CN105440150A - Carboxymethyl longan pulp polysaccharide and pharmaceutical application thereof - Google Patents
Carboxymethyl longan pulp polysaccharide and pharmaceutical application thereof Download PDFInfo
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Abstract
The invention relates to a carboxymethyl longan pulp polysaccharide. The carboxymethyl longan pulp polysaccharide is obtained by adding chloroacetic acid into a sodium hydroxide solution to react. The invention further relates to pharmaceutical application of the carboxymethyl longan pulp polysaccharide, in particular to application of the carboxymethyl longan pulp polysaccharide in preparation of anti-oxidation or immunological regulation medicaments. The application includes application of the carboxymethyl longan pulp polysaccharide in the preparation of free radial removing medicaments, lipid peroxidation restraining medicaments, red blood cell hemolysis restraining medicaments, T cell immune level raising medicaments, organism humoral immunity level raising medicaments and non-specific immunity enhancing medicaments. A synthetic carboxymethyl longan pulp polysaccharide is applied to in-vitro anti-oxidation and in-vivo immunological competence activity experiments of Kunming mice to obtain an experiment result about the anti-oxidation function and immunological regulation function of the carboxymethyl longan pulp polysaccharide, thereby laying a reference basis for the application of the carboxymethyl longan pulp polysaccharide in the medicine field.
Description
Technical field
The present invention relates to a kind of carboxymethylation Arillus longan polysaccharide and the application in pharmacy thereof.
Background technology
Molecular modification uses the methods such as physics, chemistry, biology to carry out structure of modification to compound molecule, to obtain the method for different types of structure derivative.Molecular modification is carried out to polysaccharide, the relative molecular mass of polysaccharide, viscosity, water solubility, charge number, space conformation etc. can be made to change, the biological activity strengthening natural polysaccharide even makes polysaccharide produce new activity, also can reduce the toxic side effect of polyose medicament, thus be conducive to the development & application of polyose medicament.It is introduce carboxymethyl on polysaccharide chain that carboxymethylation is modified, and carboxymethylated polysaccharides can improve solvability and the electronegativity of polysaccharide, and improves a lot to its anti-tumor activity, and can strengthen and even produce new biological activity.Such as, Auricularia polycose can make its anti-oxidant activity improve one times nearly after suitable carboxymethylation is modified; Carry out carboxymethylation modification to brave milk polysaccharide, obtain a kind of water-soluble good carboxymethylation tiger milk polysaccharide, it effectively can suppress Fe
2+the rat liver mitochondria lipid peroxidation that-Vc causes, the reduction of membrane fluidity and mitochondrial swelling; U. pertusa polysaccharide is after carboxymethylation is modified, and its reducing blood lipid been significantly enhanced; After astragalus polysaccharides is carried out carboxymethyl-modification, its growth promotion and enhancing immunocompetence are all clearly.Therefore, being necessary very much to carry out carboxymethylation modification to Arillus longan polysaccharide, for exploring structural modification to the bioactive impact of Arillus longan polysaccharide, being improved by molecular modification technology and the biological effect of developing Arillus longan polysaccharide lays the foundation.
Summary of the invention
The object of this invention is to provide the synthetic method of carboxymethylation Arillus longan polysaccharide, and obtain the result of immunity moderation effect in carboxymethylation Arillus longan polysaccharide antioxygenation and body.
The present invention achieves the above object by the following technical programs:
A kind of carboxymethylation Arillus longan polysaccharide, synthesized by following methods: Arillus longan polysaccharide is dissolved in sodium hydroxide solution, stirring reaction while adding Monochloro Acetic Acid, then reaction solution carried out dialysis is concentrated, obtain carboxymethylation Arillus longan polysaccharide after alcohol precipitation and lyophilize.
The synthetic method of described carboxymethylation Arillus longan polysaccharide is specially: be dissolved in by Arillus longan polysaccharide in 3mol/L sodium hydroxide solution, slowly add Monochloro Acetic Acid under vigorous stirring, makes its final concentration be 1.2mol/L; After continuing stirring reaction 3.2h after mixing at 73 DEG C, cooling, by HCl solution adjust ph to neutral; Reaction solution is placed in molecular weight cut-off be 3500 dialysis tubing to dialyse 48h, dialyzate, through concentrated, obtains carboxymethylation Arillus longan polysaccharide after alcohol precipitation and lyophilize.
The present invention is the claimed described application of carboxymethylation Arillus longan polysaccharide in pharmacy also.
Further, carboxymethylation Arillus longan polysaccharide comprises in following 7 preparing the application in medicine:
(1) carboxymethylation Arillus longan polysaccharide is preparing the application in anti-oxidant or immunoregulation druge.
(2) carboxymethylation Arillus longan polysaccharide is preparing the application in scavenging free radicals medicine, and carboxymethylation Arillus longan polysaccharide effective concentration is 2 ~ 12mg/ml.
(3) carboxymethylation Arillus longan polysaccharide is preparing the application of anti peroxidation of lipid medicine, and carboxymethylation Arillus longan polysaccharide effective concentration is 0.25 ~ 4.00g/L.
(4) carboxymethylation Arillus longan polysaccharide suppresses the application of erythrocyte hemolysis medicine in preparation, and carboxymethylation Arillus longan polysaccharide effective concentration is 28.60 ~ 457.1 μ g/ml.
(5) carboxymethylation Arillus longan polysaccharide improves the application in the horizontal medicine of T lymphocyte immunity in preparation, and carboxymethylation Arillus longan polysaccharide effective concentration is 20 ~ 160mg/kg.
(6) carboxymethylation Arillus longan polysaccharide improves the application in humoral immune function medicine in preparation, and carboxymethylation Arillus longan polysaccharide effective concentration is 40 ~ 160mg/kg.
(7) carboxymethylation Arillus longan polysaccharide strengthens the application in non-specific immune function medicine in preparation, and carboxymethylation Arillus longan polysaccharide effective concentration is 20 ~ 160mg/kg.
Beneficial effect of the present invention is:
The carboxymethylation Arillus longan polysaccharide of synthetic is used for immunocompetence experiment in antioxidation in vitro and body, obtain the experimental result of carboxymethylation Arillus longan polysaccharide immunoregulation effect and antioxygenation, for carboxymethylation Arillus longan polysaccharide provides reference frame in the application of field of medicaments.
Accompanying drawing explanation
Fig. 1 is Infrared spectroscopy figure before and after the Arillus longan polysaccharide carboxymethylation modification described in embodiment, and wherein A line represents Arillus longan polysaccharide A, and B line represents carboxymethylation Arillus longan polysaccharide A.
Fig. 2 is the ultra-oxygen anion free radical removing experimental result schematic diagram described in embodiment.
Fig. 3 affects result schematic diagram to mouse spleen index for the Arillus longan polysaccharide A described in embodiment and carboxymethylation Arillus longan polysaccharide A.(wherein, n=10,
*p<0.05 or
*p<0.01vs endoxan group)
Fig. 4 affects result schematic diagram to mouse delayed allergy for the Arillus longan polysaccharide A described in embodiment and carboxymethylation Arillus longan polysaccharide A.(wherein, n=10,
*p<0.05 or
*p<0.01vs endoxan group)
Fig. 5 is for the Arillus longan polysaccharide A described in embodiment and carboxymethylation Arillus longan polysaccharide A is to hemolysin (HC in mice serum
50) affect result schematic diagram.(wherein, n=10,
*p<0.05 or
*p<0.01vs endoxan group)
Fig. 6 affects result schematic diagram to LZM in mouse spleen for the Arillus longan polysaccharide A described in embodiment and carboxymethylation Arillus longan polysaccharide A.(wherein, n=10,
*p<0.05 or
*p<0.01vs endoxan group)
Fig. 7 affects result schematic diagram to LZM in mice serum for the Arillus longan polysaccharide A described in embodiment and carboxymethylation Arillus longan polysaccharide A.(n=10,
*p<0.05 or
*p<0.01vs endoxan group)
Embodiment
By the following examples technical scheme of the present invention is described further.
1, the preparation of Arillus longan polysaccharide A
Longan dry fruit is peeled off after stoning, takes 1.5kg longan aril in 4 times of water gagings, and controlling bath temperature is 90 DEG C, extracts 4h, extraction time 3 times at every turn.Alcohol precipitation after filtrate is concentrated, dry longan aril Crude polysaccharides.Longan aril Crude polysaccharides obtains longan aril refined polysaccharide after deproteinated, desolventing technology.DEAE-cellulose column (2.5cm × 80cm) grading purification longan aril refined polysaccharide, use water successively, 0.125mol/LNaCl, 0.30mol/LNaOH solution carries out wash-out, choose NaCl flow point, be further purified with SephacrylS-300HR post (2.0cm × 40cm), wash with water (4s/ drips), collect maximum enrichment peak and concentrate postlyophilization, obtain Arillus longan polysaccharide A.
2, Arillus longan polysaccharide A carboxymethylation is modified
Accurately take 0.2000g Arillus longan polysaccharide A sample, be dissolved in 10ml3mol/L sodium hydroxide solution, under vigorous stirring, slowly add Monochloro Acetic Acid, make its final concentration be 1.2mol/L.After continuing stirring reaction 3.2h after mixing at 73 DEG C, cooling, is adjusted to neutrality with the HCl of 2.0mol/L.Reaction solution is placed in molecular weight cut-off be 3500 dialysis tubing to dialyse 48h, dialyzate, through concentrated, obtains carboxymethylation Arillus longan polysaccharide A after alcohol precipitation and lyophilize.
3, Infrared spectroscopy before and after Arillus longan polysaccharide carboxymethylation
Dry carboxymethylation Arillus longan polysaccharide A1mg and 100mgKBr powder grind evenly in agate mortar, are pressed into thin slice, put into sample chamber, with Arillus longan polysaccharide A for contrast, in the infrared transform spectrometer of Fourier at 4000 ~ 400cm
-1carry out scanning in scope and obtain infrared absorpting light spectra.
As shown in Figure 1,3600-3200cm
-1the absorption peak in region is the charateristic avsorption band of sugar, is caused by the stretching vibration of hydroxyl O-H, shows to there is intermolecular and intramolecular hydrogen bond.3000-2800cm
-1the absorption peak in region is also the charateristic avsorption band of sugar, is caused by the stretching vibration of methyne C-H, shows there is-CH
2base exists.Spectrogram B 1603,1420,1326cm
-1there is the charateristic avsorption band of COO-in place, wherein, and 1603cm
-1place's absorption peak is strong peak, is the asymmetric stretching vibration of C-O key; 1420,1326cm
-1locate two peaks be in strong peak, be respectively the angle vibration of-CH-and the symmetrical stretching vibration peak of-COO.Demonstrate the certain carboxymethylation of Arillus longan polysaccharide A.After carboxymethylation, other charateristic avsorption bands of Arillus longan polysaccharide A there is no large change, and this modifies to carboxymethylation the evidence not changing Arillus longan polysaccharide A structure.
4, the mensuration of carboxymethylation Arillus longan polysaccharide A relative molecular weight
The mensuration of relative molecular weight adopts dextran gel filtration method.After SephadexG-100 gel column is loaded, balance 24h, prepares the solution of 3mg/ml by blue dextran 2000, loading calculates elution volume V
0, dextran standard T-10, T-40, T-70, T-500 (molecular weight is respectively 10KDa, 40KDa, 70KDa, 500KDa) are mixed with the solution of concentration 3mg/ml, respectively loading, its elution volume of Accurate Determining.And with Ve/V
0map with the logarithm lgM of relative molecular weight, obtain a typical curve.By the polysaccharide sample chromatography under the same conditions of unknown molecular amount, on typical curve, try to achieve its relative molecular mass according to elution volume.
Experiment show that Relative molecular weight markers equation is: y=-1.7448x+19.195, R
2=0.9915.The relative molecular weight being calculated carboxymethylation Arillus longan polysaccharide A by standard equation is: 1.51 × 10
5da.
5, the mensuration of carboxymethylation Arillus longan polysaccharide A substitution value
Method
(1) preparation of typical curve
Precision takes 0.2500g oxyacetic acid standard substance, with constant volume after water dissolution in 100ml volumetric flask, makes the solution that concentration is 2.50mg/ml.Accurate absorption standard solution 0.4,0.5,0.6,0.7,0.8,0.9,1.0ml, be placed in 25mL color-comparison tube respectively.Distilled water is mended to 1ml.Respectively add chromotropic acid solution 5ml and dense H
2sO
41ml, heats 30min after shaking up on boiling water bath.Treat that colorimetric cylinder is chilled to room temperature, drip 30%NH
4ac solution is to 25ml groove, and jolting, solution transition is red-purple.On 721 type spectrophotometers, at wavelength 570nm place, be reference with reagent blank, measure the absorbancy of each solution.Take absorbance as X-coordinate, oxyacetic acid mass concentration is ordinate zou, draws oxyacetic acid mass concentration typical curve.Equation of linear regression is: c=128.923A-1.044R
2=0.9955, within the scope of 32.0 ~ 80.0 μ g/ml, oxyacetic acid mass concentration and absorbance are linearly good.
(2) mensuration of polysaccharide sample carboxymethylation substitution value
Precision takes 0.1000g carboxymethylation Arillus longan polysaccharide, and after dissolving with appropriate distilled water, constant volume is in 10ml volumetric flask.Be made into the solution that concentration is 10mg/ml.Accurate absorption 0.5ml polysaccharide soln, by the method in 5 (1) items, measures light absorption value at wavelength 570nm place.Obtain according to working curve the quality that carboxymethylation Arillus longan polysaccharide A is equivalent to oxyacetic acid, the substitution value method of calculation of carboxymethylation Arillus longan polysaccharide A are:
DS＝l62B/[76-(59-1)B]
In above formula, B is the quality that every gram of carboxymethylation Arillus longan polysaccharide A sample is equivalent to oxyacetic acid.
76-oxyacetic acid molar mass (g/mol)
59-CH
2cOOH molar mass (g/mol)
1-hydrogen atom molar mass (g/mol)
Result
After measured, the degree of substitution by carboxymethyl of carboxymethylation Arillus longan polysaccharide A is 1.053.
6, Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A is to the mensuration of ultra-oxygen anion free radical Scavenging activity
Method
Getting 50mmol/LpH is that the Tris-HCl damping fluid 4.6ml of 8.2 is in test tube, after putting 25 DEG C of water-bath preheating 20min, add the test liquid 1ml of different concns, 10mmol/L pyrogallol 0.4ml, shake up rear 25 DEG C of water-bath 4min, use 8mol/LHCl0.1ml termination reaction immediately, with the Tris-HCl damping fluid zeroising of pH=8.2, and measure absorbancy (A at 325nm
x), model control group absorbancy (A
0) replace test liquid to measure with redistilled water 1ml, sample control group absorbancy (A
c) replace 10mmol/L Pyrogallol determination with redistilled water 0.4ml.The method of calculation of clearance rate E are as follows:
E＝[1-(A
x-A
c)/A
0]×100％
Result
As shown in Figure 2, the Arillus longan polysaccharide A before and after carboxymethylation all has the ability removing ultra-oxygen anion free radical, and presents dose-dependent effect.When mass concentration is identical, the Scavenging activity of the Arillus longan polysaccharide A after carboxymethylation is higher than the Arillus longan polysaccharide A of unmodified.When mass concentration is 12g/L, carboxymethylation Arillus longan polysaccharide A clearance rate is 62.9%, improves 29.4% than Arillus longan polysaccharide A.This is because the mechanism removing superoxide anion is relevant to the bond energy of O-H key, more electron-withdrawing group (as hydroxyl, carboxyl) is there is in carboxymethylation Arillus longan polysaccharide A molecular structure, O-H key more easily ruptures, can provide more proton in and superoxide anion.
7, Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A is on the impact of the spontaneous lipid peroxidation of mouse liver even slurry.
Method
Kunming mouse (25 ~ 30g, male and female half and half) de-cervical vertebra is put to death after overnight fasting, rapid taking-up liver organization, put in ice-cold normal saline and clean surperficial residual blood, on borneol, 10% murine liver tissue homogenate is made with 4 DEG C of physiological saline, 4 DEG C of centrifugal 15min of 4000r/min, getting supernatant liquor, to put layer refrigerator for subsequent use.Get 10% liver homogenate 1.0ml in test tube, add different concns test liquid 1.0ml to mix, after 37 DEG C of water-bath temperature incubate 1.5h, add 10% trichoroacetic acid(TCA) 2ml, 0.67% thiobarbituricacidα-2ml mixes, take out after reacting 15min in boiling water bath, be cooled to room temperature, the centrifugal 15min of 4000r/min, gets supernatant liquor, with physiological saline zeroising, and measure absorbancy (A at 532nm place
x).Model control group (A
0) replace test liquid, sample control group (A with physiological saline 1ml
c) replace 10% liver homogenate with physiological saline 1ml.Each concentration group replicate(determination) 5 parts.Measurement result represents with lipid peroxidation inhibiting rate (I), inhibiting rate I and IC
50the method of calculation of (reaching drug level needed for 50% inhibiting rate) are as follows:
I＝[A
0-(A
x-A
c)]/A
0×100％
Take I as ordinate zou, polysaccharide or vitamins C log concentration are X-coordinate mapping, set up regression equation, calculate IC
50.
Result
On the impact of the spontaneous lipid peroxidation of mouse liver even slurry before and after table 1 Arillus longan polysaccharide A carboxymethylation
This experimental result shows, and Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A all effectively can suppress the generation of murine liver tissue homogenate mda (MDA), and in certain dose-effect relationship, demonstrates good anti-lipid peroxidation active.Compared with Arillus longan polysaccharide A, carboxymethylation Arillus longan polysaccharide A anti-lipid peroxidation effect is stronger, but both inhibition of peroxidations are all not as good as vitamins C.
8, Arillus longan polysaccharide A and carboxymethyl Arillus longan polysaccharide A is to H
2o
2the impact of the erythrocyte hemolysis of induction
Method
Kunming mice (20 ± 2g) eyeball blood sampling 1 ~ 2ml, join in the centrifuge tube containing heparin sodium, the centrifugal 10min of 1500r/min, discard supernatant liquor and obtain erythroprecipitin, 3 times are cleaned with ice-cold normal saline, the centrifugal 5min of 1500r/min, makes the red blood cell suspension of 0.5% (V/V).Get 1ml red cell suspension, add the test liquid 1ml of different concns, then add 0.2ml50mmol/LH
2o
2, after water-bath temperature incubates 1h at 37 DEG C, add 5ml normal saline dilution, the centrifugal 10min of 3000r/min, gets supernatant liquor, take physiological saline as blank, measures absorbancy (A at 415nm wavelength place
x).Model control group (A
0) replace test liquid, sample control group (A with physiological saline 1ml
c) replace 0.5% red blood cell suspension with physiological saline 1ml.Each concentration group replicate(determination) 5 parts.Measurement result represents with haemolysis inhibiting rate (I), I and IC
50method of calculation as follows:
I＝[A
0-(A
x-A
c)]/A
0×100％
Take I as ordinate zou, polysaccharide or vitamins C log concentration are X-coordinate mapping, set up regression equation, calculate IC
50.
Result
To H before and after table 2 Arillus longan polysaccharide A carboxymethylation
2o
2the impact of the erythrocyte hemolysis of induction
This experimental result shows, and Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A is to H
2o
2induction erythrocyte hemolysis has certain restraining effect, and strengthens along with the increase restraining effect of polysaccharide concentration.The IC of Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A
50be respectively 620.82 and 262.76 μ g/ml, the latter suppresses H
2o
2the ability of induction erythrocyte hemolysis is outstanding.
9, endoxan causes the foundation of immunosuppression mouse model
After mouse adaptability is raised 24h, by body weight random packet, often organize 10, male and female half and half, except blank group intraperitoneal injection of saline, endoxan suppression group and all the other each experimental mice are every three days intraperitoneal injection of cyclophosphamide once, 20mg/kg, capacity 10ml/kg.Experiment component is Arillus longan polysaccharide A group (being divided into 160mg/ml dosage group, 80mg/kg dosage group, 40mg/kg dosage group and 20mg/kg dosage group), carboxymethylation Arillus longan polysaccharide A group (being divided into 160mg/ml dosage group, 80mg/kg dosage group, 40mg/kg dosage group and 20mg/kg dosage group) and lentinan group (40mg/kg).Each experimental group all takes gastric infusion, the blank group of physiological saline giving equivalent, and administration capacity is 10ml/kg, and every day is administered once, and experiment time-histories is 21 days.
10, index and spleen index measures:
Method
24h after last administration, takes off cervical vertebra place post mortem mouse and wins spleen, washing away bloodstain with physiological saline by mouse, weigh after sucking surface-moisture with filter paper, spleen is placed on ice after weighing immediately, treats post-processed.Calculate index and spleen index as follows:
Index and spleen index (mg/g)=W
1/ W
0
Wherein: W
1for Mouse Weight (g), W
0for spleen weight (mg).
Result
As can be seen from Figure 3, compared with Normal group, endoxan model group spleen index significantly reduces (p<0.05), and endoxan immunosuppression model modeling success is described.Compared with endoxan model group, the Arillus longan polysaccharide A of positive drug lentinan and various dose and carboxymethylation Arillus longan polysaccharide A all can improve the index and spleen index of immunosuppressed mice significantly and present dose-dependent effect (p<0.05 or p<0.01).Illustrate that Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A can promote that the immune organ of atrophing state recovers, improve immunity of organisms to a certain extent.
11, Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A is on the impact of mouse delayed type hypersensitivity (DTH)
Method
At the 16th day of administration, mouse web portion cuts off fritter hair, and area is about 3cm × 3cm, is kept flat upward by mouse web portion, draw 30 μ l1% dinitrofluorobenzene (DNFB) with liquid-transfering gun and slowly drop to unhairing position at twice, carry out initial immunity (sensitization).In 20th day evening upon administration, evenly embrocate mouse right ear both sides with 1% dinitrofluorobenzene 20 μ l, left ear is not wiped in contrast.24h after last administration, puts to death mouse cervical dislocation, cuts left and right auricle, take off the auricle that diameter is 6mm, weigh with punch tool, and mice ear degree is used for representing the degree of DTH.
Ear swelling degree (%)=(W
1-W
0)/W
0× 100%
Wherein: W
0for left ear weight, W
1for auris dextra weight.
Result
As can be seen from Figure 4, endoxan model group ear swelling degree is significantly compared (p<0.05) lower than Normal group, shows that mouse immune inhibition is successfully established.Compared with model group, positive lentinan group (40mg/kg), Arillus longan polysaccharide A group (40 ~ 160mg/kg) and carboxymethylation Arillus longan polysaccharide A group (20 ~ 160mg/kg) all can improve because endoxan suppresses the ear swelling degree (p<0.05 or p<0.01) that causes significantly, along with the increase of dosage, ear swelling degree increases thereupon, presents dose-dependence.Thus show that Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A can strengthen the T lymphocyte immunologic function of immunosuppressed mice.
12, Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A is to mice serum hemolysin (HC
50) impact
Method
Administration the 15th day, every mouse peritoneal is injected 2% sheep red blood cell (SRBC) (SRBC, SheepRedBloodCell) 0.2ml and is carried out immunity.Stop eating after last abdominal injection and do not cut off the water.Win eyeball after last administration 24h and get blood.Blood sample room temperature leaves standstill 2h, and 3000r/min is centrifugal, and 10min prepares serum.Get supernatant liquor 50 μ l, add in the test tube containing 2%SRBC and each 250 μ l of 10% guinea pig serum subsequently, mixing is testing sample; Take physiological saline as blank.Be incubated 30min in 37 DEG C of water-baths after, ice-water bath termination reaction, the centrifugal 5min of 1500r/min, gets supernatant liquor 250 μ l, measures testing sample optical density(OD) (OD) value by microplate reader in 540nm wavelength place.
Sample HC
50=(the OD of sample
540the OD of value-blank sample
540oD during)/SRBC HD50
540value
Result
As seen from Figure 5, hemolysin level (HC in endoxan group mice serum
50) significantly lower than Normal group (p<0.05), illustrate that mouse immune inhibition is successfully established.Compared with model group, positive lentinan group (40mg/kg), Arillus longan polysaccharide A group (80 ~ 160mg/kg) and carboxymethylation Arillus longan polysaccharide A group (40 ~ 160mg/kg) improve hemolysin level (p<0.05 or p<0.01) in immunosuppressed mice body all significantly, and this change presents dose-dependence.These results suggest that Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A recovers there is promoter action to the humoral immune function of immunosuppressed mice within the scope of finite concentration.
13, Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A is on the impact of mouse N,O-Diacetylmuramidase (LZM)
Method
Mouse spleen takes off rear physiological saline and prepares tissue homogenate by 1:10 (g/v), 5000r/min centrifuging and taking supernatant, and measure LZM content in spleen tissue and serum with commercial kit, concrete operations by specification carries out.
Result
As can be seen from Fig. 6 and Fig. 7, in endoxan group mouse spleen and serum, LZM content is significantly lower than Normal group (p<0.01), illustrates that mouse immune inhibition is successfully established.Compared with model group, positive lentinan group (40mg/kg), Arillus longan polysaccharide A group (40 ~ 160mg/kg) and carboxymethylation Arillus longan polysaccharide A group (20 ~ 160mg/kg) improve LZM level (p<0.05 or p<0.01) in immunosuppressed mice spleen and serum all significantly, and this change presents dose-dependence.These results suggest that Arillus longan polysaccharide A and carboxymethylation Arillus longan polysaccharide A recovers there is promoter action to the non-specific immune function of immunosuppressed mice within the scope of finite concentration.
Claims (10)
1. a carboxymethylation Arillus longan polysaccharide, it is characterized in that, described carboxymethylation Arillus longan polysaccharide is synthesized by following methods: be dissolved in by Arillus longan polysaccharide in sodium hydroxide solution, stirring reaction while adding Monochloro Acetic Acid, then reaction solution carried out dialysis is concentrated, obtain carboxymethylation Arillus longan polysaccharide after alcohol precipitation and lyophilize.
2. carboxymethylation Arillus longan polysaccharide according to claim 1, it is characterized in that, the synthetic method of described carboxymethylation Arillus longan polysaccharide is specially: be dissolved in by Arillus longan polysaccharide in 3mol/L sodium hydroxide solution, slowly add Monochloro Acetic Acid under vigorous stirring, makes its final concentration be 1.2mol/L; After continuing stirring reaction 3.2h after mixing at 73 DEG C, cooling, by HCl solution adjust ph to neutral; Reaction solution is placed in molecular weight cut-off be 3500 dialysis tubing to dialyse 48h, dialyzate, through concentrated, obtains carboxymethylation Arillus longan polysaccharide after alcohol precipitation and lyophilize.
3. the application of carboxymethylation Arillus longan polysaccharide in pharmacy.
4. carboxymethylation Arillus longan polysaccharide is preparing the application in anti-oxidation medicine or immunoregulation druge.
5. carboxymethylation Arillus longan polysaccharide is preparing the application in scavenging free radicals medicine, and the application effective concentration of carboxymethylation Arillus longan polysaccharide is 2 ~ 12mg/ml.
6. carboxymethylation Arillus longan polysaccharide is preparing the application of anti peroxidation of lipid medicine, and the application effective concentration of carboxymethylation Arillus longan polysaccharide is 0.25 ~ 4.00g/L.
7. carboxymethylation Arillus longan polysaccharide suppresses the application of erythrocyte hemolysis medicine in preparation, and the application effective concentration of carboxymethylation Arillus longan polysaccharide is 28.60 ~ 457.1 μ g/ml.
8. carboxymethylation Arillus longan polysaccharide improves the application in the horizontal medicine of T lymphocyte immunity in preparation, and the application effective concentration of carboxymethylation Arillus longan polysaccharide is 20 ~ 160mg/kg.
9. carboxymethylation Arillus longan polysaccharide improves the application in humoral immune function medicine in preparation, and the application effective concentration of carboxymethylation Arillus longan polysaccharide is 40 ~ 160mg/kg.
10. carboxymethylation Arillus longan polysaccharide strengthens the application in non-specific immune function medicine in preparation, and the application effective concentration of carboxymethylation Arillus longan polysaccharide is 20 ~ 160mg/kg.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| CN106916497A (en) * | 2017-03-28 | 2017-07-04 | 常州大学 | A kind of preparation method of the automobile water-based property metal flash paint of stably dispersing type |
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| CN109232754B (en) * | 2018-06-29 | 2021-07-13 | 桂林理工大学 | Passiflora edulis pericarp polysaccharide carboxymethylation modified product with improved antioxidant activity and preparation method thereof |
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| CN101579352A (en) * | 2009-05-27 | 2009-11-18 | 广西医科大学 | Application of polysaccharide component of longan pulp in pharmacy |
| CN102304190A (en) * | 2011-07-22 | 2012-01-04 | 天津实发中科百奥工业生物技术有限公司 | Preparation method of carboxymethyl levan and applications thereof |
| CN103030704A (en) * | 2012-12-05 | 2013-04-10 | 西北农林科技大学 | Method for preparing efficient oligomerized astragalus polysaccharide by carboxymethylated molecular modification |
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| CN101579352A (en) * | 2009-05-27 | 2009-11-18 | 广西医科大学 | Application of polysaccharide component of longan pulp in pharmacy |
| CN102304190A (en) * | 2011-07-22 | 2012-01-04 | 天津实发中科百奥工业生物技术有限公司 | Preparation method of carboxymethyl levan and applications thereof |
| CN103030704A (en) * | 2012-12-05 | 2013-04-10 | 西北农林科技大学 | Method for preparing efficient oligomerized astragalus polysaccharide by carboxymethylated molecular modification |
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| CN106916497A (en) * | 2017-03-28 | 2017-07-04 | 常州大学 | A kind of preparation method of the automobile water-based property metal flash paint of stably dispersing type |
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