CN101914164B - Application of polysaccharide from mulberry leaves in preventing and treating diabetes - Google Patents
Application of polysaccharide from mulberry leaves in preventing and treating diabetes Download PDFInfo
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- CN101914164B CN101914164B CN 201010246917 CN201010246917A CN101914164B CN 101914164 B CN101914164 B CN 101914164B CN 201010246917 CN201010246917 CN 201010246917 CN 201010246917 A CN201010246917 A CN 201010246917A CN 101914164 B CN101914164 B CN 101914164B
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Abstract
The invention relates to application of polysaccharide extracted from mulberry leaves in preventing and treating diabetes, belonging to the natural medicine application field. The application method comprises the following steps: crushing 10kg of the dry mulberry leaves, and adding 180L of distilled water to the crushed mulberry leaves; heating and then carrying out reflux extraction on the obtained mixture at the temperature of 70 DEG C for 90min; filtering the obtained extracting solution twice with three-layer gauze and then filtering with a ceramic membrane; decompressing and concentrating the obtained filtrate into 10L, and then deproteinizing the obtained concentrated solution with 10% of trichloroacetic acid; and placing the concentrated solution on an SEPHADEX G-10 column for separating and collecting components containing the polysaccharide from the mulberry leaves, concentrating the solution into 1L, and then placing the concentrated solution on a silicagel column for separating and collecting the components containing the polysaccharide from the mulberry leaves again to obtain the polysaccharide from the mulberry leaves, wherein, content of the polysaccharide from the mulberry leaves is 95.5%.
Description
Affiliated technical field:
The present invention relates to a kind of application of polysaccharides of Folium Mori in prevention and treatment diabetes of from mulberry leaf, extracting, belong to the technical field that natural drug is used.
Background technology
So far, global diabetic subject has reached 1.3 hundred million, and number is also increasing severely.Diabetes have become No. three killer after cardiovascular, tumour, are one of the world's three big bus health difficult problems.According to Ministry of Health's publish data, 2009 are only the city diabetic subject has surpassed 4,100 ten thousand, and newly-increased patient approximately 1,500,000 every year on average.Diabetes are the common disease that is caused by the h and E factor interaction, and are clinical take hyperglycemia as item key, and common sympton has many drinks, diuresis
Eat more and become thin etc.Diabetes can cause the infringement of health multisystem, cause that the absolute or relative hyposecretion of Regular Insulin and target tissue cell reduce insulin sensitivity, cause a series of metabolism disorder syndromes such as protein, fat, power and water Xie Zhi, wherein a hyperglycemia is item key.Also do not have at present specific medicine can give effective radical cure.From existing screening bioactive compounds, study the herbal medicine efficacy composition of known curative effect, have very great help to promoting the internationalization of tcm tool.
In the past, the treatment of diabetes mainly contained four kinds of methods: dietetic treatment, pharmacological agent, exercise therapy.Abroad, some famous physicians advocate that the diabetic subject should do suitable physical exertion, and physical exertion, dietary control, insulin injection are classified as the three magic weapons for the treatment of diabetes.In recent years, utilize TCM Treatment of Diabetes to become the hot subject of research.
Polysaccharides of Folium Mori is a kind of acid heteroglycan, mainly is comprised of uronic acid and various neutral sugar.Mulberry leaf different name Herba adianti myriosori is the Moraceae fallen leaves property leaf of grass mulberry tree for many years.Modern pharmacology and clinical study show, that the polysaccharide of one of mulberry leaf activeconstituents has is hypoglycemic, hypotensive, immunomodulatory, the multiple physiologically active such as antibiotic and antiviral.Especially Japan is more to the effective constituent research in the mulberry leaf both at home and abroad at present, finds that polysaccharide has hypoglycemic effect, and polysaccharide structures is analyzed.Therefore be subject to the attention on biomedical boundary.
Polysaccharide be a class by the biological response modifier of active research, the active polysaccharide that contains in the mulberry leaf has and strengthens immunologic function and anti-infectious effect, treats in addition the effect of hypercholesterolemia, and can strengthen blood vessel and prevention of arterial sclerosis, thrombosis, scheming infarction etc.The polysaccharides of Folium Mori nontoxicity has obvious health-care effect, especially has the immunocompetence function and more comes into one's own, and the polysaccharides of Folium Mori protective foods will become important a member of novel healthy food.Therefore the mulberry leaf resource of developing us just has very important effect.
China's mulberry tree kind, kind and mulberry fruit output all occupy first place, the world, and except the Qinghai-Tibet Platean, all there is cultivation all parts of the country.Most area is with the feeding cultivation of Sang Zuowei at present, and annual except raising silkworm, a lot of positions of mulberry tree are not all utilized.Although many researchs have reported that mulberry has multiple pharmacological effect, the research of anti-allergic effects also rarely has report.Document up to now focuses on that all the 1-DNJ that extracts in the mulberry leaf studies in the treatment diabetes, its mechanism of action in the treatment diabetes is fully aware of.We find the high 1-DNJ of the low 1-DNJ ratio content of content to the better effects if for the treatment of diabetes in research process, By consulting literatures is found polysaccharides of Folium Mori in not research aspect prevention and the treatment diabetes simultaneously.And polysaccharides of Folium Mori content in mulberry leaf is higher, draws materials conveniently, and is widely distributed, low price.Thereby, if polysaccharides of Folium Mori has certain effect at tool aspect prevention and the treatment diabetes, will mean a great to the Sang Jinhang deep processing.
Summary of the invention
The polysaccharides of Folium Mori that the present invention mentions is to distill hydro-thermal extraction, filtration, concentrating under reduced pressure, trichoroacetic acid(TCA) except albumen from drying mulberry leaf later through using, and then with post absorption, elutriant is concentrated, concentrating under reduced pressure makes the polysaccharides of Folium Mori sterling.Its polysaccharide content is more than 95%.
Technique effect:
1, used raw material, equipment is common common raw material, equipment in the inventive method, has avoided in the commercial process having reduced widely production cost for the dependence of expensive raw materials, instrument.
2, mulberry tree plantation easily, cuttage, seed broadcast all can, output is high, price is low, convenient sources, follow-up resource is secure.Compare with traditional extraction technology, this technology can shorten extraction time greatly, reduce energy consumption, reduce the stripping of impurity component.
3, the inventive method is simple to operate, does not need precision instrument or automatic equipment, can produce in the resourceful township and village enterprises of mulberry leaf, and this greatly reduces the production cost of polysaccharides of Folium Mori, has simplified production process.
4, polysaccharides of Folium Mori of the present invention has many-sided opsonization, and side effect is little, compares with chemical synthetic drug, without obvious dependency.
Embodiment
Below, the present invention will be further detailed with embodiment, but it is not limited to any or the similar example of these embodiment.
Embodiment 1: the preparation method of polysaccharides of Folium Mori
Pulverize dry mulberry leaf 10kg, add 180 L distilled water, 70 ℃ of heating and refluxing extraction 90min, three layers of filtered through gauze twice are used ceramic membrane filter again, are evaporated to 10 L, then concentrated solution removes albumen with 10% trichoroacetic acid(TCA), upper SEPHADEX G-10 post separated and collected contains the polysaccharides of Folium Mori component, is concentrated into 1L and goes up silicagel column separated and collected polysaccharides of Folium Mori composition again and be concentrated into driedly, obtains polysaccharides of Folium Mori.Measure through anthrone-sulphate method, wherein polysaccharides of Folium Mori content is 95.5%.
Embodiment 2: the mouse of polysaccharides of Folium Mori is fed experiment
1, Model of diabetic rat
(1) adult kunming mice (20-25g) cultivates under (25 ± 1 ° of C) ventilation condition, illumination 12h, and dark 12h, free choice feeding, standard are fed.
(2) diabetic mice is induced: according to the requirement of 45 mg/kg, by abdominal injection U-9889 (being dissolved in 0.1M citrate buffer solution PH4.5).Continuous 7 days, on an empty stomach mouse tail blood sampling selected the mouse of fasting plasma glucose 200 mg/dl as experimental subjects.2, experimental design:
2, experimental mouse is divided into 6 groups, 6 every group: test the line space abdomen blood sugar detection that advances
The 1st group: normal non-diabetic mouse, drink distilled water;
The 2nd group: diabetic mice, distilled water;
The 3-5 group: diabetic mice is given respectively Folium Mori extract 0.25,0.5,1.0 g/kg/day;
The 6th group: diabetic mice injection 4U/kg/day Regular Insulin continued for 6 weeks.
All experimental mouse use Instrument for Measuring Blood Sugar to measure blood sugar before experiment and the 6th all laggard line space abdomen blood sugar detections.During the fasting, stop to all foods of mouse except water.After initial period (before inducing diabetes) and 6 all treatments phases, caudal vein is got hematometry mda, lipid peroxide and glycolated hemoglobin.
3, use the production of thiobarbituric acid reactive method to detect MDA: blood plasma and thiobarbituric acid reaction generate coloured production of thiobarbituric acid reactive (describing according to Draper), measure under the 532nm wavelength, blood plasma MDA content uses the tetraethoxypropane typical curve.
4, Determination of lipid peroxide adopts the chemical examination of ferrous ion xylenol orange: Lipoperoxide thing Oxidation of Fe
2+To Fe
3+Bonding xylenol orange (describing according to Nourooz-Zadeh).Reactant is spectrophotometric determination under the 560nm wavelength.Lipid peroxide contents is used the hydrogen peroxide standard curve determination.Measure in the body Folium Mori extract to the effect of saccharification level by the method for chemical examination glycolated hemoglobin.The total antithrombotics of blood is used for measuring protoheme and glycolated hemoglobin concentration.The protoheme that colorimetric method for determining is total uses the turbid rate method of immune transmittance (COBAS INTEGRA 700 test kits) to measure glycolated hemoglobin.The glycolated hemoglobin amount represents with the % content of protoheme.
5, results of animal
(1) the mulberry leaf extracting solution is on the impact of tissue of experimental diabetic mice blood sugar
With diabetic model group relatively, mulberry leaf extracting solution group whole blood blood sugar obviously descends (P＜0.05), more experimental mouse is drunk, is eaten more, the diuresis symptom is obviously improved.But blood sugar and control group are higher, do not drop to normal level, and to play hypoglycemic effect more obvious along with the increase of Folium Mori extract concentration within the specific limits; But its effect does not have the obvious of Regular Insulin.Blood sugar no significant difference before and after the vacation of diabetic model group experimental mouse is raised (P〉0.05).The results are shown in Table 1:
Table 1 mulberry leaf extracting solution group is on the impact of artificial diabetes experimental mouse blood sugar
Table 1 The effects of extract of mulberry leaves on blood sugar level in diabetic rats (Jian x ± s, mmol/L)
(2) the mulberry leaf extracting solution is on the impact of tissue of experimental diabetic mice activity of plasma SOD
Diabetic groups experimental mouse SOD activity after the modeling obviously reduces than the SOD of control group experimental mouse is active, gives behind the mulberry leaf extracting solution Blood SOD Activity obviously raise (P＜0.05).The amount difference of giving of its Folium Mori extract also has this different effect.The results are shown in Table 2:
Table 2 mulberry leaf extracting solution group is on the impact of artificial diabetes experimental mouse plasma SOD activity
Table2 The effects of extract of mulberry leaves on SOD in plasma of diabetic rats (Jian x ± s, U/mL)
Model of diabetic rat SOD level is lower than normal experimental mouse SOD level as can be seen from Table 2, give the mulberry leaf extracting solution after, the activity of plasma SOD level of experimental mouse obviously raise (P＜0.05).Its active size that raises within the specific limits along with Folium Mori extract to measuring into the forward dependency, the SOD activity of Folium Mori extract group 3 has reached normal level.The injection of insulin group then so that its SOD activation recovering to normal level.
(3) the mulberry leaf extracting solution is on the impact of tissue of experimental diabetic mice whole blood LPO content
Diabetic groups experimental mouse LPO content after the modeling obviously raises than the LPO content of control group experimental mouse, gives behind the mulberry leaf extracting solution whole blood LPO content obviously descend (P＜0.05).The amount difference of giving of its Folium Mori extract also has this different effect.The results are shown in Table 3:
Table 3 mulberry leaf extracting solution group is on the impact of artificial diabetes experimental mouse whole blood LPO content
Table 3 The effects of extract of mulberry leaves on LPO in diabetic rats (Jian x ± s, mmol/L)
As shown in Table 3, Folium Mori extract has certain effect to the reduction of LPO in the blood glucose in diabetic mice, and along with the rising of concentration, its effect is also more obvious, and the LPO content of Folium Mori extract group 3 is also close to normal level.Glycosuria pathology group is then unchanged, and the LPO content of injection of insulin group mouse has then approached normal level.
Claims (1)
1. the preparation method of a polysaccharides of Folium Mori, it comprises the following steps:
Pulverize dry mulberry leaf 10kg, add 180 L distilled water, 70 ℃ of heating and refluxing extraction 90min, three layers of filtered through gauze twice are used ceramic membrane filter again, are evaporated to 10 L, then concentrated solution removes albumen with 10% trichoroacetic acid(TCA), upper SEPHADEX G-10 post separated and collected contains the polysaccharides of Folium Mori component, is concentrated into 1L and goes up silicagel column separated and collected polysaccharides of Folium Mori composition again and be concentrated into driedly, obtains polysaccharides of Folium Mori.
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| CN103127227A (en) * | 2011-11-28 | 2013-06-05 | 山东农业大学 | Preparation method for mulberry leaf polysaccharide hypoglycemic active component |
| CN112220825A (en) * | 2019-07-14 | 2021-01-15 | 北京恒润泰生医药科技有限公司 | Mulberry leaf aqueous extract powder, preparation, immune enhancement effect and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1850166A (en) * | 2006-03-08 | 2006-10-25 | 江苏大学 | Composition of white mulberry leaf blood-sugar-reducing effective components and preparing method |
| CN101648852A (en) * | 2009-09-02 | 2010-02-17 | 唐忠海 | Method for rapidly extracting resveratrol from mulberry leaves |
| CN101712726A (en) * | 2009-10-30 | 2010-05-26 | 浙江工业大学 | Method for preparing high-purity mulberry leaf polysaccharide |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1850166A (en) * | 2006-03-08 | 2006-10-25 | 江苏大学 | Composition of white mulberry leaf blood-sugar-reducing effective components and preparing method |
| CN101648852A (en) * | 2009-09-02 | 2010-02-17 | 唐忠海 | Method for rapidly extracting resveratrol from mulberry leaves |
| CN101712726A (en) * | 2009-10-30 | 2010-05-26 | 浙江工业大学 | Method for preparing high-purity mulberry leaf polysaccharide |
Non-Patent Citations (2)
| Title |
|---|
| 桑叶多糖的提取纯化及其含量测定;欧阳臻等;《时珍国医国药》;20051231;第16卷(第10期);961-962 * |
| 欧阳臻等.桑叶多糖的提取纯化及其含量测定.《时珍国医国药》.2005,第16卷(第10期),961-962. |
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