CN106474145A - Application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera - Google Patents
Application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera Download PDFInfo
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- CN106474145A CN106474145A CN201611071660.4A CN201611071660A CN106474145A CN 106474145 A CN106474145 A CN 106474145A CN 201611071660 A CN201611071660 A CN 201611071660A CN 106474145 A CN106474145 A CN 106474145A
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- China
- Prior art keywords
- polysaccharides
- leaves
- moringa
- moringa oleifera
- food
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- C—CHEMISTRY; METALLURGY
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- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
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Abstract
The invention belongs to field of natural product chemistry, disclose a kind of application in preparation preventing and treating alcoholic liver injury medicine and food for Polysaccharides from Leaves of Moringa oleifera.This Polysaccharides from Leaves of Moringa oleifera is slightly carried using decoction and alcohol sedimentation technique, carries out purification in conjunction with technological processes such as 14000rpm centrifugation, deproteinization, AB 8 type macroporous adsorptive resins, can get purer Polysaccharides from Leaves of Moringa oleifera.Polysaccharides from Leaves of Moringa oleifera has significant protective effect to hepatic injury, is confirmed by animal experiment, and Polysaccharides from Leaves of Moringa oleifera is respectively provided with preventive and therapeutic effect to acute and subacute alcoholic liver injury.Polysaccharides from Leaves of Moringa oleifera can be applied well in exploitation preventing and treating alcoholic liver injury medicine and functional food.
Description
Technical field
The invention belongs to field of natural product chemistry, damage in preparation preventing and treating alcoholic liver particularly to a kind of Polysaccharides from Leaves of Moringa oleifera
Application in vulnerary thing and food.
Background technology
Alcoholic liver disease (Alcoholic Liver Disease, ALD) is that the alcoholic liver that long-term alcohol causes is damaged
Wound, the acute and chronic hepatic injury based on liver metabolism disorder.Clinically it is mainly shown as three kinds of forms：Alcoholic fatty liver,
Alcoholic hepatitis and alcoholic cirrhosises, these three forms can individually or be mixed.According to World Health Organization's statistics：The whole world
There are about 1500～20,000,000 people's excessive drinkings, wherein 10%～20% has different degrees of alcoholic liver disease.In American-European countries, Alcoholic
Hepatopathy is one of young and middle-aged main causes of death；North China's Epidemiological study in China showed from early 1980s
To the beginning of the nineties, ratio in population for the alcoholic rises to 14.3% from 0.21%；The beginning of this century, south and Midwest
Province Epidemiological study shows that the crowd of drinking increases to 30.9%～43.4%.Alcoholic liver disease accounts for same period hepatopathy inpatient
Ratio rises continuous, increases to 21.3% in 1996 from 4.2% in 1991.Especially in recent years, increase with China's economy
Long, people's social life mode development in pluralism, the total number of persons of the total amount of China's pre-capita consumption ethanol and consumption ethanol all exists
Rise year by year, alcohol abuse and alcohol dependence have become the whole world or even a Chinese public health problem becoming increasingly conspicuous, secondary
Alcoholic liver injury also become a very important disease in China.Therefore, find and research and development can effectively prevent and treat ethanol
The medicine of property hepatopathy and health food are very urgent.
At present, alcoholic liver injury temporarily no specific medicament and Therapeutic Method.In preventing and treating, still take and taken the photograph with reducing ethanol
Enter, the measure based on discovery early and early symptom management etc.；When obvious hepatic injury occurs then with liver protection and Supporting Therapy's phase
In conjunction with therapy, Drug therapy many from liver protection fibrosis, non-specific antiinflammatory, the aspect such as antioxidation is set about.Although currently having
The research report Chinese crude drug such as Radix Puerariae can be used for Antialcoholic liver-protecting, but most of Chinese crude drug toxic side effect, only can be used as medicine it is impossible to
It is eaten for a long time, as food；And most plants of report are all that liver-protecting efficacy composition is indefinite, with Plant crude extract
Use, limit the scope of use.So, find and become with using the single class having Antialcoholic liver-protecting effect in natural plant resource
Point, and developed into the medicine and health food extensive concern by medicine and food service industry.A class liver-protecting efficacy in plant
The discovery of composition is safe and effective, quality controllable for developing, and can effectively prevent and treat the health food of alcoholic liver injury, medicine has
Great meaning.
Moringa (Moringa oleifera Lam.), is Moringa suborder Moringaceae (Moringaceae) Moringa
(Moringa) plant, is perennial tropical and subtropical zone deciduous tree, Yin Qigen has acid, therefore thus Moringa of gaining the name.Moringa
Root, leaf, the equal edible of okra fruit, containing several mineral materials, vitamin, nutrient substance is very many；In addition, its root, skin, leaf, fruit
Medical material can be made, therefore Moringa has the good reputations such as " tree of miracle ", " diamond in plant ".Moringa has multiple medicinal valencys
Value, can treat the diseases such as ulcer, hypertension, hyperlipidemia, cancer, diabetes, inflammation, be the plant of great research and development potentiality
Thing.
Abroad the existing research with regard to Moringa antioxidation and liver protection reports, the scholar such as Sharifudin is with to acetyl at present
Amino phenols (APAP) inducing mouse produce hepatic injury, then with N-acetylcystein as control drug, have studied leaf of Moringa and
The therapeutical effect to mouse liver injury for the flower extract, result shows the Mouse Liver that leaf of Moringa and flower extract excessively cause to APAP
Damage has potential therapeutical effect (Sharifudin SA, Fakurazi S, Hidayat MT, et al.Therapeutic
potential of Moringa oleifera extracts against acetaminophen-induced
Hepatotoxicity in rats [J] .Pharmaceutical Biology, 2013,51 (3)：279-288).Sharma etc.
Have studied the protective effect to the acetsminophen that 7,12- dimethylbiphenyl [a] anthracene (DMBA) induces for the Moringa, find its energy
Reduce the liver aspartate transaminase (AST) of liver injury model mice, alanine aminotransferase (ALT) and the alkaline phosphorus of DMBA induction
Sour enzyme (ALP) level, the pathological change of the notable liver organization reversing DMBA induction.Result shows that Moringa induces to mice DMBA
Hepatocyte injury there is good liver protection and antioxidation potential (Sharma V, Paliwal R, Janmeda P, et
Al.Chemopreventive efficacy of Moringa oleifera pods against 7,12-
dimethylbenz[a]anthracene induced hepatic carcinogenesis in mice[J].Asian
Pacific Journal of Cancer Prevention, 2012,13 (6)：2563-2569).Sinha etc. have studied Moringa
The protective effect of the lipid peroxidation injury that leaf extract leads to radiation.Mice is exposed to 60 Co- after being administered 15 days
Gamma-radiation, observes the change such as liver lipid peroxidation, reduced glutathion of irradiation model mice, and result shows the leaf of Moringa
Extract can prevent hepatic lipid peroxidation to damage (Sinha M, Das DK, Datta S, et al.Amelioration of
ionizing radiation induced lipid peroxidation in mouse liver by Moringa
Oleifera Lam leaf extract [J] .Indian Journal of Experimental Biology, 2012,50
(3)：209-215).But in above research, leaf of Moringa or flower are used for resisting DMBA and APAP compound, and 60 Co- γ spokes
Penetrating the hepatic injury causing is all to take crude extract, does not damage with regard to a single class chemical composition such as Polysaccharides from Leaves of Moringa oleifera resisting alcoholic liver
The research of wound and application.
Content of the invention
In order to overcome shortcoming and the deficiency of above-mentioned prior art, the primary and foremost purpose of the present invention is to provide Polysaccharides from Leaves of Moringa oleifera to exist
Application in preparation preventing and treating alcoholic liver injury medicine and food.
The purpose of the present invention is realized by following proposal：
Application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera.
Described Polysaccharides from Leaves of Moringa oleifera is prepared by following methods：
(1) get the raw materials ready：Leaf of Moringa be dry, pulverize, cross 80 mesh sieves, obtain leaf of Moringa powder standby；
(2) defat：Take the leaf of Moringa powder of gained in step (1), add petroleum ether backflow defat, then filter and gained is filtered
Slag is dried, standby；
(3) extract：Take obtain in step (2) filtering residue is dried, with water as solvent, solid-liquid ratio is 1:15～1:25,60
DEG C～80 DEG C at extract 0.5～1.5h, extraction time be 3 times, merge Aqueous extracts, through 14000rpm centrifugation, take supernatant, obtain peppery
Wooden leaf crude polysaccharides solution；
(4) precipitate with ethanol：The leaf of Moringa crude polysaccharides solution obtaining in step (3) is concentrated into 1/2～1/3, is adjusted with dehydrated alcohol
Alcohol content, to 75%～85%, precipitates 12h, filters, takes precipitation, obtain leaf of Moringa crude polysaccharides；
(5) removing protein：The leaf of Moringa crude polysaccharides deionized water obtaining in step (4) is dissolved, adds polysaccharide liquid 1/5 body
Long-pending Sevag reagent (chloroform and n-butyl alcohol volume ratio 4: 1), magnetic agitation 30～35min after mixing, 4500rpm are centrifuged
20min, is processed 3 times repeatedly；
(6) purification：Product after removing protein in step (5) is added in adsorption resin column, carries out eluting with distilled water, wash
Separation of flow speed 2mL/min, eluent is concentrated into the 1/5 of original volume, obtains concentrated solution, concentrated solution is placed in 80 DEG C of water bath methods to phase
It is the extractum of 0.16～0.25 (with water as reference material) to density, be vacuum dried at 60 DEG C, obtain Polysaccharides from Leaves of Moringa oleifera powder.
In step (2), the amount of petroleum ether used is the corresponding petroleum ether using 3mL of leaf of Moringa powder of every 1g；Described returns
Stream defat refers to backflow twice, and each return time is 6h；The boiling range of petroleum ether used is 60 DEG C～90 DEG C.
Extraction conditions described in step (3) are preferably：Solid-liquid ratio is 1:20, extract 1h at 80 DEG C, extraction time is 3
Secondary.
Alcohol content described in step (4) is preferably 80%.
The consumption of the Sevag reagent described in step (5) is many for obtain after the dissolving of leaf of Moringa crude polysaccharides deionized water
The 1/5 of sugar liquid volume；Described Sevag reagent is 4 for volume ratio:1 chloroform and the mixed liquor of n-butyl alcohol.
Adsorption resin column described in step (6) is AB-8 type macroporous adsorptive resins；Described relative density is with water
For reference material, preferably 0.20.
The formulation ingredients of described preventing and treating alcoholic liver injury medicine and food all include Polysaccharides from Leaves of Moringa oleifera and country's license
The adjuvants such as the acceptable binding agent of food grade of interpolation, lubricant, flavoring agent.
Described acceptable adjuvant is preferably at least in Icing Sugar, Mannitol, Microcrystalline Cellulose, soluble starch etc.
Kind.
Described preventing and treating alcoholic liver injury medicine can be various dosage forms, such as granule, tablet, capsule, oral liquid,
Powder etc..
The mechanism of the present invention：
Polysaccharides from Leaves of Moringa oleifera, it is to malonaldehyde (MDA), triglyceride (TG) in Models of Acute Alcoholic Liver Injury mice serum
It is significantly reduced effect, and superoxide dismutase (SOD) and glutathion peroxidase (GSH-px enzyme) vigor are had
Potentiation, is conducive to mitigation, defence and the reparation of alcoholic liver injury.Polysaccharides from Leaves of Moringa oleifera can reduce subacute alcoholic liver to be damaged
Wound model mice serum low-density lipoprotein cholesterol (LDL-C), glutamic oxaloacetic transaminase, GOT (GOT), alanine aminotransferase
(GPT), T-CHOL (T-CHO), total bilirubin (T-BIL) content, makes hepatocyte function damage and mitigates and recover normal.Due to
Malonaldehyde (MDA) is one of most important product of membrane lipid peroxidatio, and its generation can cause biological big point of protein, nucleic acid etc.
The cross-linked polymeric of son, makes function damage or the forfeiture of film, produces cytotoxicity；Grind according to alcoholic liver injury pathogenesis
Study carefully discovery, alcoholic liver injury can cause content of triglyceride in hepatic tissue to build up, and cholesterol biosynthesis are strengthened, and causes corresponding load
The change of lipoprotein content, thus leading to hepatocellular function damage, and alcoholic liver steatosis and associated serum index
Change is the Early manifestation of alcoholic liver injury.Glutathion peroxidase (GSH-px) is the one kind being widely present in body
The enzyme that important catalyzing hydrogen peroxide decomposes.Its special catalysis reduced glutathion (GSH) is anti-to the reduction of hydrogen peroxide
Should, protection membrane structure and fully functional effect can be played.The active center of GSH-px is selenocystein, and selenium is
The required part of GSH-px, every mole enzyme contains 4 gram atom selenium.The vigor measuring GSH-px can be used as measurement body selenium level
A biochemical indicator.Glutathion peroxidase (GSH-px) can promote hydrogen peroxide (H2O2) and reduced form gluathione
Peptide (GSH) reaction generates H2O and oxidized form of glutathione (GSSG), the vigor of glutathion peroxidase can use enzymatic reaction
Speed to represent, measure the consumption of reduced glutathion in this enzymatic reaction, then can obtain the vigor of enzyme.Serum T-BIL
(total bilirubin, T-BIL) be bilirubin direct (direct bilirubin, DBIL) with unconjugated bilirubin (miL) it
Be heme catabolism product.The hemoglobin that serum bilirubin is mainly derived from red blood cell aging cracking and discharges, and liver
Dirty undertaking acts on to bilirubinic metabolite clearance.Therefore, hepatocyte is impaired or extrahepatic duct blocks etc., and reason can cause serum
T-BIL raises, and is the sensitive indicator in Liver function grade.Transaminase is requisite important thing in hepatocyte metabolic process
Matter, when hepatocyte is subject to various impacts to cause the pathology damage such as inflammation, necrosis, transaminase releasably enters blood, leads to serum
Transaminase raises, and serum transaminase is the important indicator of clinical reflection hepatocyte injury.
The present invention, with respect to prior art, has such advantages as and beneficial effect：
1st, the preparation method of the present invention can make leaf of Moringa crude polysaccharides yield increase, and after refinement treatment, energy
Obtain purer Polysaccharides from Leaves of Moringa oleifera.
2nd, the Polysaccharides from Leaves of Moringa oleifera of present invention preparation shows to acute and subacute alcoholic liver injury model experiment mice
Significant protective effect.Therefore, the Polysaccharides from Leaves of Moringa oleifera of present invention preparation can be applicable to anti-liver injury field especially alcoholic liver
Damage.
Brief description
Fig. 1 is embodiment 6 empty control group mice hepatic tissue section figure.
Fig. 2 is model group murine liver tissue slice map in embodiment 6.
Fig. 3 is Polysaccharides from Leaves of Moringa oleifera high dose group murine liver tissue slice map in embodiment 6.
Fig. 4 is Polysaccharides from Leaves of Moringa oleifera middle dose group murine liver tissue slice map in embodiment 6.
Fig. 5 is Polysaccharides from Leaves of Moringa oleifera low dose group murine liver tissue slice map in embodiment 6.
Specific embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention do not limit
In this.
Agents useful for same of the present invention all can be buied from market routine if no special instructions.
Embodiment 1：The preparation of Polysaccharides from Leaves of Moringa oleifera
(1) get the raw materials ready：Leaf of Moringa be dry, pulverize, cross 80 mesh sieves, obtain leaf of Moringa powder standby；
(2) defat：Take the leaf of Moringa powder obtaining in step (1), add the oil of 3 times of volumes (g: mL) of leaf of Moringa silty amount
Ether backflow defat 2 times, each 6h, filter, by filter residue and drying, standby；
(3) extract：Take obtain in step (2) filtering residue to be dried with water as solvent, solid-liquid ratio is 1:15, extract at 60 DEG C
0.5h, extraction time is 3 times, merges Aqueous extracts, through 14000rpm centrifugation, takes supernatant, obtains leaf of Moringa crude polysaccharides solution；
(4) precipitate with ethanol：The leaf of Moringa crude polysaccharides solution obtaining in step (3) is concentrated into 1/2-1/3, is adjusted with dehydrated alcohol and contain
Alcohol amount, to 75%, precipitates 12h, filters, takes precipitation, obtain leaf of Moringa crude polysaccharides；
(5) removing protein：The leaf of Moringa crude polysaccharides deionized water obtaining in step (4) is dissolved, adds polysaccharide liquid 1/5 body
Long-pending Sevag reagent (chloroform and n-butyl alcohol volume ratio 4: 1), magnetic agitation 30～35min after mixing, 4500rpm are centrifuged
20min, is processed 3 times repeatedly.
(6) purification：The Polysaccharides from Leaves of Moringa oleifera extracting solution obtaining in step (5) is added in AB-8 type macroporous adsorptive resins,
Carry out eluting, elution flow rate 2mL/min with distilled water, eluent is concentrated into the 1/5 of original volume, obtains concentrated solution, by concentrated solution
Being placed in 80 DEG C of water bath methods to relative density is the extractum of 0.16 (with water as reference material), is vacuum dried, obtains peppery at 60 DEG C
Wooden leaf polyose powder.
Embodiment 2：The preparation of Polysaccharides from Leaves of Moringa oleifera
(1) get the raw materials ready：Leaf of Moringa be dry, pulverize, cross 80 mesh sieves, obtain leaf of Moringa powder standby；
(2) defat：Take the leaf of Moringa powder in step (1), add the petroleum ether of 3 times of volumes (g: mL) of leaf of Moringa silty amount to return
Stream defat 2 times, each 6h, filter, by filter residue and drying, standby；
(3) extract：Take obtain in step (2) filtering residue to be dried with water as solvent, solid-liquid ratio is 1: 20, extract at 80 DEG C
1h, extraction time is 3 times, merges Aqueous extracts, through 14000rpm centrifugation, takes supernatant, obtains leaf of Moringa crude polysaccharides solution；
(4) precipitate with ethanol：The leaf of Moringa crude polysaccharides solution obtaining in step (3) is concentrated into 1/2-1/3, is adjusted with dehydrated alcohol and contain
Alcohol amount, to 80%, precipitates 12h, filters, takes precipitation, obtain leaf of Moringa crude polysaccharides；
(5) removing protein：The leaf of Moringa crude polysaccharides deionized water obtaining in step (4) is dissolved, adds polysaccharide liquid 1/5 body
Long-pending Sevag reagent (chloroform and n-butyl alcohol volume ratio 4: 1), magnetic agitation 30-35min after mixing, 4500rpm is centrifuged 20min,
Repeatedly process 3 times；
(6) purification：The Polysaccharides from Leaves of Moringa oleifera extracting solution obtaining in step (5) is added in AB-8 type macroporous adsorptive resins,
Carry out eluting, elution flow rate 2mL/min with distilled water, eluent is concentrated into the 1/5 of original volume, obtains concentrated solution, by concentrated solution
Being placed in 80 DEG C of water bath methods to relative density is the extractum of 0.20 (with water as reference material), is vacuum dried, obtains peppery at 60 DEG C
Wooden leaf polyose powder.
Embodiment 3：The preparation of Polysaccharides from Leaves of Moringa oleifera
(1) get the raw materials ready：Leaf of Moringa be dry, pulverize, cross 80 mesh sieves, obtain leaf of Moringa powder standby；
(2) defat：Take the leaf of Moringa powder in step (1), add the petroleum ether of 3 times of volumes (g: mL) of leaf of Moringa silty amount to return
Stream defat 2 times, each 6h, filter, by filter residue and drying, standby；
(3) extract：Take obtain in step (2) filtering residue to be dried with water as solvent, solid-liquid ratio is 1: 25, extract at 70 DEG C
1.5h, extraction time is 3 times, merges Aqueous extracts, through 14000rpm centrifugation, takes supernatant, obtains leaf of Moringa crude polysaccharides solution；
(4) precipitate with ethanol：The leaf of Moringa crude polysaccharides solution obtaining in step (3) is concentrated into 1/2-1/3, is adjusted with dehydrated alcohol and contain
Alcohol amount, to 85%, precipitates 12h, filtration, takes precipitation to obtain final product leaf of Moringa crude polysaccharides；
(5) removing protein：The leaf of Moringa crude polysaccharides deionized water obtaining in step (4) is dissolved, adds polysaccharide liquid 1/5 body
Long-pending Sevag reagent (chloroform and n-butyl alcohol volume ratio 4:1), magnetic agitation 30-35min after mixing, 4500rpm is centrifuged 20min,
Repeatedly process 3 times；
(6) purification：The Polysaccharides from Leaves of Moringa oleifera extracting solution obtaining in step (5) is added in AB-8 type macroporous adsorptive resins,
Carry out eluting, elution flow rate 2mL/min with distilled water, eluent is concentrated into the 1/5 of original volume, obtains concentrated solution, by concentrated solution
Being placed in 80 DEG C of water bath methods to relative density is the extractum of 0.25 (with the reference material of water), is vacuum dried, obtains peppery at 60 DEG C
Wooden leaf polyose powder.
Embodiment 4：The determination test of Polysaccharides from Leaves of Moringa oleifera content
1. the preparation of reference substance solution：Precision weighs the glucose 5mg being dried to constant weight, is settled to 50mL's with distilled water
In volumetric flask, shake up, concentration is 0.1mg/mL, then accurate absorption 1,2,3,4,5mL is put in 10mL volumetric flask respectively, to distill
Water dilution constant volume shakes up, and obtains 0.01mg/mL, the control series of 0.02mg/mL, 0.03mg/mL, 0.04mg/mL, 0.05mg/mL
Product solution.
2. the drafting of standard curve：The above-mentioned control series product solution 1mL of accurate absorption, is placed in 10mL tool plug test tube, ice
Water-bath 5min, adds 0.2% Anthrone Sulphuric acid test solution 4mL to shake up, and is immediately placed on boiling water bath heating 10min, and taking-up cold water cools down
To room temperature, room temperature places 10min, measures trap at 620nm, separately accurate absorption distilled water 1mL, is placed in 10mL tool plug test tube
In, ice-water bath 5min, add 0.2% sulphuric acid anthrone test solution 4mL to shake up, be immediately placed on boiling water bath heating 10min, taking-up cold water
It is cooled to room temperature, room temperature placement 10min about, as blank.With absorbance (A) as vertical coordinate, glucose control product are dense
Degree (C) is abscissa, obtains glucose standard curve.
3. measure：Weigh experimental example 1, experimental example 2, the Polysaccharides from Leaves of Moringa oleifera of experimental example 3 preparation respectively, prepared with distilled water
1mg/mL sample solution, takes 1mL to be placed in 10mL tool plug test tube, and ice-water bath 5min adds 0.2% sulphuric acid anthrone test solution 4mL to shake
Even, it is immediately placed on boiling water bath heating 10min, taking-up cold water is cooled to room temperature, and room temperature places 10min, measures and inhale at 620nm
Receipts degree, by concentration of glucose (C) in regression equation calculation test liquid.Result is as shown in table 1.
The determination test data of table 1 Polysaccharides from Leaves of Moringa oleifera content
From table 1 it follows that the Polysaccharides from Leaves of Moringa oleifera content highest of the method preparation of embodiment 2, compared with Example 3,
Solution usage is fewer, and extraction time is also comparatively short, and content is higher, is more suitably applied in production technology.
Embodiment 5：The impact test to Models of Acute Alcoholic Liver Injury mice for the Polysaccharides from Leaves of Moringa oleifera
1. material：
(1) laboratory animal：
Male mice 77,18～22 grams of weight, laboratory animal credit number are planted in SPF level Kunming (KM)：SCXK (Guangdong)
2013-0034, is provided by Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center.Using credit number：SYXK (Guangdong) 2014-0136.
(2) reagent：
Bifendate (Biphenyldicarboxylate, BP) drop pill, the limited public affairs of a Community in Baiyunshan, Guangzhou group of stars (Pharmaceutical) share
Department produces；The Polysaccharides from Leaves of Moringa oleifera preparing in the embodiment of the present invention 2.MDA, TG, SOD, GSH-px enzyme detection kit is by south
Capital is built up Bioengineering Research Institute and is provided.
2. laboratory animal packet：77 Male Kunming strain mice are randomly divided into 6 groups, respectively blank control group, model group,
Bifendate (15mg/mL) group, the high, medium and low dosage of Polysaccharides from Leaves of Moringa oleifera of the present invention (concentration be respectively 200mg/mL, 100mg/mL,
50mg/mL) group, wherein, blank control group 12, each 13 of remaining each group.
3. test method：
Experiment prospective adaptation is raised 5 days, free diet, the food ration of daily observed and recorded each group mice and amount of drinking water.Real
Test first 16 days, each group mouse stomach gives the tested material of various dose, except blank control group and model group mouse stomach give
Outside amount normal saline, each dosage Polysaccharides from Leaves of Moringa oleifera group mice gives 2 tested materials daily, fills by weight 0.12mL/10g dosage
Stomach, bifendate group mice press people's daily quantity conversion after dosage medication, i.e. weight 0.12mL/10g dosage gavage,
One time a day.Weigh weekly each group Mouse Weight twice, to adjust corresponding tested material consumption.Test the 17th day, start modeling, remove
Outside blank control group, remaining each group mice gavages 50% food grade ethanol 1 time by weight 0.1mL/10g dosage daily, continuously
Give 9 days ethanol, tested material give dosage and method is the same.Modeling terminates rear each group mice and gives corresponding tested material 3～5 again
My god.Before experiment terminates, fasting 16h after last gavage, weigh body weight, then pluck eyeball and take blood, by the whole blood of collection 20
After placing 1h at DEG C, it is centrifuged 10min through rotating speed 3 000rpm, draw serum in sterilizing 1.5mL centrifuge tube, be placed in 20 DEG C of ice
Case saves backup.Cervical dislocation puts to death mice, weighs mice organs, liver is placed on ice platform, is quickly cut with knife blade
Liver lobus dexter, notes taking right lobe of liver same area hepatic tissue to be divided into 2～3 parts and be distributed into clean cryopreservation tube and is placed in -80 DEG C of ice
Save backup in case.
4. Testing index and detection method
4.1 Triglycerides in Serums (TG), the mensure of malonaldehyde (MDA) content.In strict accordance with kit specification operation
Step is carried out.
Malonaldehyde (MDA) content, superoxide dismutase (SOD) activity, Glutathione thing peroxidase in 4.2 hepatic tissues
(GSH-px) mensure of activity.Take a liver organization of above-mentioned preservation, be configured to 10% liver homogenate with cold saline, press
Kit specification operating procedure operates, MDA content, SOD activity, GSH-px activity in detection liver.
4.3 statistical analysis
Experimental data all carries out statistical disposition using SPSS 15.0 software kit, and result is represented with x ± s, compares employing between group
One factor analysis of variance, P ＜ 0.05 indicates significant difference.
5. result of the test：Take blood, separate serum, measure malonaldehyde (MDA), triglyceride (TG) content in mice serum；
Liver organization detection MDA, superoxide dismutase (SOD enzyme) vigor and glutathion peroxidase (GSH-px enzyme) is taken to live
Power.Result is as shown in table 2～3.
The impact to acute alcohol-induced hepatic injury mice serum TG and MDA content for table 2 Polysaccharides from Leaves of Moringa oleifera
Note：Compared with model group, * represents P<0.05, * * represents P<0.01.
Drawn by table 2, compare with blank control group mice, malonaldehyde (MDA) and triglyceride in model group mice serum
(TG) content is significantly raised, has significant significant difference (P<0.05,P<0.01), illustrate that ethanol inducing acute hepatic injury is little
Mouse model is successfully established.Compared with model group, the Polysaccharides from Leaves of Moringa oleifera high, medium and low dosage group mice serum triglyceride of the present invention
(TG) it is decreased obviously (P<0.05, P<0.01), decline the most notable, positive control drug bifendate group mice blood with high dose group
Clear TG also (P lower than model group mice serum TG level<0.05)；In terms of each group mice serum MDA detection level, model group mice
Serum is apparently higher than blank control group (P<0.05)；Compared with model group, Polysaccharides from Leaves of Moringa oleifera middle dose group mice serum MDA content
(8.139 ± 3.764nmol/ml) is less than model group mice MDA (11.511 ± 2.015nmol/ml) content (P<0.05), Moringa
Leaf polyose high and low dose group has the trend of reduction.Therefore, the Polysaccharides from Leaves of Moringa oleifera of the present invention is little to acute alcohol-induced hepatic injury experiment
Mouse model shows obvious hepatoprotective effect.
Table 3 Polysaccharides from Leaves of Moringa oleifera affects on acute alcohol-induced hepatic injury murine liver tissue MDA, SOD and GSH-px
Note：Compared with model group, * represents P<0.05, * * represents P<0.01.
Drawn by table 3, compare with blank control group mice, in model group murine liver tissue, malonaldehyde (MDA) content is obvious
Raise, there is significant significant difference (P<0.01), and SOD enzyme and GSH-px enzymatic activity are significantly lower than blank control group (P<
0.05).Process after alcoholic liver injury model mice it has been found that Polysaccharides from Leaves of Moringa oleifera through the high, medium and low dosage of Polysaccharides from Leaves of Moringa oleifera
Each dosage group murine liver tissue MDA level is all extremely remarkably decreased (P<0.01), in addition high dose group be less than blank control group little
Mus MDA level, the MDA level of bifendate group is also decreased obviously (P<0.05).From hepatic tissue SOD enzyme and GSH-px enzymatic activity
See, Polysaccharides from Leaves of Moringa oleifera high, medium and low dosage group Mouse Liver GSH-px enzymatic activity all substantially rises (P<0.05), Polysaccharides from Leaves of Moringa oleifera
The performance of low dose group Mouse Liver SOD enzyme activity significantly increases (P<0.01).Therefore, compared with model group, the leaf of Moringa of the present invention
Polysaccharide is significantly reduced effect (P to liver MDA<0.01), there is potentiation (P to SOD and GSH-px enzyme activity<0.05, P<
0.01), be conducive to improving the hepatic injury of model mice, Polysaccharides from Leaves of Moringa oleifera has defencive function to act on to acute alcohol-induced hepatic injury.
Embodiment 6：The impact test to subacute alcoholic liver injury model mice for the Polysaccharides from Leaves of Moringa oleifera
1. material：
(1) laboratory animal：
Male mice 80,18～22 grams of weight, laboratory animal credit number are planted in SPF level Kunming (KM)：SCXK (Guangdong)
2013-0034, is provided by Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center.Using credit number：SYXK (Guangdong) 2014-0136.
(2) reagent：
The Polysaccharides from Leaves of Moringa oleifera preparing in the embodiment of the present invention 2；Sunflower liver-protecting tablet, is had by Heilongjiang Kuihua Yaoye Co.Ltd
Limit company produces, lot number：Z20003336；MDA, GOT, GPT, T-CHO, LDL-C and T-BIL detection kit is built up by Nanjing
Bioengineering Research Institute provides.
2. laboratory animal packet：80 Male Kunming strain mice are randomly divided into 6 groups, respectively blank control group, model group,
(concentration is respectively 200mg/mL, 100mg/ for sunflower liver-protecting tablet (86mg/mL) group, the high, medium and low dosage of Polysaccharides from Leaves of Moringa oleifera of the present invention
ML, 50mg/mL) group, wherein, and blank control group 10, each 14 of remaining each group.
3. test method：
Experiment prospective adaptation is raised 5 days, free diet.Each group its mouse oral gavage gives different tested materials, except blank
Matched group and model group mouse stomach give outside normal saline, each Polysaccharides from Leaves of Moringa oleifera group mice give daily 2 times accordingly dense
The Polysaccharides from Leaves of Moringa oleifera of degree, by weight 0.12mL/10g dosage gavage, sunflower liver-protecting tablet group mice is pressed the daily quantity of people and rolls over
Dosage medication after calculation, i.e. weight 0.12mL/10g dosage gavage, one time a day, continuously gives 45 days.Mice weighs body weekly
Weight twice, and adjusts given the test agent dosage by body weight.In addition to blank control group, each group terminates first 14 days in experiment, presses daily
The oral gavage of 10mL/kg bw gives 30% food grade ethanol.Gavage time interval 4h of tested material and ethanol.Before experiment terminates
Fasting 12h, weighs body weight, then plucks eyeball blood sampling.The whole blood of collection is placed after 1h at 20 DEG C, through rotating speed 3 000rpmin
Centrifugation 10min, draws serum in sterilizing 1.5mL centrifuge tube, is placed in 20 DEG C of Refrigerator stores standby.Mice cervical dislocation is put to death,
Weigh mice organs, liver is placed on ice platform, quickly cuts liver lobus dexter with knife blade, note taking right lobe of liver same area
Hepatic tissue is used for pathological examination, puts into 10% neutral formalin solution and fixes, remaining hepatic tissue is divided into 2～3 parts points
Load clean cryopreservation tube and be placed in -80 DEG C of refrigerators and save backup.
4. Testing index and detection method
Low-density lipoprotein cholesterol (LDL-C), glutamic oxaloacetic transaminase, GOT (GOT), alanine aminotransferase in 4.1 serum
(GPT), T-CHOL (T-CHO), the mensure of total bilirubin (T-BIL) content.In strict accordance with kit specification operating procedure
Carry out.
The mensure of malonaldehyde (MDA) content in 4.2 hepatic tissues.Take a liver organization of above-mentioned preservation, use cold physiology salt
Water is configured to 10% liver homogenate, by the operation of kit specification operating procedure, MDA content in detection liver.
4.3 liver histopathology inspections：Using HE dyeing, with optical microscope, amplification is 400 × inspection Mouse Liver
The pathological section of tissue, observes hepatocellular degeneration (steatosis, hydropic degeneration, endochylema cohesion, balloon sample become), hepatic necrosis
Situations such as change with inflammation, and report is made to each group murine liver tissue pathological examination results.
4.4 statistical analysis
Data statistic analysis are carried out using SPSS 20.0 software.To between mice serum biochemical indicator, multigroup group of liver index
Mean compares using one factor analysis of variance (ANOVA), is checked with LSDL, S-N-K (S), during heterogeneity of variance during homogeneity of variance
Using Tamhane ' s T2 (M) and Dunnett ' s T3 inspection, the relatively significance of difference of each class mean, P ＜ 0.05 indicates
Significant difference.
5. result of the test：
5.1 biochemistry detection result：Take blood, separate serum, measure LDL-C, GOT, GPT, T-CHO, T-BIL in mice serum
Content；Take liver organization detection MDA level.According to SFDA《Health care test evaluation technical specification》In " to chemical liver injury
There is assistant protection function evaluation methodology (revised draft)》" related criterion, experimental result is judged.Result such as table 4～5
Shown.
The impact to biochemical indicator content each in mice serum for table 4 Polysaccharides from Leaves of Moringa oleifera
Note：Compared with model group, * represents P<0.05, * * represents P<0.01.
Drawn by table 4, subacute alcoholic hepatic injury model group mice serum LDL-C, GOT, GPT, T-CHO and T-BIL contain
Amount, obviously higher than blank control group, has notable significant difference (P ＜ 0.05, P ＜ 0.01), shows subacute alcoholic liver
Damage Establishment of mouse model success.
In addition to serum T-CHO level, Polysaccharides from Leaves of Moringa oleifera each dosage group mice serum LDL-C, GOT, GPT, T-CHO, T-BIL
Content is compared with model group mice, has different degrees of reduction (P ＜ 0.05).Polysaccharides from Leaves of Moringa oleifera damages to subacute alcoholic liver
Wound has defencive function to act on.
The impact to murine liver tissue malonaldehyde (MDA) content for table 5 Polysaccharides from Leaves of Moringa oleifera
Note：* is represented and is compared with model group, P ＜ 0.01.
Drawn by table 5, subacute alcoholic hepatic injury model group mice MDA content (0.422 ± 0.49 μm of ol/g) is significantly high
In blank control group (0.268 ± 0.29 μm of ol/g), difference has statistical significance (P ＜ 0.01).Polysaccharides from Leaves of Moringa oleifera is high, in,
Low dose group murine liver tissue MDA level is all substantially less than model group mice (0.422 ± 0.49 μm of ol/g), has statistics poor
Different (P ＜ 0.01), points out after giving Polysaccharides from Leaves of Moringa oleifera intervention, the biochemical indicator of model mice abnormal liver function is restored,
Hepatocyte function damages and mitigates.
The impact that 5.2 Polysaccharides from Leaves of Moringa oleiferas change to subacute alcoholic hepatic injury model mice hepatic pathology
To above-mentioned blank control group, model group, Polysaccharides from Leaves of Moringa oleifera high dose group, Polysaccharides from Leaves of Moringa oleifera middle dose group, leaf of Moringa
The murine liver tissue of polysaccharide low dose group carries out sections observation, and result such as Fig. 1 respectively, shown in Fig. 2, Fig. 3, Fig. 4, Fig. 5.
Wherein Fig. 1 is blank control group murine liver tissue slice map, from figure 1 it appears that the arrangement of liver cell rope is whole
Together, cell is in polygon, and it is clear to be arranged radially centered on central vein and demarcate, liver cell nuclear circular it is seen that double-core,
The phenomenons such as liver organization structural integrity, no edema and steatosis.
Fig. 2 is model group murine liver tissue slice map, from figure 2 it can be seen that lobuli hepatis structure is abnormal, hepatocyte arranges
Disorder, the universal swelling and degeneration of hepatocyte and necrosis.The hepatocyte of necrosis is in that disperse shape is distributed, and centrilobular vein area hepatocyte is bad
Dead degree is the heaviest, and liver interstitial has severe congestion.
Fig. 3 is Polysaccharides from Leaves of Moringa oleifera high dose group murine liver tissue slice map, from figure 3, it can be seen that hepatic tissue structure is different
Chang Chengdu substantially changes, and lobuli hepatis structure gradually recovers normal, and hepatocyte arrangement is in streak, the hepatocyte number of degeneration and necrosis
Amount reduces, the increasing of hepatocyte double-core, and illustrate that hepatocyte function strengthens, and hepatocyte is divided and increases, hepatic sinusoid, central veins of liver and
Liver interstitial congestion degree significantly mitigates.
Fig. 4 is Polysaccharides from Leaves of Moringa oleifera middle dose group murine liver tissue slice map, figure 4, it is seen that hepatic tissue structure changes
Become, hepatocellular degeneration and necrosis are distributed in stove shape, and degree relatively low-dose group is light, and the hepatocyte in central veins of liver area just recovers first
Often, the secondary hepatocyte for lobules of liver limiting plate area, the congestion degree of hepatic sinusoid and liver interstitial substantially reduces, a large amount of cell infiltration.
Fig. 5 is Polysaccharides from Leaves of Moringa oleifera low dose group murine liver tissue slice map, from figure 5 it can be seen that hepatic tissue structure has
Pathologic changes, and hepatic necrosis is in stove shape, and the hepatocyte quantity of necrosis is many compared with middle dose group, and lesion degree is also compared with middle dose group
Weight, the inflammatory trifle that the visible inflammatory cell hypertrophy of section is formed.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not subject to above-described embodiment
Limit, other any spirit without departing from the present invention and the change made under principle, modification, replacement, combine, simplify,
All should be equivalent substitute mode, be included within protection scope of the present invention.
Claims (10)
1. application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera.
2. application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera according to claim 1,
It is characterized in that described Polysaccharides from Leaves of Moringa oleifera is prepared by following methods：
(1) get the raw materials ready：Leaf of Moringa be dry, pulverize, cross 80 mesh sieves, obtain leaf of Moringa powder standby；
(2) defat：Take the leaf of Moringa powder of gained in step (1), add petroleum ether backflow defat, then filter and gained filtering residue is done
Dry, standby；
(3) extract：Take obtain in step (2) filtering residue is dried, with water as solvent, solid-liquid ratio is 1:15～1:25,60～80
Extract 0.5～1.5h at DEG C, extraction time is 3 times, merges Aqueous extracts, through 14000rpm centrifugation, take supernatant, obtain leaf of Moringa thick
Polysaccharide solution；
(4) precipitate with ethanol：The leaf of Moringa crude polysaccharides solution obtaining in step (3) is concentrated into 1/2～1/3, adjusts containing alcohol with dehydrated alcohol
Measure to 75%～85%, precipitate 12h, filter, take precipitation, obtain leaf of Moringa crude polysaccharides；
(5) removing protein：The leaf of Moringa crude polysaccharides deionized water obtaining in step (4) is dissolved, adds Sevag reagent, mixing
Magnetic agitation 30～35min afterwards, 4500rpm are centrifuged 20min, repeatedly process 3 times；
(6) purification：Product after removing protein in step (5) is added in adsorption resin column, carries out eluting, eluting stream with distilled water
Fast 2mL/min, eluent is concentrated into the 1/5 of original volume, obtains concentrated solution, and concentrated solution is placed in 80 DEG C of water bath methods to relatively close
Spend the extractum for 0.16～0.25, be vacuum dried at 60 DEG C, obtain Polysaccharides from Leaves of Moringa oleifera powder.
3. application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera according to claim 2,
It is characterized in that：
In step (2), the amount of petroleum ether used is the corresponding petroleum ether using 3mL of leaf of Moringa powder of every 1g；Described backflow takes off
Fat refers to backflow twice, and each return time is 6h；The boiling range of petroleum ether used is 60 DEG C～90 DEG C.
4. application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera according to claim 2,
It is characterized in that：
Sevag reagent described in step (5) is 4 for volume ratio:1 chloroform and the mixed liquor of n-butyl alcohol；Described Sevag examination
The consumption of agent is the 1/5 of the polysaccharide liquid volume obtaining after leaf of Moringa crude polysaccharides deionized water dissolves.
5. application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera according to claim 2,
It is characterized in that：
Adsorption resin column described in step (6) is AB-8 type macroporous adsorptive resins.
6. application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera according to claim 2,
It is characterized in that：
Relative density described in step (6) is with water as reference material.
7. application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera according to claim 2,
It is characterized in that：
Extraction conditions described in step (3) are：Solid-liquid ratio is 1: 20, extracts 1h at 80 DEG C, and extraction time is 3 times；
Alcohol content described in step (4) is 80%；
Relative density described in step (6) is 0.20.
8. application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera according to claim 2,
It is characterized in that：
The formulation ingredients of described preventing and treating alcoholic liver injury medicine and food all include Polysaccharides from Leaves of Moringa oleifera and country's license is added
The acceptable adjuvant of food grade.
9. application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera according to claim 8,
It is characterized in that：
Described acceptable adjuvant is Icing Sugar, Mannitol, Microcrystalline Cellulose, at least one in soluble starch.
10. application in preparation preventing and treating alcoholic liver injury medicine and food for the Polysaccharides from Leaves of Moringa oleifera according to claim 8,
It is characterized in that：
Described preventing and treating alcoholic liver injury medicine is granule, tablet, capsule, oral liquid or powder.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107233366A (en) * | 2017-05-17 | 2017-10-10 | 华南农业大学 | A kind of leaf of Moringa granule and preparation method and application |
| CN107573439A (en) * | 2017-11-03 | 2018-01-12 | 隆安县荣胜养殖有限公司 | A kind of technique of extraction purification Moringa polysaccharide |
| CN108354173A (en) * | 2018-04-11 | 2018-08-03 | 钦州学院 | A kind of Moringa selenium-rich polysaccharide buccal tablet |
| CN109422818A (en) * | 2017-08-21 | 2019-03-05 | 华南农业大学 | A kind of preparation method and applications of Polysaccharides from Leaves of Moringa oleifera MLP20-1 |
| KR20190108949A (en) * | 2018-03-16 | 2019-09-25 | (주)마성 | Process for preparing alcoholic fatty liver relaxation composition using Moringa oleifera and composition thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20150041534A (en) * | 2013-10-08 | 2015-04-16 | 주식회사 강남식품 | Reformed rice comprising powder of Moringa and mothod for preparing the same |
| CN104829743A (en) * | 2015-05-25 | 2015-08-12 | 萧丽雅 | Preparation method and use of moringa oleifera leaf polysaccharides |
| CN104938900A (en) * | 2015-07-24 | 2015-09-30 | 海南木辣达生物科技有限公司 | Moringa hangover-alleviating composition and preparation method thereof |
| CN105311069A (en) * | 2014-08-01 | 2016-02-10 | 薛永新 | Traditional Chinese medicine composition for reducing blood lipid and protecting livers |
| CN105614851A (en) * | 2015-12-27 | 2016-06-01 | 劲膳美生物科技股份有限公司 | Medical formula food for hepatic encephalopathy |
-
2016
- 2016-11-29 CN CN201611071660.4A patent/CN106474145B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20150041534A (en) * | 2013-10-08 | 2015-04-16 | 주식회사 강남식품 | Reformed rice comprising powder of Moringa and mothod for preparing the same |
| CN105311069A (en) * | 2014-08-01 | 2016-02-10 | 薛永新 | Traditional Chinese medicine composition for reducing blood lipid and protecting livers |
| CN104829743A (en) * | 2015-05-25 | 2015-08-12 | 萧丽雅 | Preparation method and use of moringa oleifera leaf polysaccharides |
| CN104938900A (en) * | 2015-07-24 | 2015-09-30 | 海南木辣达生物科技有限公司 | Moringa hangover-alleviating composition and preparation method thereof |
| CN105614851A (en) * | 2015-12-27 | 2016-06-01 | 劲膳美生物科技股份有限公司 | Medical formula food for hepatic encephalopathy |
Non-Patent Citations (1)
| Title |
|---|
| 贺艳培 等: "辣木的研究进展", 《科学观察》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107233366A (en) * | 2017-05-17 | 2017-10-10 | 华南农业大学 | A kind of leaf of Moringa granule and preparation method and application |
| CN109422818A (en) * | 2017-08-21 | 2019-03-05 | 华南农业大学 | A kind of preparation method and applications of Polysaccharides from Leaves of Moringa oleifera MLP20-1 |
| CN107573439A (en) * | 2017-11-03 | 2018-01-12 | 隆安县荣胜养殖有限公司 | A kind of technique of extraction purification Moringa polysaccharide |
| KR20190108949A (en) * | 2018-03-16 | 2019-09-25 | (주)마성 | Process for preparing alcoholic fatty liver relaxation composition using Moringa oleifera and composition thereof |
| KR102183670B1 (en) * | 2018-03-16 | 2020-11-26 | (주)마성 | Process for preparing alcoholic fatty liver relaxation composition using Moringa oleifera and composition thereof |
| CN108354173A (en) * | 2018-04-11 | 2018-08-03 | 钦州学院 | A kind of Moringa selenium-rich polysaccharide buccal tablet |
| CN108354173B (en) * | 2018-04-11 | 2021-11-30 | 钦州学院 | Moringa oleifera selenium-rich polysaccharide buccal tablet |
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