CN106798762A - One Plant Extracts and its preparation method and application - Google Patents
One Plant Extracts and its preparation method and application Download PDFInfo
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- CN106798762A CN106798762A CN201710170576.6A CN201710170576A CN106798762A CN 106798762 A CN106798762 A CN 106798762A CN 201710170576 A CN201710170576 A CN 201710170576A CN 106798762 A CN106798762 A CN 106798762A
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- Prior art keywords
- apiolin
- parts
- plant extracts
- glucosides
- glucose
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Classifications
-
- 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)
- A61K36/61—Myrtaceae (Myrtle family), e.g. teatree or eucalyptus
-
- 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)
-
- 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
-
- 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
-
- 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)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides Plant Extracts and its preparation method and application.The plant extracts is extracted by strophanthus divaricatus and Guava Leaf and obtained；In the plant extracts, contain the flavonoids active material for accounting for its weight more than 60%.The plant extracts can remove DPPH free radicals in vitro, can significantly reduce high lipid food and feed lipid of mice, can be effectively improved mice caused by dimethylbenzene xylene ear swelling inflammation, and isoprel can be protected to cause Effect On Rat Myocardial Injury.Described plant extracts come from it is among the people commonly use plant or medicinal and edible plant, it is safe and reliable, have no toxic side effect, can be used as new anti-oxidant, anti-inflammatory, the medicine or health food of lipid-loweringing.
Description
Technical field
The invention belongs to Chinese medicine, natural medicine field, more particularly, to Plant Extracts and preparation method thereof and
Using.
Background technology
Angiocardiopathy is to influence one of major disease of human health at present, and oxidative stress and inflammatory reaction are fallen ill at it
During occupy sizable ratio.
According to generally acknowledged damage theory, when low close in cholesterol, fat, cell waste and the blood plasma carried in blood
Degree lipoprotein (LDL) is piled up in areas of vessel damage：One side LDL is oxidized modification turns into ox-LDL；On the other hand, endothelium is thin
Born of the same parents' macrophage of seeking help is swallowed.Macrophages turn after phagocytosis turns into foam cells, and accumulation forms patch.Now, put down
Sliding myocyte is bred and is migrated covering patch formation fibrous cap, arterial channel is narrowed, the reduction of surrounding tissue oxygen content.
Foam cells is unstable, and necrose collaboration inflammation, produces spot grumeleuse, and surrounding tissue blood supply functional deterioration is dead, causes tissue
Necrosis, so as to trigger the diseases such as coronary heart diseases and angina pectoris, myocardial infarction and cerebrovas-cularaccident.
So eliminating excess activity oxygen radical, regulation dyslipidemia, suppress inflammatory reaction, to reducing atherosclerosis
Property angiocardiopathy generation, development play the role of it is important.But existing medicament categories with above-mentioned effect are limited, part
There is side effect, it is therefore desirable to further expand the optional species of medicine.
The content of the invention
It is an object of the invention to according to deficiency of the prior art, there is provided a Plant Extracts.The plant carries
Take in thing containing flavonoids active compound, damage, suppress inflammatory reaction, regulation fat with treating or alleviating active oxygen radical
Metabolic disorder, Ischemic myocardium equivalent damage really, are expected to as a kind of effective anti-oxidant, anti-inflammatory, reducing blood lipid and the protection heart
Blood vessel drug eluting or the health food with above-mentioned effect.
Preparation method another object of the present invention is to provide above-mentioned plant extracts.
Application another object of the present invention is to provide above-mentioned plant extracts.
Above-mentioned purpose of the present invention is achieved through the following technical solutions：
The invention provides a Plant Extracts, the plant extracts is strophanthus divaricatus and Guava Leaf after alcohol extracting
Obtain, containing the flavonoids active material at least accounting for its quality more than 60% in the plant extracts, the flavonoids activity
Material includes following component：
Apiolin, Vitexina, Saponaretin, apiolin -6,-C- glucosides of 8- bis-, apiolin -6-C- glucose -8-
C- rhamnosides, apiolin -6-C- rhamnose -8-C- glucosides, Isoschiaftoside, celery
Dish element -6-C- glucose sugar -8-C- Arab glycosides, apiolin -6-C- xylose -8-C- glucosides, apiolin -6-C- grapes
Sugar -8-C- xylosides, Kaempferide, Kaempferol -3-O- β-D glucosides, Isorhamnetin, Isorhamnetin -3-O- β-D rutinoses
Glycosides, Quercetin, Quercetin -3-O- β-D- arabopyranose glycosides, guaijaverin, Reynoutrin, Hyperoside, isoquercitrin.
Preferably, in the flavonoids active material, 0.2~1 part of apiolin, 3~10 parts of Vitexina, Saponaretin are contained
2~8 parts, apiolin -6,1~4 part of bis--C- glucosides of 8-, 0.5~2 part of apiolin -6-C- glucose -8-C- rhamnosides,
0.4~2 part of apiolin -6-C- rhamnose -8-C- glucosides, Isoschiaftoside 0.3~2
Part, 1.2~3.2 parts of glycosides of apiolin -6-C- glucose sugar -8-C- Arab, apiolin -6-C- xylose -8-C- glucosides 0.4
~2 parts, 0.3~2 part of apiolin -6-C- glucose -8-C- xylosides, 0.4~2 part of Kaempferide, Kaempferol -3-O- β-D grapes
1~3 part of glucosides, 0.3~1.2 part of Isorhamnetin, 6~12 parts of Isorhamnetin -3-O- β-D rutinosides, Quercetin 0.5~1.2
Part, 5~11 parts of Quercetin -3-O- β-D- arabopyranoses glycosides, 6~15 parts of guaijaverin, 3~10 parts of Reynoutrin, Hypericum Chinense
1.5~4.5 parts of glycosides, 1.5~4.5 parts of isoquercitrin.
Preferably, in the flavonoids active material, 0.52 part of apiolin, 8.2 parts of Vitexina, Saponaretin 5.5 are contained
Part, apiolin -6,2.2 parts of bis--C- glucosides of 8-, 0.91 part of apiolin -6-C- glucose -8-C- rhamnosides, apiolin -
0.97 part of 6-C- rhamnose -8-C- glucosides, 0.9 part of Isoschiaftoside, apiolin -6-
2.3 parts of glycosides of C- glucose sugar -8-C- Arab, 0.86 part of apiolin -6-C- xylose -8-C- glucosides, apiolin -6-C- Portugals
0.99 part of grape sugar -8-C- xylosides, 1.12 parts of Kaempferide, 2.3 parts of Kaempferol -3-O- β-D glucosides, Isorhamnetin 0.98
Part, 10.7 parts of Isorhamnetin -3-O- β-D rutinosides, 0.79 part of Quercetin, Quercetin -3-O- β-D- arabopyranose glycosides
8.8 parts, 10.3 parts of guaijaverin, 6.4 parts of Reynoutrin, 2.5 parts of Hyperoside, 2.1 parts of isoquercitrin.
The structure of the flavone compound in the flavonoids active material is as shown in table 1：
The structural formula of flavone compound in the plant extracts of the present invention of table 1:
Preferably, the extracting method of the plant extracts is specially:
It is 1: 5~5: 1 mixing according to mass ratio by strophanthus divaricatus and Guava Leaf, alcohol is carried out using 30%~80% ethanol
Carry, solid-liquid ratio is 1: 10~30, crude extract is concentrated under reduced pressure, centrifugation, supernatant adds 3~5 times of volume of water after with 0.5~1 times of medicinal material
The macroporous resin adsorption of weight is overnight；Eluted using alcoholic solution, collect 40~90% alcohol elution fractions, gel is crossed after being concentrated under reduced pressure
Post separation, the eluant, eluent that uses of separation be 60%~95% methanol aqueous solution, merging flavonoids stream part, it is concentrated under reduced pressure after with pure
Change, wash-out collects corresponding stream part, merges, and concentration, vacuum drying obtains the plant extracts.
Preferably, gel column is LH-20 sephadex columns, is purified with ODS column chromatographys after being concentrated under reduced pressure.
Most preferably, with 0.5kg strophanthus divaricatus and 0.5kg Guava Leafs as raw material, it is extracted, separate, lived after purification
Property extract position.Extraction solvent is 70% ethanol, and 1: 20,90 DEG C of solid-liquid ratio is heated to reflux；Crude extract is concentrated under reduced pressure into alcohol taste
After light, 3000r/min room temperatures centrifugation 20min, supernatant uses 1000g macroporous resin adsorptions overnight after adding 4 times of volume purified waters；After
And eluted with 10%, 70%, 100% methanol solution, 70% methanol-eluted fractions are collected, cross LH-20 glucans after being concentrated under reduced pressure
Gel post separation, eluant, eluent is 90% methanol aqueous solution, and polyam ide TLC chromatogram tracking merges flavonoids stream part, is concentrated under reduced pressure
Purified with ODS column chromatographys afterwards, aqueous methanol gradient wash-out collects corresponding stream part, merges, and concentration, 65 DEG C of vacuum drying are obtained
Plant extracts.
Application of the plant extracts in preventing and treating active oxygen radical damage medicine or health food is prepared.
Application of the plant extracts in the medicine or health food of preventing and treating hyperlipidemia is prepared.
Application of the plant extracts in the medicine or health food of preventing and treating atherosclerosis is prepared.
Application of the plant extracts in preventing and treating diabetes microvascular damage medicine or health food is prepared.
Compared with prior art, the present invention has advantages below and beneficial effect：
Containing the flavone compound no less than 60% mass percent in plant extracts provided by the present invention,
And empirical tests its can remove DPPH free radicals and active oxygen radical in vitro, experimental animal blood fat, Er Qieneng can be significantly reduced
Protection isoprel causes acute myocardial injury rat model, can be effectively improved lipopolysaccharides (LPS) induction inflammatory model mouse
Inflammatory reaction.Described plant extracts come from it is among the people commonly use plant or medicinal and edible plant, it is safe and reliable, it is nontoxic secondary to make
With can be used as a kind of novel oxidation-resistant, anti-inflammatory, the health food of lipid-loweringing or pharmaceutical preparation raw material.
Brief description of the drawings
Fig. 1 is plant extracts MpPgFF preparation technology general flow charts.
Fig. 2 is that plant extracts MpPgFF removes DPPH free radical results.
Fig. 3 is influences of the plant extracts MpPgFF to hyperlipemia model mouse liver tectology.
Fig. 4 is influences of the plant extracts MpPgFF to myocardial infarction and ischemia model rat heart muscle pathology tectology.
Specific embodiment
The present invention is further illustrated below in conjunction with specific embodiments and the drawings, but embodiment does not do any to the present invention
The restriction of form.Unless stated otherwise, the reagent for using of the invention, method and apparatus are the art conventional reagent, method
And equipment.
Unless stated otherwise, agents useful for same of the present invention and material are purchased in market.
Embodiment 1：
It is extracted, separate, lived after purification as shown in figure 1, with 0.5kg strophanthus divaricatus and 0.5kg Guava Leafs as raw material
Property extract position.Extraction solvent is 70% ethanol, and 1: 20,90 DEG C of solid-liquid ratio is heated to reflux；Crude extract is concentrated under reduced pressure into alcohol taste
After light, 3000r/min room temperatures centrifugation 20min, supernatant uses 1000g macroporous resin adsorptions overnight after adding 4 times of volume purified waters；After
And eluted with 10%, 70%, 100% methanol solution, 70% methanol-eluted fractions are collected, cross LH-20 glucans after being concentrated under reduced pressure
Gel post separation, eluant, eluent is 90% methanol aqueous solution, and polyam ide TLC chromatogram tracking merges flavonoids stream part, is concentrated under reduced pressure
Purified with ODS column chromatographys afterwards, aqueous methanol gradient wash-out, 254nm and 280nm wavelength detectings collect corresponding stream part, merge,
Concentration, 65 DEG C of vacuum drying obtain the 124.5mg of Objective extraction thing MpPgFF 1.
By determining, in above-mentioned Objective extraction thing, by mass percentage, containing flavonoids active material 69.34%, respectively
Flavone compound concrete content is：Apiolin (1,0.52%), Vitexina (2,8.2%), Saponaretin (3,5.5%), celery
- C- the glucosides of dish element -6,8- bis- (4,2.2%), apiolin -6-C- glucose -8-C- rhamnosides (5,0.91%), celery
Element -6-C- rhamnose -8-C- glucosides (6,0.97%), Isoschiaftoside (7,
0.9%), apiolin -6-C- glucose sugar -8-C- Arab's glycosides (8,2.3%), apiolin -6-C- xylose -8-C- glucosides
(9,0.86%), apiolin -6-C- glucose -8-C- xylosides (10,0.99%), Kaempferide (11,1.12%), Kaempferol -
3-O- β-D glucosides (12,2.3%), Isorhamnetin (13,0.98%), Isorhamnetin -3-O- β-D rutinosides (14,
10.7%), Quercetin (15,0.79%), Quercetin -3-O- β-D- arabopyranoses glycosides (16,8.8%), guaijaverin
(17,10.3%), Reynoutrin (18,6.4%), Hyperoside (19,2.5%), isoquercitrin (20,2.1%).
Embodiment 2：
It is extracted, separate, lived after purification as shown in figure 1, with 0.5kg strophanthus divaricatus and 1.0kg Guava Leafs as raw material
Property extract position.Extraction solvent is 50% ethanol, and 1: 20,90 DEG C of solid-liquid ratio is heated to reflux；Crude extract is concentrated under reduced pressure into alcohol taste
After light, depressurize suction filtration, and supernatant uses 1500g macroporous resin adsorptions overnight after adding 4 times of volume purified waters；Then with 10%, 70%,
100% methanol solution is eluted, and collects 70% methanol-eluted fractions, and LH-20 sephadex post separations, wash-out are crossed after being concentrated under reduced pressure
Agent is 90% methanol aqueous solution, and polyam ide TLC chromatogram tracking merges flavonoids stream part, pure with ODS column chromatographys after being concentrated under reduced pressure
Change, aqueous methanol gradient wash-out, 254nm and 280nm wavelength detectings collect corresponding stream part, merge, concentration, 65 DEG C of vacuum are done
It is dry to obtain the 630.8mg of Objective extraction thing MpPgFF 1.
By determining, in above-mentioned Objective extraction thing, by mass percentage, containing flavonoids active material 62.4%, respectively
Flavone compound concrete content is：Apiolin (1,0.22%), Vitexina (2,3.4%), Saponaretin (3,2.7%), celery
- C- the glucosides of dish element -6,8- bis- (4,1.16%), apiolin -6-C- glucose -8-C- rhamnosides (5,0.51%), celery
Dish element -6-C- rhamnose -8-C- glucosides (6,0.44%), Isoschiaftoside (7,
0.39%), apiolin -6-C- glucose sugar -8-C- Arab's glycosides (8,1.25%), apiolin -6-C- xylose -8-C- glucose
Glycosides (9,0.46%), apiolin -6-C- glucose -8-C- xylosides (10,0.37%), Kaempferide (11,0.43%), kaempferia galamga
Phenol -3-O- β-D glucosides (12,1.3%), Isorhamnetin (13,0.38%), Isorhamnetin -3-O- β-D rutinosides (14,
6.7%), Quercetin (15,1.19%), Quercetin -3-O- β-D- arabopyranoses glycosides (16,10.7%), guaijaverin
(17,14.6%), Reynoutrin (18,8.3%), Hyperoside (19,4.2%), isoquercitrin (20,3.7%).
Embodiment 3：
It is extracted, separate, lived after purification as shown in figure 1, with 1.0kg strophanthus divaricatus and 0.5kg Guava Leafs as raw material
Property extract position.Extraction solvent is 60% ethanol, and 1: 20,90 DEG C of solid-liquid ratio is heated to reflux；Crude extract is concentrated under reduced pressure into alcohol taste
After light, depressurize suction filtration, and supernatant uses 1500g macroporous resin adsorptions overnight after adding 4 times of volume purified waters；Then with 10%, 70%,
100% methanol solution is eluted, and collects 70% methanol-eluted fractions, and LH-20 sephadex post separations, wash-out are crossed after being concentrated under reduced pressure
Agent is 90% methanol aqueous solution, and polyam ide TLC chromatogram tracking merges flavonoids stream part, pure with ODS column chromatographys after being concentrated under reduced pressure
Change, aqueous methanol gradient wash-out, 280nm and 340nm wavelength detectings collect corresponding stream part, merge, concentration, 65 DEG C of vacuum are done
It is dry to obtain the 590.2mg of Objective extraction thing MpPgFF 1.
By determining, in above-mentioned Objective extraction thing, by mass percentage, containing flavonoids active material 65.7%, respectively
Flavone compound concrete content is：Apiolin (1,0.8%), Vitexina (2,9.2%), Saponaretin (3,6.0%), celery
- C- the glucosides of element -6,8- bis- (4,2.9%), apiolin -6-C- glucose -8-C- rhamnosides (5,1.2%), apiolin -
6-C- rhamnose -8-C- glucosides (6,1.5%), Isoschiaftoside (7,1.2%), celery
Dish element -6-C- glucose sugar -8-C- Arab's glycosides (8,3.1%), apiolin -6-C- xylose -8-C- glucosides (9,
1.8%), apiolin -6-C- glucose -8-C- xylosides (10,1.9%), Kaempferide (11,1.3%), Kaempferol -3-O- β-D
Glucoside (12,3.0%), Isorhamnetin (13,1.0%), Isorhamnetin -3-O- β-D rutinosides (14,11.7%), Mongolian oak
Pi Su (15,0.6%), Quercetin -3-O- β-D- arabopyranoses glycosides (16,5.3%), guaijaverin (17,6.2%) is auspicious
Promise glycosides (18,3.7%), Hyperoside (19,1.8%), isoquercitrin (20,1.5%).
Embodiment 4:Application experiment:
Extract MpPgFF removes DPPH free radicals
1st, the measure of test solution preparation and free radical scavenging
The plant extracts MpPgFF for obtaining is extracted in Example 1, is dissolved with DMSO, be configured to the mother of 14.5mg/mL
Liquid.100 μ L mother liquors accurately are pipetted, respectively with 50,100,120,150,200 times of methanol dilution, the MpPgFF of various concentrations is obtained final product
Need testing solution.
The MpPgFF test liquids of the 100 above-mentioned concentration gradients of μ L are taken respectively in brown tool plug bottle, then are added thereto to 3mL
DPPH standard liquids (precision weighs 2.5mg DPPH, with methanol constant volume in 100mL brown volumetric flasks), mix.It is dark in room temperature
Place stands 25min, is then designated as At in 517nm mensuration absorbance values, and experiment is in triplicate.DPPH free radicals are calculated according to the following formula
Clearance rate：E=(A0-At)/A0 × 100% (A0:The absorbance of DPPH standard liquids；At：After adding need testing solution t
Absorbance).Using IC50(when clearance rate is 50%, the concentration of test sample) is the evaluation index of radical scavenging activity size.
Extract MpPgFF is to the clear rate measurement result of DPPH free radicals as shown in Fig. 2 its DPPH free radical clear activity
IC50 about 0.038mg/mL, illustrate that it has preferable radical scavenging activity.The extract of embodiment 2 and embodiment 3
MpPgFF has the activity similar with the extract of embodiment 1.
Embodiment 5:Application experiment:
Extract MpPgFF improves mice caused by dimethylbenzene xylene auricle inflammatory swelling
1 experiment material：
Dimethylbenzene：Tianjin great Mao chemical reagent factories；Aspirin：Guangzhou Quan Ao chemical companies；
The plant extracts MpPgFF for obtaining is extracted in Example 1：High, medium and low dosage is respectively 200mg/kg,
100mg/kg, 50mg/kg.
2 experimental animals：
SPF grades of Kunming mouse, male and female half and half, 18~22g of weight is provided by Guangdong Medical Lab Animal Center,
Credit number：SCXK (Guangdong) 2008-0002.
3 experimental techniques：
50 mouse, are randomly divided into 5 groups：Model control group, positive controls (aspirin, 100mg/kg), extract
MpPgFF high doses group (200mg/kg), middle dose group (100mg/kg), low dose group (50mg/kg).Each group mouse stomach is given
Give the medicine of corresponding dosage, administered volume 20ml/kg.One time a day, continuous 7d, 1h after last dose, and 100 μ L dimethylbenzene are equal
Even to be applied to mouse right ear two sides, left ear puts to death the de- cervical vertebra of mouse after 1h as control, cuts two ears of left and right, with a diameter of
The auricle that 8mm card punch removes at Bilateral Symmetry claims quality, and the difference using left and right auricle quality calculates different doses as swelling
Measure the swelling inhibiting rate of administration group.
The extract MpPgFF paraxylene of table 2 causes the influence (n=10) of mice auricle swelling
Compare with model group,*P ﹤ 0.05,**P ﹤ 0.01
Test result indicate that, compared with control group, although extract MpPgFF low dose groups are not statistically significant, but have
Mitigate the trend of inflammatory swelling, its high, middle dose group can substantially mitigate mice caused by dimethylbenzene xylene auricle inflammatory swelling, and dosage
Inhibiting rate higher is bigger.The extract MpPgFF of embodiment 2 and embodiment 3 has the activity similar with the extract of embodiment 1.
Embodiment 6:Application experiment:
Extract MpPgFF reduction high fat diet guidance model lipid of mice
1 experiment material：
Cholesterol, NaTDC：Beijing Ding Guo Bioisystech Co., Ltd；Sucrose：Shantou City's brilliance laboratory；Tween
80：Tianjin great Mao chemical reagent factories；Propylthiouracil (PTU)：GuangDong HuaNan Pharmacy Group Co., Ltd；Blood fat recovery capsule：Beijing North
Great Wei Xin bio tech ltd；T-CHOL (TC), triglycerides (TG), HDL (HDL-C), low density lipoprotein
Albumen (LDL-C) detection kit：Zhongsheng Beikong Biological Science ＆ Technology Co., Ltd., lot number is respectively 121321,114961,
110431 and 110441.
The plant extracts MpPgFF for obtaining is extracted in Example 1：High, medium and low dosage is respectively 90mg/kg, 60mg/
Kg, 30mg/kg.
2 experimental animals：
SPF grades of Kunming mouse, male, 18~22g of weight is provided by Guangdong Medical Lab Animal Center, license
Card number：SCXK (Guangdong) 2008-0002.
3 experimental techniques：
SPF grades of body weight is the male mice 72 of 18~22g, is divided into 6 groups, including Normal group, model comparison
Group, positive controls (blood fat recovery capsule), MpPgFF high doses group, MpPgFF middle dose groups, MpPgFF low dose groups, mark respectively
Note.
Each group gives normal water and feed daily, and in addition to normal group, remaining each group every morning is filled with fat emulsion
Stomach (fat emulsion by cholesterol, lard, NaTDC, propylthiouracil (PTU), Tween-80, sucrose, distilled water is by a certain percentage
Prepare, melted with preceding 37 DEG C of water-baths, gavage volume 20ml/kg), hyperlipemia mice model is set up, afternoon is again respectively with difference
Liquid gavage (gavage volume 20ml/kg), is carried out continuously 14d.Claim within every 2 days 1 body weight, monitoring Mouse Weight change.Each group is moved
After thing last time gavage, water is can't help in fasting, and 12h posterior orbits take blood, and blood sample is centrifuged 10min under the conditions of 4 DEG C, 3000rpm, point
From serum, -20 DEG C save backup, and another day survey blood parameters.
Using lipid determination kit, by specification detects serum total cholesterol (TC), triglycerides (TG), high density fat
The content of albumen (HDL-C) and low-density lipoprotein (LDL-C).
Separately take liver to weigh, calculate mouse liver index (liver index=liver weight/body weight × 100)；Hepatic tissue is taken to be placed in
Soaked in 10% formaldehyde, FFPE, section, HE normal dyeings, each group hepatic pathology tissue shape is observed under an optical microscope
The change of state.
4 experimental results：
The extract MpPgFF of table 3 to hyperlipemia model mouse TC, TG, HDL and LDL influence (N=12)
Compare with normal group,*P ﹤ 0.05,**P ﹤ 0.01；Compare with model group,△P ﹤ 0.05,△△P ﹤ 0.01
The extract MpPgFF of table 4 to hyperlipemia in mice liver index influence (N=12)
Hepatic pathology morphologic observation：
The hepatic sinusoid of normal mouse liver is enriched, and comparatively dense occurs without fat accumulation phenomenon (Fig. 3 A) in cytoplasm.Model
There is mild fatty denaturation in group mouse liver cell, locally there is fat droplets packing phenomenon (Fig. 3 B).Gavage extract MpPgFF 2
Zhou Hou, hyperlipidemic mice liver fat quantity has reduction in various degree, with high dose group most pronounced effects (Fig. 3 D), low dosage effect
It is really poor, cell infiltration, hepatic cell cords disorder (Fig. 3 F).Effects of Xuezhikang group steatosis is lighter, but is dispersed in cell infiltration
(Fig. 3 C).The extract MpPgFF of embodiment 2 and embodiment 3 has the activity similar with the extract of embodiment 1.
Embodiment 7:Application experiment：
Extract MpPgFF protection isoprels cause acute myocardial injury
1st, experimental drug and reagent
Isoproterenol hydrochloride injection (ISO)：Shanghai Hefeng Pharmaceutical Co., Ltd. produces, lot number：110202；Hydration
Chloral：Solution on Chemical Reagents in Shanghai company of Chinese Medicine group, lot number：20050205；Superoxide dismutase (SOD), MDA
(MDA), glutathione peroxidase (GSH-PX), Coomassie brilliant blue protein determination kit：Biological reagent is built up in Nanjing to be had
Limit company produces；Compound danshen dripping pills：Tianjin Tasly Pharmaceutical Co., Ltd produces, lot number：110703；Extract
MpPgFF。
2nd, experimental animal
SPF grades of SD rat, male and female half and half, weight (200 ± 20) g is provided by Guangdong Medical Lab Animal Center, is moved
Thing credit number：SCXK (Guangdong) 2008-0002.
3rd, instrument
MedLab-6.0 System of organism signal；WFZ-26A ultraviolet-uisible spectrophotometers；MS-18 is full-automatic
Biochemical Analyzer；TGL-16G desk centrifuges；NIKON E600 microscopes.
4th, method：Animal packet and medication
SD rats 60, are randomly divided into 6 groups, respectively every group 10, Normal group (normal group), model control group
(model group), Composite Salvia Dropping Pill group (324mg/kg), MpPgFF high doses group (90mg/kg), middle dose group (60mg/kg),
Low dose group (30mg/kg).Each administration group is daily by body weight gastric infusion 1 time, continuous 5d, Normal group and model control group
Give the distilled water of same volume.
5th, isoprel causes Acute Myocardial Ischemia Rats model to prepare
1h after above-mentioned each group rat last dose, with 10% chloraldurate (0.3ml/100g) intraperitoneal injection of anesthesia, lies on the back
Position is fixed, connects MedLab-6.0 System of organism signal ECG electrodes, first records one section of normal ECG.With
Afterwards, in addition to normal group, each group rat takes the subcutaneous i.e. ISO of multi-point injection isoprel (2mg/kg), Normal group
The physiological saline of equivalent is injected, multi-point injection position includes：Injection is finished in the common 5-6 points in four limbs root and back, 10s, is recorded
1 after injection, 3,5,8,10,15,20,25, the ECG of 30min rats, count 1 respectively, 3,5,8,10,15,20,25,30min
ECG J points move down value, and (J points are the terminal portion minute and the ST sections of interface point of starting, the i.e. terminal of S ripples of QRS complex, with PR sections as base
Line, the changing value of J point displacements).After having surveyed electrocardiogram, in addition to normal group, every rat (is noted in abdominal cavity to 2 days ISO5mg/kg again
Penetrate), while continue gastric infusion, once a day.
6th, Testing index
After rat last gives ISO 24h, abdominal aortic blood, centrifugation prepares serum, using automatic clinical chemistry analyzer
Detection serum in LDH) and CK activity.Separately take the part cardiac muscle descended at 1/3 in rat heart and be placed in 10% formaldehyde and soak solid
It is fixed, serial section after FFPE, HE dyeing, light Microscopic observation pathological change；The same position cardiac muscular tissue of the rat apex of the heart separately is taken,
Use ice normal saline flushing, filter paper to be weighed after blotting in right amount, be fully ground under condition of ice bath and be made the homogenate of 10% tissue, survey
The content of SOD, MAD, GSH-PX.Tissue protein content is determined using Coomassie Brilliant Blue.
7th, statistical procedures
All data with mean ± standard deviation () represent, analyzed and processed using the statistical softwares of SPSS 13.0, compare between group
One-way analysis of variance is relatively used, with P<0.05 expression difference has conspicuousness.
8th, result
Influences of the extract MpPgFF to Acute Myocardial Ischemia Rats Pathomorphologic
From Fig. 4 results, rats in normal control group myocardial tissue structure is complete, and cardiac muscle fibre is in short cylinder, and cardiac muscle is thin
Born of the same parents' marshalling, fine and close, nucleus ovalize, centrally located, size is homogeneous, endochylema even dyeing (Fig. 4 A).Model group is big
There is popularity myofibrosis cordis, necrosis, telangiectasis, between cardiac muscle fibre in mouse cardiac muscle fibre arrangement disorder, cardiac muscle
Oedema and massive inflammatory cells infiltrated (Fig. 4 B).Some are disorderly for Composite Salvia Dropping Pill group rat heart muscle fiber alignment, indivedual cardiac muscles
There is cardiac myocyte hypertrophy, local myocardial fibre modification, necrosis (Fig. 4 C).MpPgFF high dose group rat heart muscles tissue cardiac muscle is fine
, there is individually cardiac muscular tissue's point sheet necrosis and denaturation (Fig. 4 F) in slight oedema between dimension, and local a small amount of inflammatory cell infiltration.
, partly there is cardiac muscular tissue's point sheet necrosis and denaturation, a small amount of inflammatory of interstitial in middle dose group major part cardiac muscle fibre marshalling
Cellular infiltration (Fig. 4 E).There is the necrosis of cardiac muscular tissue sheet and denaturation in low dose group, telangiectasis, between cardiac muscle fibre in
Degree oedema, the obvious inflammatory cell infiltration of interstitial (Fig. 4 D).
Influences of the MpPgFF to Acute Myocardial Ischemia Rats Serum fibrosis markers
Compared with normal group, CK, LDH content are significantly raised (P ＜ 0.01) in model group rats serum, represent heavy dose of
ISO can cause cardiac muscle cell's ischemic injuries.Compare with model group, composition is high, middle dose group Serum LDH, CK contents are obvious
Reduce (P ＜ 0.05).Prompting combination thing has improves myocardial cell injury, mitigates the effect of myocardial ischemia.Compound danshen dripping pills
Group Serum LDH, CK are significantly reduced (P ＜ 0.01).The results are shown in Table 5.
The MpPgFF of table 5 to rats with myocardial ischemia serum CK and LDH influence (N=10)
Compare with normal group：△P<0.05,△△P<0.01；Compare with model group：★P<0.05,★★P<0.01；
Influences of the MpPgFF to Acute Myocardial Ischemia Rats response to oxidative stress
The result of table 6 shows that compare with normal group, MDA contents are significantly raised in model group rats cardiac muscular tissue, SOD contents
It is remarkably decreased (P ＜ 0.01), GSH-PX vigor is remarkably decreased (P ＜ 0.05), represents that the rat model myocardial tissue oxidizing stress
Increased response, lipid peroxidation product increases.Compared with model group, MpPgFF is high, middle dose group MDA contents are substantially reduced, together
When SOD contents, GSH-PX vigor it is significantly raised.Prompting MpPgFF can significantly improve cardiac muscle confrontation lipid while resisting myocardial ischemia
Peroxidization ability.The extract MpPgFF of embodiment 2 and embodiment 3 has the activity similar with the extract of embodiment 1.
The MpPgFF of table 6 to rats with myocardial ischemia cardiac muscular tissue SOD, MDA and GSH-PX influence (N=10)
Compare with normal group：△△P<0.01；Compare with model group：★P<0.05,★★P<0.01。
Claims (9)
1. Plant Extracts, it is characterised in that the plant extracts is that strophanthus divaricatus and Guava Leaf are obtained after alcohol extracting
, the flavonoids active material at least accounting for its quality more than 60%, the flavonoids active material are contained in the plant extracts
Including following component：
Apiolin, Vitexina, Saponaretin, apiolin -6,-C- glucosides of 8- bis-, apiolin -6-C- glucose -8-C- mouse
Lee's glucosides, apiolin -6-C- rhamnose -8-C- glucosides, Isoschiaftoside, celery
Element -6-C- glucose sugar -8-C- Arab glycosides, apiolin -6-C- xylose -8-C- glucosides, apiolin -6-C- glucose -
8-C- xylosides, Kaempferide, Kaempferol -3-O- β-D glucosides, Isorhamnetin, Isorhamnetin -3-O- β-D rutinosides, Mongolian oak
Pi Su, Quercetin -3-O- β-D- arabopyranose glycosides, guaijaverin, Reynoutrin, Hyperoside, isoquercitrin.
2. plant extracts according to claim 1, it is characterised in that in the flavonoids active material, contain apiolin
0.2 ~ 1 part, 3 ~ 10 parts of Vitexina, 2 ~ 8 parts of Saponaretin, apiolin -6,1 ~ 4 part of bis--C- glucosides of 8-, apiolin -6-C-
0.5 ~ 2 part of glucose -8-C- rhamnosides, 0.4 ~ 2 part of apiolin -6-C- rhamnose -8-C- glucosides, apiolin -6-C-
0.3 ~ 2 part of arabinose -8-C- glucosides, 1.2 ~ 3.2 parts of glycosides of apiolin -6-C- glucose sugar -8-C- Arab, celery
0.4 ~ 2 part of element -6-C- xylose -8-C- glucosides, 0.3 ~ 2 part of apiolin -6-C- glucose -8-C- xylosides, Kaempferide 0.4
~ 2 parts, 1 ~ 3 part of Kaempferol -3-O- β-D glucosides, 0.3 ~ 1.2 part of Isorhamnetin, Isorhamnetin -3-O- β-D rutinosides 6 ~
12 parts, 0.5 ~ 1.2 part of Quercetin, 5 ~ 11 parts of Quercetin -3-O- β-D- arabopyranoses glycosides, 6 ~ 15 parts of guaijaverin, Rui Nuo
3 ~ 10 parts of glycosides, 1.5 ~ 4.5 parts of Hyperoside, 1.5 ~ 4.5 parts of isoquercitrin.
3. plant extracts according to claim 1, it is characterised in that in the flavonoids active material, contain apiolin
0.52 part, 8.2 parts of Vitexina, 5.5 parts of Saponaretin, apiolin -6,2.2 parts of bis--C- glucosides of 8-, apiolin -6-C- Portugals
0.91 part of grape sugar -8-C- rhamnosides, 0.97 part of apiolin -6-C- rhamnose -8-C- glucosides, apiolin -6-C- I
Primary 0.9 part of sugar -8-C- glucosides, 2.3 parts of glycosides of apiolin -6-C- glucose sugar -8-C- Arab, apiolin -6-C- xyloses -
0.86 part of 8-C- glucosides, 0.99 part of apiolin -6-C- glucose -8-C- xylosides, 1.12 parts of Kaempferide, Kaempferol -3-
2.3 parts of O- β-D glucosides, 0.98 part of Isorhamnetin, 10.7 parts of Isorhamnetin -3-O- β-D rutinosides, Quercetin 0.79
Part, 8.8 parts of Quercetin -3-O- β-D- arabopyranoses glycosides, 10.3 parts of guaijaverin, 6.4 parts of Reynoutrin, Hyperoside 2.5
Part, 2.1 parts of isoquercitrin.
4. the plant extracts according to any one of claims 1 to 3, it is characterised in that extracting method is specially：By strophanthus divaricatus
According to mass ratio it is 1: 5~5: 1 mixing with Guava Leaf, alcohol extracting is carried out using 30%~80 % ethanol, solid-liquid ratio is 1: 10~
30, crude extract is concentrated under reduced pressure, centrifugation, supernatant adds 3 ~ 5 times of volume of water after with 0.5~1 times of macroporous resin adsorption of medicinal material weight
Overnight；Eluted using alcoholic solution, collect 40~90% alcohol elution fractions, gel post separation is crossed after being concentrated under reduced pressure, separate what is used
Eluant, eluent is 60%~95% methanol aqueous solution, merges flavonoids stream part, and with purifying after being concentrated under reduced pressure, wash-out collects corresponding stream part,
Merge, concentration, vacuum drying obtains the plant extracts.
5. plant extracts according to claim 4, it is characterised in that gel column is LH-20 sephadex columns, is depressurized dense
Purified with ODS column chromatographys after contracting.
6. plant extracts described in any one of claims 1 to 3 is preparing preventing and treating active oxygen radical damage medicine or health care food
Application in product.
7. plant extracts described in any one of claims 1 to 3 is in the medicine or health food of preventing and treating hyperlipidemia is prepared
Using.
8. plant extracts described in any one of claims 1 to 3 prevents and treats the medicine or health food of atherosclerosis in preparation
In application.
9. plant extracts described in any one of claims 1 to 3 is preparing preventing and treating diabetes microvascular damage medicine or health care food
Application in product.
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Cited By (5)
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|---|---|---|---|---|
| CN107721993A (en) * | 2017-11-06 | 2018-02-23 | 长沙爱扬医药科技有限公司 | A kind of method that Vitexina and Saponaretin are extracted from strophanthus divaricatus |
| CN109908162A (en) * | 2017-12-12 | 2019-06-21 | 广州白云山和记黄埔中药有限公司 | A kind of pharmaceutical composition and its preparation and medical usage for treating depression |
| CN111620917A (en) * | 2020-05-08 | 2020-09-04 | 广西壮族自治区中国科学院广西植物研究所 | Isovitexin-2' -O-beta-D-glucopyranoside, and preparation method and application thereof |
| CN111759924A (en) * | 2020-08-16 | 2020-10-13 | 华明剑 | Traditional Chinese medicine composition with anti-aging effect |
| CN114558121A (en) * | 2022-03-11 | 2022-05-31 | 烟台新时代健康产业有限公司 | Composition for protecting respiratory tract and lung injury and preparation method and application thereof |
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2017
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| 孙冬梅等: "布渣叶化学成分和药理作用研究进展", 《世界中医药》 * |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107721993A (en) * | 2017-11-06 | 2018-02-23 | 长沙爱扬医药科技有限公司 | A kind of method that Vitexina and Saponaretin are extracted from strophanthus divaricatus |
| CN109908162A (en) * | 2017-12-12 | 2019-06-21 | 广州白云山和记黄埔中药有限公司 | A kind of pharmaceutical composition and its preparation and medical usage for treating depression |
| CN109908162B (en) * | 2017-12-12 | 2022-10-28 | 广州白云山和记黄埔中药有限公司 | Pharmaceutical composition for treating depression, preparation and medical application thereof |
| CN111620917A (en) * | 2020-05-08 | 2020-09-04 | 广西壮族自治区中国科学院广西植物研究所 | Isovitexin-2' -O-beta-D-glucopyranoside, and preparation method and application thereof |
| CN111759924A (en) * | 2020-08-16 | 2020-10-13 | 华明剑 | Traditional Chinese medicine composition with anti-aging effect |
| CN114558121A (en) * | 2022-03-11 | 2022-05-31 | 烟台新时代健康产业有限公司 | Composition for protecting respiratory tract and lung injury and preparation method and application thereof |
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