CN101597389B - Mixture of momordica saponins and polysaccharides and extraction process and application thereof - Google Patents
Mixture of momordica saponins and polysaccharides and extraction process and application thereof Download PDFInfo
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Abstract
The invention discloses a mixture of momordica saponins and polysaccharides, which is obtained by the following preparation methods: the balsam pear is used as a raw material and is added with an organic solvent to undergo ultrasound-microwave assisted extraction; a momordica saponins extract is obtained after filtration; momordica filter residues are extracted in alkaline solution; the obtained extract is namely the momordica polysaccharides extract; the momordica saponins extract and the momordica polysaccharides extract are respectively concentrated, dried and mixed to obtain a mixture of momordica polysaccharides and saponins; the invention also discloses an extraction process and application of the mixture of the momordica saponins and polysaccharides; the mixture has various physiological functions such as blood sugar level reduction, cancer resistance, virus resistance, blood lipids regulation, immunity increasing and the like; the mixture can be used for preparing health food with the function of reducing blood sugar level; the extraction process adopts step-by-step extraction technology and has the characteristics of high raw materials utilization rate, mild extraction conditions, high speed, high efficiency and the like; compared with the single extraction technology, for the extraction process of the invention, the extraction time is shortened by more than 20% and the purity of the product is improved by more than 15%.
Description
Technical field
The invention belongs to the processing of farm products field, be specifically related to a kind of momordica saponins and polysaccharide mixture and extraction process thereof and application.
Background technology
Balsam pear is famous Chinese medicine among the people and vegetables commonly used at the existing nearly 500 years applicating history of China.The successive dynasties clinical application is in diseases such as treatment tumour, dysentery, heatstrokes.Compendium of Material Medica record: balsam pear " bitter cold, nontoxic, remove heat symptoms caused by an exopathgen, separate tired, clear away heart-fire make eye bright, beneficial gas establishing-Yang ".Modern times studies show that in a large number, and balsam pear contains abundant polysaccharide and saponin class biologically active substance, have the different physiological roles of hypoglycemic, anticancer, antiviral, regulating blood fat, strengthening immunity etc.Therefore, the method for the simultaneous extraction of research bitter melon polysaccharide and saponin has important development and application values.
At present about bitter melon polysaccharide or the existing more report of the independent technology of extracting of saponin(e, extraction method to bitter melon polysaccharide is mainly taked traditional water formulation, zymohydrolysis extracting method, membrane separation process, and the extracting method of momordica saponins is mainly organic solvent extractionprocess, hot water extraction method, microwave extraction method etc.Based on bitter melon polysaccharide and the independent extraction method technology of saponin(e length consuming time, the temperature height causes that easily impurity is too much, problems such as active substance degraded, be necessary to find a raw material availability height, extraction conditions gentleness, unite extracting method fast and efficiently, and correlative study does not also appear in the newspapers.
Summary of the invention
The object of the present invention is to provide a kind of momordica saponins and polysaccharide mixture, that this momordica saponins and polysaccharide mixture have is hypoglycemic, anticancer, antiviral, the different physiological roles of regulating blood fat, strengthening immunity etc.
The present invention also aims to provide the extraction process of above-mentioned momordica saponins and polysaccharide mixture, this extraction process raw material availability height, extraction conditions gentleness, and improved momordica saponins and extracting efficiency of polysaccharides.
The present invention also aims to provide above-mentioned momordica saponins and polysaccharide mixture to have application in the protective foods of hypoglycemic activity in preparation.
For reaching first purpose of the present invention, momordica saponins provided by the invention and polysaccharide mixture, prepare by the following method: be raw material with the dried Frutus Momordicae Charantiae, add organic solvent, carry out ultrasonic-microwave cooperating and extract, get the momordica saponins extracting solution after the filtration, the balsam pear filter residue is carried out alkali lye to be extracted, the gained extracting solution is the bitter melon polysaccharide extracting solution, to momordica saponins extracting solution and bitter melon polysaccharide extracting solution concentrate, dry and mix after, promptly get bitter melon polysaccharide and saponin mixture.
For reaching second purpose of the present invention, the extraction process of above-mentioned momordica saponins provided by the invention and polysaccharide mixture, may further comprise the steps: get dried Frutus Momordicae Charantiae, after micronizing, add organic solvent, carrying out ultrasonic-microwave cooperating extracts, get the momordica saponins extracting solution after the filtration, the balsam pear filter residue is carried out alkali lye to be extracted, the gained extracting solution is the bitter melon polysaccharide extracting solution, respectively to momordica saponins extracting solution and bitter melon polysaccharide extracting solution concentrate, dry and mix after, can obtain bitter melon polysaccharide and saponin.
Further, the extraction process of above-mentioned momordica saponins provided by the invention and polysaccharide mixture may further comprise the steps:
(1) micronizing: get dried Frutus Momordicae Charantiae, micronizing is carried out in chopping, obtains the ultra micro Fructus Momordicae charantiae powder;
(2) ultrasonic-microwave cooperating extracts: the organic solvent that adds its 12～24 times of weight in the ultra micro Fructus Momordicae charantiae powder, the condition of regulating ultrasonic-microwave cooperating processing is: power 40～60W, frequency 15～25khz, carry out ultrasonic-microwave cooperating and extract 800～1150s, filter gained filtrate and be the momordica saponins extracting solution;
(3) alkali lye extracts: add the alkali lye of its 30～40 times of weight in above-mentioned filtration gained filter residue, regulating pH is 7～9, and temperature is 80～90 ℃, extracts 20～30min, filters gained solution and is the bitter melon polysaccharide extracting solution;
(4) concentrate and dry: to the momordica saponins extracting solution and the bitter melon polysaccharide extracting solution concentrates and drying after, can obtain the mixture of bitter melon polysaccharide and saponin by 1: 1 weight ratio mixing.
The granularity of Fructus Momordicae charantiae powder is 250～350 orders in the above-mentioned steps (1).
Volume of organic solvent concentration is 65～85% ethanol in the above-mentioned steps (2).
Alkali lye is aqueous sodium hydroxide solution in the above-mentioned steps (3).
The volume that concentrates back momordica saponins extracting solution and bitter melon polysaccharide extracting solution in the above-mentioned steps (4) is 1/10～1/5 of an original volume.
Momordica saponins that the present invention extracts and polysaccharide mixture have application in the protective foods of hypoglycemic activity in preparation.
The bitter melon polysaccharide of the inventive method preparation and saponin mixture can be prepared into bromatology the above formulation or make an addition in the protective foods in the mode of active substance toughener.
The invention has the beneficial effects as follows:
(1) momordica saponins of the present invention and polysaccharide mixture have the different physiological roles of hypoglycemic, anticancer, antiviral, regulating blood fat, strengthening immunity etc.;
(2) the present invention has adopted momordica saponins and polysaccharide and has extracted treatment technology step by step, can solve complex process in the independent extractive technique effectively, the extraction temperature is too high, shortcomings such as product quality is not good, have raw material availability height, extraction conditions gentleness, characteristics such as quick, efficient, compare with independent extractive technique, extraction process of the present invention shortened more than 20% on extraction time, and product purity improves more than 15%;
(3) the present invention has adopted ultrasonic-microwave cooperating to extract treatment technology to momordica saponins and polysaccharide, and extraction conditions such as the temperature extracted, time, pH, solid-liquid ratio, granularity, alcoholic strength are optimized, under this optimal condition, the extraction yield of balsam pear saponin(e can reach 2.51%, the extraction rate reached 12.86% of bitter melon polysaccharide exceeds more than 18% and 25% than independent extractive technique technology respectively;
(4) momordica saponins and the polysaccharide of the inventive method preparation, after 1: 1 weight ratio mixing, can make an addition in the mode of active substance toughener in the product such as protective foods, bread and cheese of diabetes, market potential is huge, this to development of promoting balsam pear deep processing industry, improve its economic benefit, the Sustainable development that promotes agricultural is significant.
Description of drawings
Fig. 1 is the mouse glucose tolerance curve;
Fig. 2 respectively organizes mouse islets tissue pathologies change, wherein Fig. 2 a is the blank group, 2b is a model group, 2c is balsam pear saponin(e (I), Fig. 2 d is balsam pear saponin(e (II), and Fig. 2 e is bitter melon polysaccharide (I), and Fig. 2 f is bitter melon polysaccharide (II), Fig. 2 g is bitter melon polysaccharide and saponin(e mixture (I), and Fig. 2 h is bitter melon polysaccharide and saponin(e mixture (II).
Embodiment
Following examples only are used to set forth the present invention, and protection scope of the present invention is not only to be confined to following examples.The those of ordinary skill of described technical field all can be realized purpose of the present invention according to above content disclosed by the invention and scope that each parameter is got.
First part's momordica saponins and polysaccharide mixture and extraction process thereof and application
Wherein, embodiment 1～3 is momordica saponins of the present invention and polysaccharide mixture and extraction process thereof; Embodiment 4～5 has application in the protective foods of hypoglycemic activity for momordica saponins of the present invention and polysaccharide mixture in preparation.
Embodiment 1
Get dried Frutus Momordicae Charantiae 1kg, micronizing to 250 order adds volumetric concentration and is 75% ethanolic soln 18kg, soaks 1h, and the ultrasonic-microwave cooperating at 60w, 20khz extracts 800s then; Said extracted liquid is filtered, and filtrate is the momordica saponins extracting solution; To above-mentioned filtration gained filter residue, add the feed liquid weight ratio and be 1: 35, pH and be 9 the NaOH aqueous solution and extract 25min down at 80 ℃, filter then, filter obtained aqueous solution and be the bitter melon polysaccharide extracting solution; Respectively above-mentioned gained momordica saponins extracting solution and bitter melon polysaccharide extracting solution are carried out concentration to 1/7.5 of original volume, obtain momordica saponins concentrated solution and bitter melon polysaccharide concentrated solution; Respectively momordica saponins concentrated solution and bitter melon polysaccharide concentrated solution are carried out lyophilize, can obtain bitter melon polysaccharide and saponin mixture after mixing by 1: 1 weight ratio.
Embodiment 2
Get dried Frutus Momordicae Charantiae 1kg, micronizing to 300 order adds volumetric concentration and is 65% ethanolic soln 12kg, soaks 1h, and the ultrasonic-microwave cooperating at 50w, 25khz extracts 1150s then; Said extracted liquid is filtered, and filtrate is the momordica saponins extracting solution; To above-mentioned filtration gained filter residue, add the feed liquid weight ratio and be 1: 40, pH and be 8 the NaOH aqueous solution and extract 20min down at 85 ℃, filter then, filter obtained aqueous solution and be the bitter melon polysaccharide extracting solution; Respectively above-mentioned gained momordica saponins extracting solution and bitter melon polysaccharide extracting solution are carried out concentration to 1/5 of original volume, obtain momordica saponins concentrated solution and bitter melon polysaccharide concentrated solution; Respectively momordica saponins concentrated solution and bitter melon polysaccharide concentrated solution are carried out lyophilize, can obtain bitter melon polysaccharide and saponin mixture after mixing by 1: 1 weight ratio.
Embodiment 3
Get dried Frutus Momordicae Charantiae 1kg, micronizing to 350 order adds volume integral concentration and is 85% ethanolic soln 24kg, soaks 1h, and the ultrasonic-microwave cooperating at 40w, 15khz extracts 975s then; Said extracted liquid is filtered, and filtrate is the momordica saponins extracting solution; To above-mentioned filtration gained filter residue, add the feed liquid weight ratio and be 1: 30, pH and be 7 NaOH solution and extract 30min down at 90 ℃, filter then, filter obtained aqueous solution and be the bitter melon polysaccharide extracting solution; Respectively above-mentioned gained momordica saponins extracting solution and bitter melon polysaccharide extracting solution are carried out concentration to 1/10 of original volume, obtain momordica saponins concentrated solution and bitter melon polysaccharide concentrated solution; Respectively momordica saponins concentrated solution and bitter melon polysaccharide concentrated solution are carried out lyophilize, can obtain bitter melon polysaccharide and saponin mixture after mixing by 1: 1 weight ratio.
Embodiment 4
Present embodiment momordica saponins and polysaccharide mixture and tea etc. share, weight proportion is between 1: 1～10, electuary, every day, 10～100g can be used for the prevention, strengthening immunity of the diabetes of diabetics's hypoglycemic activity and healthy population, antitumor and effect such as delay senility.
Present embodiment pulverulent water-soluble active substance momordica saponins and polysaccharide mixture make an addition to protective foods in the mode of active substance toughener, weight proportion is 1: 1～5, can be used for the prevention, strengthening immunity of the diabetes of diabetics's hypoglycemic activity and healthy population, antitumor and effect such as delay senility.
(2) test of pesticide effectiveness of momordica saponins and polysaccharide mixture
1, test method
1.1 sample
The momordica saponins and the bitter melon polysaccharide mixture of the present invention's preparation.
1.2 main agents
Streptozotocin (STZ) AR U.S. Sigma company; The glucose assays test kit is available from Shanghai Rongsheng Bioisystech Co., Ltd; Superoxide-dismutase (SOD) testing cassete, catalase (CAT) testing cassete, testing cassete mda (MDA) is measured test kit, gsh (GSH) (removing albumen) is measured test kit, hexokinase (HK) testing cassete, the Xylene Brilliant Cyanine G protein determination kit all builds up bio-engineering research institute available from Nanjing.
1.3 instrument
CO
2Incubator, enzyme-linked immunosorbent assay instrument, ultraviolet-visible pectrophotometer, analytical balance, vacuum freeze drier, Rotary Evaporators.
1.4 laboratory animal
Kunming kind small white mouse (20 ± 2g), the SPF level, male, Zhongshan University's Experimental Animal Center provides, and kunming mice is divided into 8 groups, 12/group, raises under the condition of 22 ℃ of temperature and relative humidity 60% in Animal House.
1.5 animal is handled and the preparation of tissue homogenate
Get hepatic tissue blocking (about 0.5g), in pre-ice-cold physiological saline rinsing clean, wipe away dried with filter paper.Put into the centrifuge tube of ice bath after weighing, the physiological saline that adds 9 times of volumes prepares tissue homogenate with tissue refiner.10% liver homogenate that will prepare the again centrifugal 10～15min of refrigerated centrifuge (4 ℃) 3000rpm, it is to be measured to get supernatant liquor.
The protein content of supernatant liquor is measured with the Coomassie brilliant blue method.
1.6DM the foundation of mouse model
Adaptability is fed mouse fasting 15h behind the 7d, STZ130mg/ (kgbw) dosage abdominal injection, STZ 0.1mol/L, the fresh preparation of the sodium citrate buffer solution of pH=4.2 (operation on ice), fasting plasma glucose (BG) is surveyed in the blood sampling of 3d posterior orbit venous plexus, and BG 〉=11.0mmol/L person is the DM mouse.
1.7 laboratory animal grouping and blood sugar detection thereof
The DM mouse is divided into 7 groups at random, and promptly model group and 6 experimental group are established the normal control group simultaneously; The blank group is irritated stomach physiological saline, and experimental group is irritated stomach respectively and given 150,300mg (the balsam pear saponin(e group of (kgbw), bitter melon polysaccharide group, and the two mixture group, specific design such as following table:
Grouping of table 1 animal and disposition
Experiment periods is 30d, and regularly administration every day was got blood once in 10 days at interval, detected fasting plasma glucose, and before the detection, mouse is changed bedding and padding and fasting 12h, presses glucose oxidase enzyme reagent kit specification sheets and measures blood glucose value.
1.8 glycolipid metabolism
1.8.1 the mensuration of mouse sugar tolerance
The DM mouse is being measured its sugar tolerance behind the treatment 30d continuously, measure blood sugar concentration (0h) the last time after the administration immediately, and give normal control group, model control group, saponin(e low dose group, saponin(e high dose group, polysaccharide low dose group, polysaccharide high dose group, mutual low dose group and mutual high dose group, dosage is 1g/kgbw, measures 30,60 respectively after administration, the blood sugar concentration of 120min mouse.
1.8.2 mouse lipid determination
Behind the 30d, put to death mouse, separation of serum is pressed the test kit operational requirement and is surveyed three of blood fat: TC, TG and HDL-C.
1.8.3 the mensuration of liver starch
Measuring principle: glycogen can dewater under the effect of the vitriol oil and generate the alditol derivative, and the latter forms blue compound with the anthrone effect again, with the standard glucose solution colorimetric assay of handling with method; Glycogen is highly stable in concentrated alkali solution, keeps glycogen so earlier tissue was put into the concentrated base heating before colour developing to destroy other composition.
1.8.4 the mensuration of liver hexokinase
Measuring principle: use the linked reaction of glucose-6-phosphate dehydrogenase (G-6-PD),,, reflect the activity of HK by the increase of measuring absorbancy at 340nm wavelength place providing under the substrate condition of capacity.
Unit definition: at 37 ℃, under the condition of pH7.6, the NADPH that every gram tissue protein per minute in this reaction system generates 1mmol/L is defined as an enzyme activity unit.
1.9 antioxygenation
1.9.1 the mensuration of superoxide-dismutase (SOD) vigor
Measuring principle: produce ultra-oxygen anion free radical (O by xanthine and XOD reactive system
2 -), latter's oxidation azanol forms nitrite, presents red-purple under the effect of developer, surveys its absorbancy with visible spectrophotometer.When containing SOD in the sample, then ultra-oxygen anion free radical there is narrow spectrum restraining effect, the nitrite of formation is reduced, the light absorption value of measuring pipe during colorimetric is lower than the light absorption value of control tube, by calculating the SOD vigor that can obtain sample.
SOD vigor definition in the tissue homogenate: it is a SOD unit of activity (U) that every milligram of tissue protein SOD inhibiting rate in the 1mL reaction solution reaches 50% o'clock pairing SOD.
1.9.2 mda (MDA) Determination on content
Measuring principle: the mda in the lipid peroxide degraded product (MDA) can with thiobarbituricacid (TBA) condensation, form red product, this product has maximum absorption band at the 532nm place.
1.9.3 catalase (CAT) vitality test
The reaction of catalase decomposing H 2O2 can be ended rapidly by adding ammonium molybdate, and remaining H2O2 and ammonium molybdate effect produce a kind of flaxen complex compound, measures its growing amount at the 405nm place, can calculate the vigor of CAT.
Definition: every milligram of tissue protein decomposes the H of 1 μ mol p.s.
2O
2Amount be a unit of activity.
1.9.4 the mensuration of gsh (GSH)
Can produce a kind of yellow compound when dithio dinitrobenzoic acid and sulfhydryl compound reaction, can carry out colorimetric assay and measure.
1.10 pancreas islet provide protection
1.10.1C-the mensuration of peptide
Measuring principle: adopt the competition radio immunoassay, promptly C-P in standard or the testing sample and 125I-C-P common with the specific C-P antibody of limiting the quantity of being at war with property association reaction under suitable condition.
All test by the test kit explanation
1.10.2 pancreatic tissue section HE dyeing is observed
1. draw materials
After mouse cervical vertebra dislocation is put to death, get pancreas rapidly and place and contain 4% neutral formalin stationary liquid (pH7.2), fix 24 hours, conventional dehydration embedding.
2. get wax stone and cut into slices, thickness 5 μ m, conventional H E dyeing, om observation after the mounting.
1.11 mouse small intestine mucous membrane alpha-glucosidase activity relatively
1.11.1 mucous membrane of small intestine homogenate preparation method
Mouse is taken from duodenum and begins to jejunum mucous membrane of small intestine 10cm after putting to death, and takes out small intestine immediately and places on the ice platform, cuts small intestine open and exposes intestinal mucosa, and is dried with wiping away behind the ice-cold normal saline flushing, scrapes with slide and gets mucous membrane of small intestine.Scrape and get the gained mucous membrane of small intestine and weigh, add ice-cold PBS by a certain percentage, homogenate in ice bath, mucous membrane homogenate is got supernatant as the alpha-glucosidase suspension in the centrifugal 15min of low temperature (4 ℃) 3000rpm.Press the bright blue protein determination kit specification sheets operation of Kao Masi, measure total protein content in each mucous membrane of small intestine homogenate.According to total protein content,, make per 50 μ L mucous membrane of small intestine homogenate contain about 100 μ g total proteins with physiological saline dilution preparation alpha-glucosidase test suspension.
1.11.2 the mucous membrane of small intestine alpha-glucosidase activity is measured
The method of reference literature adds maltose (or sucrose, the lactose) solution of 42mmol/L in the test tube, (66.7mmol/L pH5.6), is hatched 20min for 37 ℃ to PBS, adds 0.2mol/LNa
2CO
3Termination reaction.
The centrifugal 15min of reaction solution 3000rpm gets supernatant.Press the operation of glucose assays test kit specification sheets,
Mensuration supernatant liquor glucose content (Gs, mmol/L).Calculate the glucose amount (Gp, mmol/g protein) that the catalysis of the every g protein of mucous membrane of small intestine homogenate generates:
Gp(mmol/g?protein)＝(Gs·0.225)x10
1.11.3 mouse small intestine mucous membrane alpha-glucosidase activity relatively
Generate glucose yield (mmol/gprotein) according to every gram albumen catalysis lactose, sucrose and maltose in the mucous membrane of small intestine.
1.12 data analysis
The data SPSS 10.0 softwares are handled, and carry out relatively reaching between the multiple sample mean variance analysis of comparing in twos; If heterogeneity of variance is then compiled the variance analysis of comparing in twos again after the order.There is statistical significance P＜0.05 for difference.
2, test-results
2.1 influence to DM mouse blood sugar value due to the STZ
Find out by following table 2, by being observed, 30 days change of blood sugar of DM mouse find, the administration group all can significantly suppress the blood sugar increasing of DM mouse, and administration after 21 days except that saponin(e low dose group and polysaccharide low dose group blood sugar decreasing effect significantly (P＞0.05), all the other each organize and all significantly be better than model group (P＜0.05).When experiment finished, each administration group blood glucose value all significantly was better than model group (P＜0.05), and there are significant dose-effect relationship (P＜0.05) in saponin(e, polysaccharide and mutual group.Saponin(e high dose group mouse blood sugar value descends 9.8% than saponin(e low dose group mouse blood sugar value, polysaccharide high dose group mouse blood sugar value descends 8.5% than polysaccharide high dose group mouse blood sugar value, and the mutual low dose group mouse blood sugar value of mutual high dose group mouse blood sugar value descends 8.8%.And mutual low dose group mouse blood sugar value significantly is lower than saponin(e low dose group and polysaccharide low dose group (P＜0.05), reaches 8.9% and 11.0% respectively; Mutual high dose group mouse blood sugar value significantly is lower than saponin(e high dose group and polysaccharide high dose group (P＜0.05), reaches 8.0% and 11.3% respectively.As can be seen, bitter melon polysaccharide and saponin(e are induced and are existed synergistic function aspect the hyperglycemia mouse blood sugar value reducing STZ.
Table 2 momordica saponins and polysaccharide and composition thereof to the influence of DM mouse blood sugar value due to the STZ (x ± S, n=12)
2.2 to DM mouse sugar tolerance influence due to the STZ
The sugar tolerance curve of mouse is seen accompanying drawing 1, exogenous glucose can cause blood sugar increasing, the glucose tolerance experiment is one of test of making a definite diagnosis DM, under the normal circumstances, because the coordinative role of mechanism of blood glucose regulation, even once take in a large amount of sugar, blood sugar concentration also only can temporarily raise, soon promptly recover normal level, this phenomenon is called anti-sugared phenomenon, generally observe anti-sugared phenomenon with carbohydrate tolerance test, carbohydrate tolerance test is divided into oral glucose test (OGTT), test glucose tolerance by intravenous infusion, methods such as cortisone glucose tolerance test, wherein the former is the most commonly used, can understand individual portative power to glucose by glucose tolerance test, the functional status of assessment islet cells.
From following table 3 as can be known, normal control group mouse sugar tolerance is normal, take glucose after, its blood glucose value reaches highest level when 30min, descend gradually subsequently, drops to normal level during 120mim; Its blood sugar of model control group reaches highest level when 30min, maintain higher level thereafter always, shows as the feature that sugar tolerance reduces.
Each administration group all can suppress the rising at hyperglycemia mouse blood sugar peak, and the blood sugar fall almost drops to the preceding level of administration far above model control group during 120min; Polysaccharide shows significant dose-effect relationship to the DM mouse, and polysaccharide high dose group mouse area under curve significantly is lower than the polysaccharide low dose group and reaches 6.3%; And mutual high dose group is optimum in each administration group, significantly reduce area under curve (P＜0.05) than saponin(e low dosage, saponin(e high dosage, polysaccharide low dosage, polysaccharide high dosage and mutual low dose group mouse, reach 6.5,3.8,10.0,3.9 and 4.1% respectively.
Table 3 momordica saponins and polysaccharide and composition thereof to DM mouse sugar tolerance influence due to the STZ (x ± S, n=12)
2.3 to liver starch, hexokinase in the DM mouse liver due to the STZ
The basic source of blood sugar is the carbohydrate in the food, and when blood sugar was tending towards reducing not taking food, the liver starch Decomposition was strengthened, and when long-term hunger, the liver glyconeogenesis strengthens, thereby blood sugar still can continue to maintain normal level.
To be glucosyl residue with α-(1,4) glycosidic link link to each other glycogen constitutes straight chain and constitute many ramose macromolecular polymers that contain of side chain with α-(1,6) glycosidic link, mainly is stored in muscle and the liver.Cross when low when glucose concn in the body, the muscle self that is decomposed into of muscle glycogen shrinks energize, and the decomposition of liver starch then is used for keeping blood sugar concentration, for neural system and other histoorgans provide every day required energy.If blood sugar concentration is too high in the body, too high glucose concn can be induced β emiocytosis Regular Insulin again, is that liver starch is stored with conversion of glucose, with lowering blood glucose.
Irritating stomach, respectively to organize in the mouse liver glycogen content after 30 days as shown in table 4 below, and hepatic glycogen content is remarkable decline (P＜0.01) in the DM model group mouse liver, has only 36% of blank group Mouse Liver glycogen content.Influence all is significant dose-effect relationship (P＜0.05) to the Mouse Liver glycogen content for saponin(e, polysaccharide and mutual group, the saponin(e high dose group rises 26.2% than the saponin(e low dose group, the polysaccharide high dose group rises 57% than the polysaccharide low dose group, and the mutual low dose group of mutual high dose group rises 21.4%.And low dose group Mouse Liver glycogen content is significantly higher than saponin(e low dose group and polysaccharide low dose group (P＜0.05) alternately, reaches 20.8%, 86.3% respectively; Mutual high dose group Mouse Liver glycogen content also is significantly higher than saponin(e high dose group and polysaccharide low dose group (P＜0.05), reaches 13.5%, 44.1% respectively, and mutual group is remarkable synergistic function.
The first step of glucose metabolism is the phosphorylation of glucose, and at first insulin stimulating glucose transfer protein 4 (GLUT4) is transferred to glucose in the Skeletal Muscle Cell, and hexokinase (HK) catalysis conversion of glucose is 6-glucose 1-phosphate1-(G-6-P) then.HK comprises 4 kinds of isomerases: HK I-IV, wherein IV type HK (glucokinase) is present in the liver cell, plays an important role in the glucose phosphorylation process of insulin-mediated.
As shown in table 4 below, irritate stomach after 30 days, the HK enzyme is lived and is extremely significantly descended (P＜0.01) in the DM model group mouse liver, has only 30% of blank group; And each administration group all can significantly improve HK enzyme (P＜0.05) alive in the DM mouse liver; And saponin(e and polysaccharide there is remarkable dose relationship (P＜0.05) in the influence of living to the HK enzyme, the saponin(e high dose group is high by 21.0% than the saponin(e low dose group, the polysaccharide high dose group is high by 51.4% the polysaccharide low dose group; The mutual no remarkable dosage influence of group (P＞0.05), but mutual low dose group reaches 23.7% and 97.9% respectively than significantly increase (P＜0.05) alive of HK enzyme in saponin(e low dose group and the polysaccharide low dose group mouse liver.
Table 4 momordica saponins and polysaccharide and composition thereof are to the influence of liver starch, hexokinase in the DM mouse liver due to the STZ (x ± S)
2.4 influence to DM mouse blood lipid level due to the STZ
It is unusual that abnormal carbohydrate metabolism often merges lipid metabolism.A large amount of blood fat is deposited on the intra-arterial tube wall, in blood vessel, form atheromatous plaque, patch increases gradually and increases, the hardening of vessel wall progressive additive, lumen of vessels diminishes, and causes the position ischemic of this vascularity in the course of time, blood supply is interrupted when serious, this pathology can occur in the whole body blood vessel, and leads to complications, and is the early dead and major cause of morbidity of patient DM.
The filling stomach is respectively organized the mouse Blood Lipid after 30 days as shown in table 5 below, saponin(e, polysaccharide group mouse total cholesterol (TC) level are significant dose-effect relationship, high dosage all significantly is better than low dose group (P＜0.05) and reaches 14.3,13.0% respectively, though the TC level does not have significant difference (P＞0.05) between the mutual low high dose group mouse, but mutual high dose group mouse significantly is lower than polysaccharide high dosage and saponin(e high dosage mouse, reaches 3.2,23.0% respectively.
Low density lipoprotein cholesterol (LDL-C) and triglyceride level (TG) level also all are significantly higher than blank group mouse (P＜0.05) in the DM model group mice serum, each administration group all can significantly reduce LDL-C and TG level, but reduces horizontal effect of TG and indifference between each administration group.Saponin(e, polysaccharide group mouse low density lipoprotein cholesterol (LDL-C) level are significant dose-effect relationship, high dosage all significantly is better than low dose group (P＜0.05) and reaches 11.7,12.6% respectively, though the LDL-C level does not have significant difference (P＞0.05) between the mutual low high dose group mouse, but significantly be lower than polysaccharide high dosage and saponin(e high dosage mouse, reach 4.3,18.2% respectively.
The effective hypoglycemic component of table 5 balsam pear to the influence of DM mouse blood fat due to the STZ (x ± S, n=12)
2.5 influence to oxidative stress level in the DM mouse liver due to the STZ
By following table 6 as can be known, irritate stomach and respectively organize catalase in the mouse liver (CAT), superoxide-dismutase (SOD), mda (MDA) and reductive glutathione (GSH) level difference after 30 days.Balsam pear is respectively organized extract and is irritated stomach after 30 days, mutual high dose group (300mg/kgbw) murine liver tissue antioxidant level all is better than or significantly is better than all the other DM mouse (P＜0.05), improve CAT level (reaching 11.1,6.7,24.8,4.3 and 16.0% respectively), SOD level (reaching 30.1,14.7,15.6,3.7 and 17.7% respectively) than saponin(e low dosage, saponin(e high dosage, polysaccharide low dosage, mutual low dose group mouse, reduce MDA level (reaching 26.1,16.8,12.2,3.0 and 8.2% respectively).
The GSH level also significantly is lower than blank group mouse in the DM mouse liver, saponin(e group and mutual group mouse GSH water-glass reveal significant dose relationship (P＜0.05) as can be seen by following table 6, the saponin(e high dose group improves 11.4% than saponin(e low dose group mouse liver GSH level, and mutual high dose group improves 14.0% than saponin(e low dose group mouse liver GSH level.The GSH level significantly is better than saponin(e low dose group, saponin(e high dose group, polysaccharide low dose group and polysaccharide high dose group (P＜0.05) in the mutual high dose group DM mouse liver, reaches 22.9,10.3,25.2 and 14.0 respectively.
The effective hypoglycemic component of table 6 balsam pear is to CAT in the DM mouse liver due to the STZ, SOD, and MDA, the influence of GSH (x ± S, n=12)
2.6 restraining effect to DM mouse small intestine alpha-glycosidase due to the STZ
By following table 7 as can be known, irritate stomach and respectively organize mouse small intestine mucous membrane sucrase, Sumylact L and maltase activity difference after 30 days.Every gram mucous membrane of small intestine homogenate catalysis sucrose, maltose and the glucogenic amount of lactose are all than blank group significantly rise (P＜0.05) in the DM model group 20 minutes.Saponin(e and mutual group suppress to be remarkable influence (P＜0.05) to sucrase, mutual high dose group mouse small intestine sucrase and maltin enzyme activity level significantly are lower than saponin(e low dosage, saponin(e high dosage, polysaccharide low dosage, mutual low dose group mouse (P＜0.05), wherein reduce mouse small intestine sucrase vigor and reach 35.8,20.2,27.5,24.2 and 31.2% respectively, reduce mouse small intestine maltin enzyme activity and reach 8.6,6.5,16.9,13.6 and 16.5% respectively.
Aspect inhibition Sumylact L vigor, have only saponin(e to present significant dose relationship (P＜0.05) in each group, saponin(e low dose group mouse small intestine Sumylact L vigor is than saponin(e high dose group low 21.0%.Though mutual high dose group mouse small intestine Sumylact L vigor significantly is lower than polysaccharide low dose group and polysaccharide high dose group (P＜0.05), reaches 29.6,17.4% respectively, is significantly higher than saponin(e low dose group (P＜0.05) and reaches 26.7%.
The effective hypoglycemic component of table 7 balsam pear to DM mouse small intestine sucrase Sumylact L due to the STZ and maltin restraining effect (x ± S, n=12)
2.7 to DM mouse due to the STZ to the mouse islets provide protection
As can be seen from Table 8, compare with blank group mouse, C-peptide level significantly descend (P＜0.05) in the model control group mice serum, saponin(e, polysaccharide and organize alternately that C-peptide level is significant dose relationship (P＜0.05) in the mice serum, saponin(e high dose group mouse C-peptide level is higher than the saponin(e low dose group and reaches 32.5%, polysaccharide high dose group mouse C-peptide level is higher than the polysaccharide low dose group and reaches 47.2%, and mutual high dose group mouse C-peptide level is higher than mutual low dose group and reaches 31.8%.C-peptide level significantly is better than (P＜0.05) or is better than the saponin(e low dose group and the polysaccharide low dose group in the mutual low dose group mice serum, reaches 6.0% and 66.0% respectively; C-peptide level significantly is better than (P＜0.05) or is better than the saponin(e high dose group and the polysaccharide high dose group in the mutual high dose group mice serum, reaches 5.4% and 48.7% respectively.
The effective hypoglycemic component of table 8 balsam pear is to the influence of DM mice serum C-peptide due to the STZ (x ± S)
By accompanying drawing 2 as can be seen, the normal rats pancreas islet is painted light shallow cell mass, clear border, form rule, pancreas islet inner cell marshalling, big or small consistent being evenly distributed.Nucleus is hyacinthine, justifies greatly, and it is clear to examine, and kernel is obvious, the abundant pale pink that is of endochylema.Become the pancreas islet of mould mouse to diminish, islet cells quantity reduces, even loses the pancreas islet structure, and atrophy fully, disintegration mix with surrounding tissue, is difficult to identification.Polysaccharide low dose group mouse islets shape is also irregular, and proliferation of fibrous tissue, glass become, and pancreas islet is as seen based on the cell infiltration of lymphocyte, monocyte; Beta Cell of islet vacuolar degeneration, necrosis, disappearance make pancreas islet be hollow state.And polysaccharide high dose group mouse islets area slightly increases, but the pancreatic islet endocrine structure disturbance, form is irregular, most karyon distortion, and pyknosis is differed in size, and vacuolar degeneration occurs.Low high dose group of saponin(e and the mutual accidental mild fibrosis of low dose group mouse islets, mutual high dose group mouse endocrine cell form is near normal group, and the border is clearer, and cell is arranged more neat.
Claims (6)
1. momordica saponins and polysaccharide mixture, it is characterized in that, prepare by the following method: be raw material with the dried Frutus Momordicae Charantiae, add organic solvent, carrying out ultrasonic-microwave cooperating extracts, get the momordica saponins extracting solution after the filtration, the balsam pear filter residue is carried out alkali lye to be extracted, the gained extracting solution is the bitter melon polysaccharide extracting solution, respectively momordica saponins extracting solution and bitter melon polysaccharide extracting solution are concentrated, after drying and the mixing, promptly get bitter melon polysaccharide and saponin mixture, described organic solvent is that volumetric concentration is 65～85% ethanol, and the condition that ultrasonic-microwave cooperating extracts is: power 40～60W, frequency 15～25khz, carry out ultrasonic-microwave cooperating and extract 800～1150s, described alkali lye is aqueous sodium hydroxide solution, and described momordica saponins and bitter melon polysaccharide mix by 1: 1 weight ratio.
2. the extraction process of described momordica saponins of claim 1 and polysaccharide mixture, it is characterized in that, may further comprise the steps: get dried Frutus Momordicae Charantiae, after micronizing, add organic solvent, carrying out ultrasonic-microwave cooperating extracts, get the momordica saponins extracting solution after the filtration, the balsam pear filter residue is carried out alkali lye to be extracted, the gained extracting solution is the bitter melon polysaccharide extracting solution, respectively momordica saponins extracting solution and bitter melon polysaccharide extracting solution is concentrated, dry and mix after, get final product bitter melon polysaccharide and saponin mixture, described organic solvent is that volumetric concentration is 65～85% ethanol, the condition that ultrasonic-microwave cooperating extracts is: power 40～60W, and frequency 15～25khz carries out ultrasonic-microwave cooperating and extracts 800～1150s, described alkali lye is aqueous sodium hydroxide solution, and described momordica saponins and bitter melon polysaccharide mix by 1: 1 weight ratio.
3. the extraction process of momordica saponins according to claim 2 and polysaccharide mixture is characterized in that, may further comprise the steps:
(1) micronizing: get dried Frutus Momordicae Charantiae, micronizing is carried out in chopping, obtains the ultra micro Fructus Momordicae charantiae powder;
(2) ultrasonic-microwave cooperating extracts: add the organic solvent of its 12～24 times of weight in the ultra micro Fructus Momordicae charantiae powder, carry out ultrasonic-microwave cooperating and extract, filter gained filtrate and be the momordica saponins extracting solution;
(3) alkali lye extracts: add the alkali lye of its 30～40 times of weight in above-mentioned filtration gained filter residue, regulating pH is 7～9, and temperature is 80～90 ℃, extracts 20～30min, filters gained solution and is the bitter melon polysaccharide extracting solution;
(4) concentrate and dry: respectively to the momordica saponins extracting solution with the bitter melon polysaccharide extracting solution concentrates and dry mixing afterwards can obtain bitter melon polysaccharide and saponin mixture.
4. the extraction process of momordica saponins according to claim 3 and polysaccharide mixture is characterized in that, the granularity of Fructus Momordicae charantiae powder is 250～350 orders in the step (1).
5. the extraction process of momordica saponins according to claim 3 and polysaccharide mixture is characterized in that, the volume that concentrates back momordica saponins extracting solution and bitter melon polysaccharide extracting solution in the step (4) is 1/10～1/5 of an original volume.
6. described momordica saponins of claim 1 and polysaccharide mixture have application in the protective foods of hypoglycemic activity in preparation.
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| CN101953875A (en) * | 2010-09-16 | 2011-01-26 | 天津农学院 | Bitter gourd extract composition for synergistically reducing blood sugar and preventing hyperglycemia |
| BR112013019251A2 (en) | 2011-01-28 | 2016-10-11 | Piramal Entpr Ltd | process for preparing an herbal extract |
| CN102772329B (en) * | 2012-07-11 | 2014-04-16 | 上海应用技术学院 | Bitter melon extract and preparation method thereof and application of bitter melon extract as moisturizer |
| RU2510278C1 (en) * | 2012-08-08 | 2014-03-27 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Тверской государственный университет" | Method for preparing saponin-containing extracts (versions) |
| CN106174696A (en) * | 2016-08-17 | 2016-12-07 | 福建中烟工业有限责任公司 | Prepare the method for Fructus Momordicae charantiae extract, Fructus Momordicae charantiae extract and application thereof |
| CN106490531A (en) * | 2016-10-21 | 2017-03-15 | 桂林淮安天然保健品开发有限公司 | The preparation method of balsam pear health-care product |
| CN108586560B (en) * | 2018-03-23 | 2020-07-17 | 汕头大学 | A two-aqueous-phase one-step extraction method of momordicoside and polysaccharide with blood glucose lowering activity |
| CN109430854B (en) * | 2018-09-18 | 2021-10-26 | 广东精核生物科技发展有限公司 | Health product and preparation method thereof |
| CN111067094B (en) * | 2019-11-28 | 2022-08-30 | 广东省农业科学院蚕业与农产品加工研究所 | Non-bitter gourd triterpene composition for inhibiting gluconeogenesis and application thereof |
| CN111053110A (en) * | 2019-12-27 | 2020-04-24 | 育生（厦门）生物科技有限公司 | Bitter gourd processing method and bitter gourd |
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Effective date of registration: 20160222 Address after: 510610 Tianhe District, Guangzhou, Dongguan, Zhuang Zhuang Road, No. 1, No. 133, Patentee after: Sericulture & Agri-Food Research Institute, GAAS Patentee after: Guangdong Bosun Health Food Co., Ltd. Address before: 510610 Tianhe District, Guangzhou, Dongguan, Zhuang Zhuang Road, No. 1, No. 133, Patentee before: Research Institute of Agricultural Biological Technology of GAAS Patentee before: Sericulture & Agri-Food Research Institute, GAAS Patentee before: Guangdong Bosun Health Food R & D Center |