanols, 40 min absorbance is determined using ELIASA under 495 nm wavelength.As a result it is as follows.
Data processing：As a result represented using mean value ± SD values, data statistics is using SPSS19.0 software single factor tests variance point
Analysis method（One-Way ANOVA）Compare its significant difference.
Table 3：The influence that the total medicinal extract of Ligustrum japonicum Thunb.flower breaks up to 3T3-L1 PECTORAL LIMB SKELETONs（n=6）
From table 3, Fig. 1,2, compared with model group, when the total medicinal extract 300 of Lovastatin group, Ligustrum japonicum Thunb.flower, 100 μ g/mL, oil red
Dye level is significantly reduced（P＜ 0.05）, when Ligustrum japonicum Thunb.flower 33,11 μ g/mL, oil red dye level is significantly reduced（P＜ 0.01）.
Lovastatin breaks up inhibiting rate：35.27%, the total medicinal extract of Ligustrum japonicum Thunb.flower breaks up inhibiting rate and is respectively：18.39%、21.80%、
32.68%、31.62%.Illustrate that Lovastatin and the total medicinal extract of Ligustrum japonicum Thunb.flower can suppress the differentiation of PECTORAL LIMB SKELETON, illustrate that it has
Potential quality for preventing and improving obesity.
Table 4:Influence of the ethanol position of Ligustrum japonicum Thunb.flower 40% to cell differentiation before 3T3-L1（n=6）
From table 4, Fig. 1,3, all dosage group oil red dye levels in the ethanol position of Ligustrum japonicum Thunb.flower 40% do not change, and illustrate glossy privet
40% ethanol position is spent to suppress the differentiation of 3T3-L1 PECTORAL LIMB SKELETONs.
Table 5：The influence that the ethanol position of Ligustrum japonicum Thunb.flower 60% breaks up to 3T3-L1 PECTORAL LIMB SKELETONs（n=6）
From table 5, Fig. 1,4, the μ g/mL dosage group oil red dye levels of 60% ethanol position of Ligustrum japonicum Thunb.flower 300 are significantly reduced（P<
0.001）, illustrate that the μ g/mL dosage groups of 60% ethanol position of Ligustrum japonicum Thunb.flower 300 can suppress the differentiation of 3T3-L1 PECTORAL LIMB SKELETONs, wherein,
The μ g/mL dosage groups differentiation inhibiting rate of 60% ethanol position of Ligustrum japonicum Thunb.flower 300 is 24.24%.The ethanol position 100 of Ligustrum japonicum Thunb.flower 60%, 33 and
11 μ g/mL dosage group oil red dye levels do not change, and illustrate the ethanol position 100,33 of Ligustrum japonicum Thunb.flower 60% and 11 μ g/mL dosage
Group can not suppress the differentiation of 3T3-L1 PECTORAL LIMB SKELETONs.
The glossy privet flower extract of application test 3 is to 3T3-L1 fat cell lipid metaboli function influences
Take the logarithm the 3T3-L1 PECTORAL LIMB SKELETON Secondary Cultures in growth period, by 1.5 × 105Density be inoculated into 96 orifice plates, carefully
After born of the same parents' fusion completely, then the h of contact inhibition 48 changes A containing differentiating inducer（0.5 mM IBMX 、1 μM DeX、10 μg/
mL Insulin）The high sugared nutrient solutions of DMEM（Containing 15% hyclone）, differentiation-inducing agents B is changed into after 3 d（10 μg/mL
Insulin）The high sugared nutrient solutions of DMEM（Containing 15% hyclone）, the high sugared nutrient solutions of the DMEM containing 15% hyclone are changed after 2 d,
Hereafter liquid is changed every other day, is cultivated to 12 d, is induced to differentiate into mature fat cell.Mature fat cell packet, dosing（It is divided into blank
Group, Rosiglitazone group（9.52 μM）, administration group）, continue cultivate 48 h, sterile tube collect cells and supernatant, 2600 g from
The min of the heart 20, carefully collects supernatant.The content of free fatty is determined using mouse free fatty acid determination reagent kit.As a result such as
Under.
Fig. 5 is glossy privet flower extract to 3T3-L1 fat cell lipid metaboli function influences.As shown in Figure 5,100 μ g/mL female
The loyal total medicinal extract of Hua（TE）, the ethanol positions of 33 μ g/mL 40%（40%EF）Dosage group free fatty acid content is higher than blank group, difference
With statistical significance（P<0.05）, the ethanol position dosage group free fatty acid content of 11 μ g/mL Ligustrum japonicum Thunb.flowers 40% is higher than blank
Group, difference has statistical significance（P<0.001）, the ethanol positions of 11 μ g/mL 60%（60%EF）Dosage group free fatty acid content
Higher than blank group, difference has statistical significance（P<0.05）, illustrate the 100 total medicinal extract of μ g/mL Ligustrum japonicum Thunb.flowers, 33,11 μ g/mL 40%
Ethanol position, the ethanol positions of 11 μ g/mL 60% can promote triglycerides catabolism in fat cell to be free fatty, tool
Play the role of reduction fat cell in content of triglyceride, illustrate its can be used for promote fat cell lipid metaboli, with for
Prevention and the potential quality for improving obesity.
The influence that the glossy privet flower extract of application test 4 absorbs to 3T3-L1 mature fat cells sugar
PECTORAL LIMB SKELETON is induced to differentiate into mature fat cell, and experimentation is with application test 3.It is grouped dosing（Be divided into blank group,
Rosiglitazone group（9.52 μM）, administration group）, continue to cultivate 48 h, take nutrient solution supernatant to be detected by Glucose estimation kit and train
Glucose content in nutrient solution.
Fig. 6 is the total medicinal extract of Ligustrum japonicum Thunb.flower, 40% ethanol position, 60% ethanol position to the sugared absorption of 3T3-L1 fat cells.
It will be appreciated from fig. 6 that Rosiglitazone, 300, the 11 total medicinal extract of μ g/mL Ligustrum japonicum Thunb.flowers, the ethanol positions of 33,11 μ g/mL 40%, 100 μ g/mL
60% ethanol position glucose content is less than blank group, and difference has statistical significance（P<0.05）.The 33 total medicinal extract of μ g/mL, 33,
The ethanol position glucose contents of 11 μ g/mL 60% are less than blank group, and difference has statistical significance（P<0.01）, illustrate that Roger arranges
The total medicinal extract of the μ g/mL of ketone, 300,33 and 11,33 and the ethanol positions of 11 μ g/mL 40%, the ethanol positions of 100,33,11 μ g/mL 60%
Intake of the fat cell to periphery glucose can be promoted, cause glucose content reduction degree in culture medium to be more than blank group Portugal
Grape sugared content reduces degree, illustrates that it has the potential quality of prevention and treatment diabetes.
Application test 5, under insulin-resistant states glossy privet flower extract to 3T3-L1 mature fat cells sugar absorb
Influence
PECTORAL LIMB SKELETON is induced to differentiate into mature fat cell, and experimentation is with application test 3.Mature fat cell is divided into blank
Group and reactance group.Reactance group gives 1 μM of d of effects of dexamethasone 3, takes supernatant to determine glucose content, identifies pancreas islet
Element resistance model modeling success.Insulin resistance component group dosing（It is divided into blank group, Rosiglitazone group（20 μM）, administration group,
Supernatant is taken to determine sugared value.
Fig. 7 is the formation of induced by dexamethasone 3T3-L1 mature fat cell insulin resistances.As shown in Figure 7, resistance group
Glucose content is higher than model group, and difference has statistical significance（P<0.001）.Illustrate resistance group fat cell to periphery grape
The intake effect reduction of sugar, insulin resistant model modeling success.
Fig. 8 is the total medicinal extract of Ligustrum japonicum Thunb.flower, 40% ethanol position, 60% ethanol position to 3T3-L1 fat under insulin-resistant states
The effect that cell sugar absorbs.As shown in Figure 8, Rosiglitazone, 33, the 11 total medicinal extract of μ g/mL Ligustrum japonicum Thunb.flowers, the ethanol of 33,11 μ g/mL 60%
Position glucose content is less than blank group, and difference has statistical significance（P<0.05）.300th, the total medicinal extract glucose of 100 μ g/mL
Content is less than blank group, and difference has statistical significance（P<0.01）.40% ethanol position is compared with blank group, and difference is not united
Meter learns meaning.Illustrate, 33, the 11 total medicinal extract of μ g/mL Ligustrum japonicum Thunb.flowers, the 60% all dosage groups in ethanol position can promote insulin resistance
Intake of the fat cell to periphery glucose under state, illustrates to can be used for preventing and treating diabetes.
Conclusion：Glossy privet flower extract can suppress PECTORAL LIMB SKELETON differentiation, promote fat cell lipid metaboli, and promote fat
Fat cell sugar absorbs and improved insulin resistance, it is thus possible to a kind of novel compounds as treatment diabetes B, obesity
Thing, i.e., it is with potential effect that is hypoglycemic, improving insulin resistance.Therefore, it is used as preparing prevention and treatment
Metabolic syndrome medicine and/or health products, especially prevent and improve diabetes medicament, obesity drug.
Claims (8)
1. a kind of glossy privet flower extract, it is characterised in that the glossy privet flower extract is the total medicinal extract of Ligustrum japonicum Thunb.flower, 40% ethanol position
With mixture more than one or both of 60% ethanol position.
2. the extracting method of glossy privet flower extract described in claim 1, it is characterised in that take dry Ligustrum japonicum Thunb.flower, with petroleum ether plus
Circumfluence distillation 1-3 times, each refluxing extraction 1-3h, refluxing extraction terminates rear separation of solid and liquid, residue obtained to volatilize petroleum ether, use
70 ± 5% ethanol Soakage extraction 2-4 times at room temperature, each Soakage extraction 2-4 days merges extract solution and concentrated, obtains Ligustrum japonicum Thunb.flower total
Medicinal extract；After the total medicinal extract ethanol of Ligustrum japonicum Thunb.flower dissolves, separated with D101 types macroporous resin adsorption, successively with water, 20% ethanol, 40% second
Alcohol, 60% ethanol, 95% ethanol gradient elution, after eluent concentration, respectively obtain water position, 20% ethanol position, 40% ethanol portion
Position, 60% ethanol position, 95% ethanol position.
3. application of the glossy privet flower extract described in claim 1 in terms of prevention and treatment metabolic syndrome medicine is prepared.
4. application as claimed in claim 3, it is characterised in that the glossy privet flower extract is preparing prevention and treatment diabetes
Application in terms of medicine.
5. application as claimed in claim 4, it is characterised in that the glossy privet flower extract promotes fat cell sugar to inhale in preparation
Application in terms of the medicine and/or health products of receipts.
6. application as claimed in claim 3, it is characterised in that the glossy privet flower extract is preparing prevention and improving obesity
Application in terms of medicine.
7. application as claimed in claim 6, it is characterised in that the glossy privet flower extract is preparing suppression PECTORAL LIMB SKELETON point
Application in terms of the medicine and/or health products of change.
8. application as claimed in claim 6, it is characterised in that the glossy privet flower extract is preparing promotion fat cell fat generation
Application in terms of the medicine and/or health products thanked.
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| CN201710274380.1A CN107184658B (en) | 2017-04-25 | 2017-04-25 | Ligustrum lucidum extract, extraction method and application thereof in preparation of medicines for preventing and treating metabolic syndrome |
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| CN107184658B CN107184658B (en) | 2020-10-27 |
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