CN109078064A - A kind of Fructus Rubi extract and its preparation method and application - Google Patents
A kind of Fructus Rubi extract and its preparation method and application Download PDFInfo
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- CN109078064A CN109078064A CN201810990835.4A CN201810990835A CN109078064A CN 109078064 A CN109078064 A CN 109078064A CN 201810990835 A CN201810990835 A CN 201810990835A CN 109078064 A CN109078064 A CN 109078064A
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- fructus rubi
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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)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- 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
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Gastroenterology & Hepatology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Nutrition Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
It the present invention relates to a kind of Fructus Rubi extract and its preparation method and application, specifically discloses and uses raspberry medicinal material for raw material, the Fructus Rubi extract successively is prepared by ultrasonic alcohol extracting, concentration and freeze-drying；And specifically disclose application of the Fructus Rubi extract in the health care product or drug for preparing anti-liver organization oxidation, protection liver organization damage.Fructus Rubi extract of the present invention can significantly promote GSH content; enhancement of SOD activity; it is obviously improved CAT activity; reduce MDA content; it can be substantially reduced ALT, AST and LDH activity, i.e. Fructus Rubi extract has exploitation at the prospect for the health care product or drug for resisting liver oxidation and protection hepar damnification.
Description
Technical field
The invention belongs to pharmaceutical technology field, it is related to a kind of Fructus Rubi extract and its preparation method and application, specifically relates to
And a kind of Fructus Rubi extract and preparation method thereof and anti-liver aoxidize and protect hepar damnification in application.
Background technique
According to the World Health Organization in 2016 issue statistics show China every year there are about 400,000 people die of it is related with hepatitis
Complication.According to statistics, by the end of 2016, there are 90,000,000 Chronic Hepatitis Bs in China, wherein and 28,000,000 people need to treat,
7000000 people need emergency treatment because of major Liver disease and pathogenesis of cancer risk；At the same time, fatty liver and alcoholic liver at
For the second largest hepatopathy, total number of patients reaches 1.2 hundred million people, and wherein male occupies 80% or more ratio.Almost in every 13
Compatriots just have the liver of 1 people problematic, this is closely bound up with bad life habits, such as excessive drinking, poor eating habits, excessive labor
Tired and long-time is stayed up late, in addition, sedentary, blindness vegetarian diet and weight-reducing are all the reason of causing hepatopathy.
Liver is an organ in body based on metabolic function, and deoxidation, storage glycogen are played inside body
The effects of with the synthesis of secreted protein；Liver also manufactures dead bile in digestive system；Liver is in digestion
Maximum glandula digestive, adult human liver averagely weigh 1.5 kilograms；Liver is the major organs of urea synthesizing, is the weight of metabolism
Want organ.Liver is our important organs, and liver is bad, can seriously affect our health.Liver is the main solution of body
Malicious organ such as stays up late, indulges in excessive drinking, medication, infection etc., liver was in labourer when there is the case where aggravating body purification burden
Make state, function of detoxification is impaired over time, and cells of organs accelerates aging, and content greatly increases body endotoxin in blood,
Self deterioration also causes the aging of other organs simultaneously.Hepatic blood flow is reduced when liver aging, and liver cell quantity falls sharply, and liver becomes
To cirrhosis, weight is decreased obviously, and gradually appears the lesions such as liver fat, liver inflammation, hepatic cirrhosis, liver fibrosis and canceration
Situation.
Currently, the main means for liver protecting include: movement appropriate, fat extra in liver is excluded in time；
Body and mind loosens；Careful medication reduces liver toxin expelling burden；Keep good lifestyle habits；Take liver protection health food.
Raspberry is a kind of xylophyta of rose family rubus, has dual-purpose of drug and food double effects.Fresh fruit warp
Drug, as traditional Chinese medicine raspberry is made in processing, and " raspberry, smell sweet and neutral is nontoxic, beneficial for the record of Compendium of Material Medica document
Kidney astringent sperm contracting urine yang invigorating functions "；" explaining abstruseness of the canon of materia medica " record " raspberry, it is sweet to put down into kidney, play a sun and control impotence, controlling nocturnal emission with astringent drugs takes the photograph excessive, strong kidney and nothing
It is scorching it is partially, controlling nocturnal emission with astringent drugs and undoubtedly puckery evil, the product of gold and jade "；" raspberry is to nourish Kidney-Yin medicine, taste for " materia medica " record
With slightly sour, the negative gas for taking the photograph dissipation can be received and production of sperm liquid ".
Modern study confirms that raspberry has the function such as kidney-nourishing, the emerging impotence of establishing-Yang, brain tonic and intelligence development, beautifying face and moistering lotion and anti-curing cancers
Effect, but there is no the report for aoxidizing and protecting hepar damnification related efficacy for anti-liver in relation to raspberry.
Summary of the invention
The present invention compensates for the shortcomings of the prior art, provides a kind of Fructus Rubi extract and preparation method thereof and answers
With.
In order to achieve the above objectives, the present invention adopts the following technical scheme:
A kind of preparation method of Fructus Rubi extract is provided according to an aspect of the present invention, uses raspberry medicinal material for original
Alcohol solution is added in material, and ultrasonic alcohol extracting is prepared at 10-50kHz, 200-800W and 30-70 DEG C.
In the above-mentioned technical solutions, the preparation method specifically includes: taking the raspberry medicinal material of clean dried, pulverizes and sieves
Raspberry powder is obtained, the ethanol water that concentration of volume percent is 50-90% then is added by 10-30mL/g liquid-to-solid ratio, stirs
It mixes uniformly, ultrasonic alcohol extracting 15-60min, extracts 2 times, make after combined extract at 20-40kHz, 300-700W and 40-60 DEG C
It is standby to obtain.
Further, in the above-mentioned technical solutions, during ultrasonic alcohol extracting, the liquid-to-solid ratio is 15-24mL/g.
Preferably, in the above-mentioned technical solutions, during ultrasonic alcohol extracting, the percent by volume of the ethanol water is dense
Degree is 72-85%.
Preferably, in the above-mentioned technical solutions, during ultrasonic alcohol extracting, supersonic frequency, the ultrasound of the ultrasonic extraction
Power, Extracting temperature and extraction time are respectively 20-25kHz, 450-550W, 42-48 DEG C and 35-42min.
Still further, in the above-mentioned technical solutions, the particle size of the raspberry powder is 600-1000 mesh.
Preferably, in the above-mentioned technical solutions, the preparation method further includes after ultrasonic alcohol extracting to by alcohol extracting mixture
Filtering, concentration, lyophilization and grinding, the cryogenic temperature and cooling time of the lyophilization be respectively -30~-60 DEG C and
30-45min。
The Fructus Rubi extract that preparation method is prepared is provided according to a further aspect of the invention, it is specifically, described to cover
The partial size of basin extract is 150-300 mesh, wherein the content of raspberry flavones is 0.72-1.86wt%.
Fructus Rubi extract is provided according to a further aspect of the invention is preparing anti-liver organization oxidation, protection liver organization
The health care product of damage or the application in drug.
Specifically, the liver organization oxidation or damage are aoxidized or are damaged for the liver organization caused by concanavalin A.
Further, the liver organization oxidation includes the reduction of GSH content, SOD reduced activity, CAT activity reduce and MDA
The oxidation of liver organization caused by content increases, the liver organization damage include that ALT activity, AST activity and LDH activity raising are drawn
The liver organization damage risen.
Health care product or drug of the present invention, using Fructus Rubi extract as single active ingredient or as active constituent it
One, when its as active constituent for the moment, it is optional and other aoxidize effective in anti-liver organization, protect liver organization damage
Active constituent, such as kudzu root extract collectively form the active constituent of health care product or drug, to obtain ideal synergistic effect.
In addition to the active ingredient (s, health care product or drug of the present invention further include pharmaceutically acceptable at least one load
Body.The carrier understands by those skilled in the art, including but not limited to filler, disintegrating agent, lubricant, solubilising
Agent, suspending agent, wetting agent, solvent, emulsifier, stabilizer, antioxidant or surfactant etc., those skilled in the art can be according to
It is selected according to the actual demand of preparation.
Health care product or drug of the present invention can be prepared into oral preparation or ejection preparation via conventional method.Administration
Optional oral, the sublingual, vein of mode, subcutaneous, transdermal or local administration etc., give animal or people with unit dosage fonn.
In health care product or composite medicine of the invention, Fructus Rubi extract is usually prepared with dosage unit.Every dosage
Unit contains 4.8 to 12.6 grams of Fructus Rubi extracts, gives daily 1 time or multiple.Under specific condition can also using higher or
Relatively low-dose, the suitable dose of each patient is by doctor according to the age of mode of administration and the patient, weight and reaction come most
It determines eventually.
Fructus Rubi extract provided by the present invention has the advantage that
(1) the present invention provides the new applications of Fructus Rubi extract, can significantly promote GSH-Px activity, and enhancement of SOD is living
Property, it is obviously improved CAT activity, MDA content is reduced and is brought in addition, ALT, AST and LDH activity can be substantially reduced for hepatopath
Gospel；
(2) Fructus Rubi extract provided by the present invention is used to resist liver oxidation and the effect of protection hepar damnification is aobvious
It writes, has no toxic side effect；
(3) preparation process of Fructus Rubi extract provided by the present invention and for anti-liver organization oxidation, protection liver
The health care product of tissue damage or the preparation process of drug are simple, and technique is controllable, are easily converted to clinical application, have good answer
Use prospect.
Detailed description of the invention
Fig. 1 is influence result of the Fructus Rubi extract of the embodiment of the present invention to Gsh In Mouse Liver peptide (GSH) content
Comparative diagram；
Fig. 2 is that Fructus Rubi extract of the embodiment of the present invention is active to total number born in mouse liver (SOD)
Influence result comparative diagram；
Fig. 3 is that Fructus Rubi extract of the embodiment of the present invention ties the active influence of catalase in mouse liver (CAT)
Fruit comparative diagram；
Fig. 4 is influence result pair of the Fructus Rubi extract of the embodiment of the present invention to malonaldehyde in mouse liver (MDA) product
According to figure；
Fig. 5 is that Fructus Rubi extract of the embodiment of the present invention ties the active influence of glutamic-pyruvic transaminase in mice serum (ALT)
Fruit comparative diagram；
Fig. 6 is that Fructus Rubi extract of the embodiment of the present invention ties the active influence of glutamic-oxalacetic transaminease in mice serum (AST)
Fruit comparative diagram；
Fig. 7 is that Fructus Rubi extract of the embodiment of the present invention ties the active influence of lactic dehydrogenase in mice serum (LDH)
Fruit comparative diagram；
Note: compared with blank group, # represents p < 0.05, and ## represents p < 0.01；
Compared with model group, * represents p < 0.05, and * * represents p < 0.01.
Specific embodiment
With reference to the accompanying drawings and examples, specific embodiments of the present invention will be described in further detail.Implement below
Example is merely to illustrate the present invention, the protection scope being not intended to limit the invention.
Raspberry medicinal material used is commercial product in the embodiment of the present invention.
Embodiment 1
A kind of preparation method of Fructus Rubi extract, comprising the following steps:
S1, raspberry medicinal material is taken, removal of impurities washing is dried, and is crushed and is crossed 800 meshes, obtain raspberry powder；
S2, the ethanol water that concentration of volume percent is 70% is added by 20mL/g liquid-to-solid ratio in raspberry powder, stirred
It mixes uniformly, the ultrasonic extraction 45min at 25kHz, 450W and 45 DEG C is extracted 2 times；
S3, the alcohol extracting mixture in step S2 is filtered, obtains raspberry alcohol extract, being then concentrated to get volume is raspberry
The raspberry concentrate of the 1/8-1/6 of alcohol extract volume；
S4, raspberry concentrate is freeze-dried 25min at -30~-40 DEG C, after pulverizing cross 200 meshes to get
Fructus Rubi extract.
Through detecting, raspberry ketone content is 1.25wt% in Fructus Rubi extract obtained in above-described embodiment 1.
Embodiment 2
A kind of preparation method of Fructus Rubi extract, comprising the following steps:
S1, raspberry medicinal material is taken, removal of impurities washing is dried, and is crushed and is crossed 800 meshes, obtain raspberry powder；
S2, the ethanol water that concentration of volume percent is 70% is added by 25mL/g liquid-to-solid ratio in raspberry powder, stirred
It mixes uniformly, the ultrasonic extraction 30min at 35kHz, 500W and 50 DEG C is extracted 2 times；
S3, the alcohol extracting mixture in step S2 is filtered, obtains raspberry alcohol extract, being then concentrated to get volume is raspberry
The raspberry concentrate of the 1/8-1/6 of alcohol extract volume；
S4, raspberry concentrate is freeze-dried 35min at -40~-50 DEG C, after pulverizing cross 200 meshes to get
Fructus Rubi extract.
Through detecting, raspberry ketone content is 1.18wt% in Fructus Rubi extract obtained in above-described embodiment 2.
Embodiment 3
A kind of preparation method of Fructus Rubi extract, comprising the following steps:
S1, raspberry medicinal material is taken, removal of impurities washing is dried, and is crushed and is crossed 800 meshes, obtain raspberry powder；
S2, the ethanol water that concentration of volume percent is 70% is added by 30mL/g liquid-to-solid ratio in raspberry powder, stirred
It mixes uniformly, the ultrasonic extraction 50min at 25kHz, 550W and 55 DEG C is extracted 2 times；
S3, the alcohol extracting mixture in step S2 is filtered, obtains raspberry alcohol extract, being then concentrated to get volume is raspberry
The raspberry concentrate of the 1/8-1/6 of alcohol extract volume；
S4, raspberry concentrate is freeze-dried 40min at -50~-60 DEG C, after pulverizing cross 200 meshes to get
Fructus Rubi extract.
Through detecting, raspberry ketone content is 1.82wt% in Fructus Rubi extract obtained in above-described embodiment 3.
In order to verify the effect for resisting liver oxidation and protection hepar damnification of above-mentioned Fructus Rubi extract, above-mentioned reality is used
It applies the Fructus Rubi extract that the method provided in example 1 is prepared and has carried out pharmacodynamic experiment, detailed process is as follows:
1, experimental animal:
Healthy male BALB/c mouse 84, weight 17-19g, SPF grade are purchased from Chinese Academy of Sciences experimental animal institute, lot number:
SYXK (capital) -0020.
2, experiment reagent and instrument:
Superoxide dismutase SOD kit, cat catalase kit, glutathione GSH kit, malonaldehyde
MDA kit and ELISA kit are purchased from Nanjing and build up biological Co., Ltd.
3, experimental group and experimental method
The mouse bought adaptive feeding in gnotobasis, 20.5-21.5 DEG C of environment temperature, humidity 40-60%, by 12
Hour illumination, 12 hours no light alternatings, ad lib and drinking-water.Stochastic averagina is divided into 6 groups after a week for raising, and every group 14,
Respectively blank group (Blank), Con A model group (Con A, canavaline), positive controls (BC, bicyclic alcohols 200mg/
Kg), Fructus Rubi extract low dose group (FPZ-L, 200mg/kg), Fructus Rubi extract middle dose group (FPZ-M, 400mg/kg)
With Fructus Rubi extract high dose group (FPZ-H, 800mg/kg), shown in table 1 specific as follows.
Drug and dosage contrast table used in 1 each group mouse of table
As shown in table 1, positive controls (BC, 200mg/kg), Fructus Rubi extract low dose group (FPZ-L, 200mg/
Kg), Fructus Rubi extract middle dose group (FPZ-M, 400mg/kg) and Fructus Rubi extract high dose group (FPZ-H, 800mg/
Kg it) by design dosage stomach-filling, is administered 7 days, after the last administration, is deprived of food but not water altogether, ConA solution is subcutaneously injected using tail portion
200mg/kg is sampled after 8h to each group mouse modeling (in addition to blank group).
3.1 changes of weight
After the continuous 1 week stomach-filling drug of each group mouse, every diurnal variation of weight is as shown in table 2 below.
Every diurnal variation contrast table of 2 each group mouse weight of table
Note: (1) numerical value is indicated with the average value ± SD of 14 mouse
(2) * * p < 0.01, * p < 0.05, compared with model group
From the results shown in Table 2, compared with blank group, slow growth trend is presented in Con A group mouse weight, covers basin
The intervention of son is consistent with positive drug bicyclic alcohols, does not improve weight caused by Con A and increases slow effect, this and protect liver, reduction
Fat accumulation effect is consistent；The body weight increase trend of raspberry middle dose group is consistent with positive control bicyclic alcohols trend, raspberry
High dose group has the function of stronger control mouse weight rapid growth.
3.2 internal organ coefficients
Con A modeling carries out anaesthetic treatment to each group mouse after 8h, and tail portion takes dissection removal liver and spleen after blood, raw
After reason salt water is got express developed, clean filter paper suck dry moisture, rapid weighing, the results are shown in Table 3.
3 each group mice viscera index contrast table of table
Note: (1) numerical value is indicated with the average value ± SD of 14 mouse
(2) * * p < 0.01, * p < 0.05, withModel groupIt compares
From the results shown in Table 3, Con A will cause liver inflammation, cause liver enlargement, can be seen that from data, lead
Cause liver enlargement 30%, Fructus Rubi extract can very well liver-protective form it is normal, in addition, 3 result of comparative analysis table can
To find out, the effect of Fructus Rubi extract is more prominent compared with bicyclic alcohols, and liver can be kept to be in normal condition, may be derived from and cover
Basin extract is natural materials, has more safety compared with combination drug.
3.3 anti-liver organization oxidabilities
The mouse liver tissue of taking-up is ground using buffer, utilizes superoxide dismutase SOD, hydrogen peroxide
The kit of enzyme CAT, glutathione GSH and malonaldehyde MDA carry out the measurement of index of correlation.
Total protein is organized to determine
It prepares standard protein concentration series: taking standard protein BSA (3.2mg/mL), it is continuous etc. using physiological saline as solvent
Than the standard protein series for being diluted to 1.6,0.8,0.4,0.2,0.1mg/mL；
The preparation of BCA working solution: it is calculated according to sample size, adds 1 volume BCA reagent B (50:1) by 50 volume BCA reagent As
Appropriate BCA working solution is prepared, is mixed well；
Protein sample control is standby: 5 μ L protein sample liquid taken, 20 μ L normal saline dilutions are added, it is to be measured；
Sample system prepares: according to detection architecture design sequentially add on 96 orifice plates various concentration titer, sample liquid,
Each 5 μ L of distilled water, 25 μ L BCA working solutions mix well；Each sample sets 6 in parallel.
Protein content detection: sample ELISA Plate is placed in 37 DEG C of incubators and is incubated for 30min, is quickly put into microplate reader,
The sample well of distilled water is added as reference, in 562nm wavelength detecting OD value.
3.3.1, the influence of GSH content
It is as shown in Figure 1 the influence result comparative diagram of each group Gsh In Mouse Liver peptide (GSH) content.
The result of comparison diagram 1, which can be seen that Con A, has the function of destroying liver function have the data of model group can be with
Confirm the success of modeling；ConA has the function of that GSH is promoted in inducing mouse body, and the protection that this is embodied in itself of mouse is special
Property, and Fructus Rubi extract GSH intracorporal to ConA model mice has the influence of highly significant, can significantly promote GSH
Activity；In addition, Fructus Rubi extract to the protection of liver oxidative damage more preferably compared with bicyclic alcohols, raspberry low dose group and the positive are right
It is suitable according to group (bicyclic alcohols) effect, and raspberry middle dose group and raspberry high dose group are compared with the effect of positive controls (bicyclic alcohols)
Power promotes 20-40%.
3.3.2, the active influence of T-SOD
It is illustrated in figure 2 the active influence result comparative diagram of total number born (SOD) in each group mouse liver.
The result of comparison diagram 2 can be seen that Fructus Rubi extract have to the liver SOD activity of Con A model group mouse it is aobvious
The castering action of work, specifically, the function and effect and positive controls of raspberry middle dose group and raspberry high dose group are (bicyclic
Alcohol) quite, and the function and effect of raspberry low dose group are weaker.
SOD is antioxidase important in organism, is the primary substance that free radical is removed in organism, in oxidative stress
In the aging of initiation, the activity of SOD is significantly affected, a large number of studies show that, in aged animal model and inflammation in animals mould
In the research such as type, being decreased obviously for liver organization SOD content is had been found that, illustrate that SOD level with liver aging is positively related.?
In this experimental example, the SOD enzyme activity in Con A model mice liver is significantly lower than control group, illustrates that aging model is set up, covers basin
After seed extract is intervened, SOD activity is enhanced in liver organization, illustrates that the endogenous antioxygen of rat liver can be improved in raspberry
Change ability.
3.3.3, the active influence of CAT
It is illustrated in figure 3 the active influence result comparative diagram of catalase (CAT) in each group mouse liver.
The result of comparison diagram 3 can be seen that Fructus Rubi extract have to the liver CAT activity of Con A model group mouse it is aobvious
The castering action of work.CAT is the key index of aging research, and increasing or decreasing for CAT can reflect indirectly body cell by freedom
The severity of base attack, CAT research is more in aging research, and the downstream effects enzyme as Antioxidation Mechanism, often with
SOD occurs simultaneously, to illustrate delaying senility function mechanism.Which kind of confirmed in research on anti-senescence, either aging model, mould
Endogenous anti-oxidative enzyme SOD and the CAT activity of formula biology can be used as the reliable finger of aging degree all as aging state changes
Mark.In this experimental example, after Fructus Rubi extract acts on Con A model group mouse, CAT activity all has increasing in liver organization
By force.CAT is a kind of detoxication enzyme, is present in the peroxide body of cell, and the content in liver cell is significantly larger than other groups
It knits, so it is this reason that the experimental results, which show as liver's activities of antioxidant enzymes, which to be changed greatly, also side light is for this
Any liver can have efficient degradation detoxication.
3.3.4, the influence of MDA product
It is illustrated in figure 4 the influence result comparative diagram of malonaldehyde (MDA) product in each group mouse liver.
The result of comparison diagram 4, which can be seen that Fructus Rubi extract, has the function of preventing Peroxidation Product in liver from generating,
The result of Fig. 4 is made a concrete analysis of it is found that Con A can lead to MDA product in mouse liver largely generates, the intervention of Fructus Rubi extract
MDA generation can be reduced.
MDA is one of important products of lipid peroxidation, is the sensitive indicator of measurement machine free radical metabolism, objective anti-
The level of interior free yl is reflected, reducing MDA content is an important mechanisms in anti-oxidant research.In a large amount of peroxidating model
In it can be observed that in serum and histocyte MDA a large amount of generations and deposition, MDA deposition excessively, gradually causes cellular damage
And organ damage, stiff reaction is formed, the organ damages such as liver are caused.In this experimental example, Con A causes in mouse liver tissue
MDA content increases, and illustrates that Con A causes the destruction of mouse body Antioxidant Enzyme Systems, antioxidant ability of organism weakens, and oxidation is answered
It increases sharply strong, excessive free radicals induce lipid peroxidation injury, and then have promoted the cell even damage and aging of organ, raspberry
Extract can obviously lower MDA product in liver, illustrate that Fructus Rubi extract can be by anti-oxidative damage, and then protect
The health of liver organ.
3.4 protection hepar damnification abilities
3.4.1 glutamic-pyruvic transaminase (ALT) active influence
The whole blood sample of mouse is placed 60 minutes at room temperature and enables its Spontaneous Condensation, is then centrifuged under 4 DEG C, 3000rpm
10 minutes, take top serum as analysis of material, using ELISA kit measurement glutamic-pyruvic transaminase (ALT) activity.As a result as schemed
Shown in 5, all numerical value are indicated with the average value ± SD of 14 mouse.
The result of comparison diagram 5 can be seen that glutamic-pyruvic transaminase (ALT) activity of normal mouse liver generally in 20-30U/
L, after Con A modeling, mouse liver ALT is steeply risen, and be can reach 7000U/L (Con A model group), is shown canavaline pair
Mouse liver causes apparent injury, significant inflammatory reaction occurs；Fructus Rubi extract influences obviously ALT, can be obvious
Reduce ALT activity.Compared with model group, raspberry low dose group, raspberry middle dose group and raspberry high dose group all have
The difference (p < 0.01) of highly significant, mouse can have very much under high dose Fructus Rubi extract (800mg/kg FPZ) intervention
Protect to effect liver from damage, glutamic-pyruvic transaminase (ALT) activity maintains identical level with positive drug group substantially.
3.4.2 glutamic-oxalacetic transaminease (AST) active influence
The whole blood sample of mouse is placed 60 minutes at room temperature and enables its Spontaneous Condensation, is then centrifuged under 4 DEG C, 3000rpm
10 minutes, take top serum as analysis of material, using ELISA kit measurement glutamic-oxalacetic transaminease (AST) activity.As a result as schemed
Shown in 6, all numerical value are indicated with the average value ± SD of 14 mouse.
The result of comparison diagram 6 can be seen that Fructus Rubi extract to glutamic-oxalacetic transaminease (AST) the activity tool reduced in serum
Play the role of clearly crossing, compared with Con A model group, the intervention of variant dosage Fructus Rubi extract can significantly drop
The vigor (p < 0.01) of low AST, the function and effect of raspberry high dose group are compared with raspberry low dose group and raspberry middle dose group
It is more obvious and suitable with positive controls (bicyclic alcohols, 200mg/kg) function and effect.
Serum glutamic pyruvic transminase (ALT) and glutamic-oxalacetic transaminease (AST) activity are the characteristics indexes of liver health.ALT is
A kind of normal enzyme in liver, the amount for occurring under normal conditions in blood be it is fewer, liver cell have destruction or
It is to be easy for increasing for transaminase occur in the case where damaging；AST is primarily present in liver cell mitochondria, when liver occurs sternly
When weight necrosis or destruction, it can just cause the significant raising of glutamic-oxalacetic transaminease.
3.4.3 lactic dehydrogenase (LDH) active influence
The whole blood sample of mouse is placed 60 minutes at room temperature and enables its Spontaneous Condensation, is then centrifuged under 4 DEG C, 3000rpm
10 minutes, take top serum as analysis of material, using ELISA kit measurement lactic dehydrogenase (LDH) activity.As a result as schemed
Shown in 7, all numerical value are indicated with the average value ± SD of 14 mouse.
Lactic dehydrogenase (LDH) is one of the important enzyme system of sugared anerobic glycolysis and gluconeogenesis, can be catalyzed pyruvic acid and L- cream
Reduction and oxidation reaction between acid are the important indicators of liver diseases, and LDH is often shown in oxyhepatitis or chronic active hepatitis
It writes or moderate increases, susceptibility is slightly below ALT.LDH activity is significantly raised when liver cancer, and especially metastatic hepatic carcinoma increases more aobvious
It writes.
The result of comparison diagram 7, which can be seen that Fructus Rubi extract, there is protection to make the hepar damnification induced by Con A
With, and the effect of raspberry low dose group, raspberry middle dose group and raspberry high dose group all has conspicuousness (p < 0.01),
Wherein, raspberry high dose group (800mg/kg) effect is especially prominent, with positive controls (bicyclic alcohols, 200mg/kg) effect phase
When the function and effect of each dosage group do not differ significantly.
To sum up experimental study shows that Fructus Rubi extract is effective against liver oxidation, protects hepar damnification, can be significantly
GSH-Px activity is promoted, enhancement of SOD activity is obviously improved CAT activity, reduces MDA content, can be substantially reduced ALT, AST and LDH
Activity, and its preparation process is simple, technique is controllable, is easily converted to clinical application, has a good application prospect.
Finally, being not intended to limit the scope of the present invention the above is only preferred embodiment of the invention.It is all this
Within the spirit and principle of invention, any modification, equivalent replacement, improvement and so on should be included in protection model of the invention
Within enclosing.
Claims (10)
1. a kind of preparation method of Fructus Rubi extract, which is characterized in that use raspberry medicinal material for raw material, it is molten that alcohols is added
Liquid, ultrasonic alcohol extracting is prepared at 10-50kHz, 200-800W and 30-70 DEG C.
2. preparation method according to claim 1, which is characterized in that specifically include: the raspberry medicinal material of clean dried is taken,
It pulverizes and sieves to obtain raspberry powder, the ethanol water that concentration of volume percent is 50-90% then is added by 10-30mL/g liquid-to-solid ratio
Solution stirs evenly, and ultrasonic alcohol extracting 15-60min, is extracted 2 times at 20-40kHz, 300-700W and 40-60 DEG C, merges and extracts
It is prepared after liquid.
3. preparation method according to claim 2, which is characterized in that during ultrasonic alcohol extracting,
The liquid-to-solid ratio is 15-24mL/g；
And/or the concentration of volume percent of the ethanol water is 72-85%；
And/or supersonic frequency, ultrasonic power, Extracting temperature and the extraction time of the ultrasonic extraction be respectively 20-25kHz,
450-550W, 42-48 DEG C and 35-42min.
4. preparation method according to claim 2 or 3, which is characterized in that
The particle size of the raspberry powder is 600-1000 mesh；
And/or further include to by the filtering of alcohol extracting mixture, concentration, lyophilization and grinding after ultrasonic alcohol extracting, the freeze-drying is dry
Dry cryogenic temperature and cooling time is respectively -30~-60 DEG C and 30-45min.
5. the Fructus Rubi extract that the described in any item preparation methods of claim 1-4 are prepared, it is preferable that the raspberry
The partial size of extract is 150-300 mesh, wherein the content of raspberry flavones is 0.72-1.86wt%.
6. Fructus Rubi extract described in claim 5 is in the health care for preparing anti-liver organization oxidation, protection liver organization damage
Application in product or drug.
7. application according to claim 6, which is characterized in that the liver organization oxidation or damage is by concanavalin As
The liver organization oxidation or damage caused.
8. application according to claim 7, which is characterized in that the liver organization oxidation includes the reduction of GSH content, SOD
Liver organization oxidation caused by reduced activity, CAT activity reduce and MDA content increases, the liver organization damage include ALT living
Property, AST activity and LDH activity increase caused by liver organization damage.
9. application according to claim 6, which is characterized in that the health care product or drug are single with Fructus Rubi extract
Active constituent or as one of active constituent, it is preferable that the health care product or drug further include pharmaceutically acceptable at least one
Kind carrier.
10. according to the described in any item applications of claim 6-9, which is characterized in that the health care product or drug are oral preparation
Or ejection preparation.
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| CN114209744A (en) * | 2022-01-11 | 2022-03-22 | 安徽农业大学 | Cream for treating mastitis of dairy cows and preparation method thereof |
| CN116036161A (en) * | 2022-12-23 | 2023-05-02 | 温州医科大学 | Raspberry extract and preparation method and application thereof |
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Application publication date: 20181225 |