CN103393711A - Applications of phosphate esterified longan pulp polysaccharide in pharmacy - Google Patents
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Abstract
The invention relates to applications of phosphate esterified longan pulp polysaccharide in pharmacy, and particularly relates to application in preparation of an immunomodulatory drug or an anti-oxidative drug, wherein the applications of the phosphate esterified longan pulp polysaccharide in preparation of the immunomodulatory drug relate to the following four aspects: application in preparation of a drug for enhancing the phagocytic ability of macrophage, application in preparation of a drug for enhancing the multiplication capacity of B leukomonocyte, application in preparation of a drug for enhancing the multiplication capacity of T leukomonocyte, and application in preparation of a drug for improving the IFN-gamma level in blood serum. According to the invention, the synthetic phosphate esterified longan pulp polysaccharide is applied to an active experiment on mice; an experiment result about the immune regulation effect and the antioxidant effect of the phosphate esterified longan pulp polysaccharide is obtained, thereby providing scientific evidences for application of the phosphate esterified longan pulp polysaccharide in the field of medicine.
Description
Technical field
The present invention relates to the application of Phosphation Arillus longan polysaccharide in pharmacy.
Background technology
Polysaccharide is little as a kind of toxic and side effects, the obvious natural immunity regulating drug of effect, scientists finds that bioactive polysaccharide has significant curative effect to cancer, immunologic derangement, diabetes, hepatic injury etc., utilization such as lentinus edodes polysaccharide injecta on antineoplaston, the utilization of Arillus longan polysaccharide oral liquid in immunomodulating, Angelica Polysaccharide capsule have become to treat respiratory repeated infection new drug and astragalin injection utilization in antiviral etc.It is few at occurring in nature, separating the Phosphation polysaccharide kind obtain, and most of Phosphation polysaccharide is to be isolated from microorganisms such as coccus, bacillus and yeast, and limited Phosphation polysaccharide, affected the research of people to its activity.Polysaccharide is carried out to the Phosphation structural modification, can change or give some polysaccharide compound special biological activity, can be more effectively by inducing the A549 apoptosis to suppress the propagation of people's lung cancer A549 cell than Inokopolyose as the Phosphation Inokopolyose; β-(1-3)-D-glucosan has been carried out after the phosphorylated, and its immunocompetence improves greatly.Fructose is carried out can be used for treating heavy cerebral infarction, adjuvant treatment of arrhythmias and antagonism gentamicin nephrotoxicity etc. after Phosphation.Therefore the physiological function of studying various polysaccharide and Phosphation derivant thereof just seems particularly necessary, therefrom can continue to optimize the method for Phosphation, the biological activity of research Phosphation polysaccharide, such as the phosphate radical substitution value on the binding site of the impact of pharmacologically active, activity change that spatial variations causes, the phosphate radical interactively to activity, can also be from the receptor of Molecular level study Phosphation polysaccharide effect, by round pcr, study acceptor gene, analyze activated protein and cytokine etc.From the physiologically active aspect, design better Studies on Phosphorylation of Polysaccharides derivant, the medicine resource of how developable antitumor and raising immunity of organism also is provided for clinical practice simultaneously.
Summary of the invention
The purpose of this invention is to provide the application of Phosphation Arillus longan polysaccharide in pharmacy, obtain to regulate in Phosphation Arillus longan polysaccharide body the result of immunization and antioxidation.
The present invention achieves the above object by the following technical programs:
The application of Phosphation Arillus longan polysaccharide in pharmacy, the especially application in preparation immunomodulating or anti peroxidation of lipid medicine.
Further, the application of Phosphation Arillus longan polysaccharide in preparing immunoregulation medicament comprises aspect following 4:
(1) application of Phosphation Arillus longan polysaccharide in the phagocytic activity medicine of preparation enhancing macrophage, Phosphation Arillus longan polysaccharide valid density is 40～160mg/kg;
(2) the Phosphation Arillus longan polysaccharide strengthens the application in bone-marrow-derived lymphocyte multiplication capacity medicine in preparation, and Phosphation Arillus longan polysaccharide valid density is 160mg/kg;
(3) the Phosphation Arillus longan polysaccharide strengthens the application in the lymphocytic multiplication capacity medicine of T in preparation, and Phosphation Arillus longan polysaccharide valid density is 160mg/kg;
(4) application in the horizontal medicine of Phosphation Arillus longan polysaccharide IFN-γ in preparation raising serum, Phosphation Arillus longan polysaccharide valid density is 160mg/kg.
The Phosphation Arillus longan polysaccharide improves the application in anti peroxidation of lipid ability medicine in preparation, and Phosphation Arillus longan polysaccharide valid density is 40～160mg/kg.
Beneficial effect of the present invention is:
The Phosphation Arillus longan polysaccharide of synthetic, for the mice intracorporeal active experiment, is obtained to the experimental result of Phosphation Arillus longan polysaccharide immunoregulation effect and antioxidation, for the Phosphation Arillus longan polysaccharide, in the application of field of medicaments, provide scientific basis.
The specific embodiment
By the following examples technical scheme of the present invention is described further.
1, acute toxicity testing
Laboratory animal: 20 kunming mices, SPF level, male and female half and half, body weight 18～22g.Fasting be can't help by body weight, being divided at random 5 groups (blank group, 5mg/kg dosage group, 50mg/kg dosage group, 500mg/kg dosage group, 5000mg/kg dosage groups), 4 every group (male and female half and half) after water 12h.Gavage gives each tested medicine group mice, and the administration capacity is 10ml/kg, and blank group gavage gives distilled water, and in one day, gavage once.Primary part observation in administration 4h, repeatedly observe in 24h, and regularly observe twice later every day, continuous 7 days.Relatively, feed drinking-water, stress, the mental status and Excreta etc. are showed no obvious abnormalities, and dead mouse also do not occur for result demonstration, the mice of each dosage group and blank group.According to the content of " chemicals toxicity authenticate technology standard " acute oral toxicity test, adopt 5000mg/kg dosage, as do not cause animal dead, can no longer carry out the acute oral toxicity test of a plurality of dosage.Illustrate that the Phosphation Arillus longan polysaccharide is low toxicity compounds.
2, cyclophosphamide causes the foundation of immunosuppression mouse model
After the mice adaptability is fed to 24h, by body weight, be divided at random 8 groups, 10 every group, male and female half and half, except blank group intraperitoneal injection of saline, inhibition group and all the other each experimental mice every three days intraperitoneal injection of cyclophosphamide once, 20mg/kg, capacity 10ml/kg.Experiment component is Phosphation Arillus longan polysaccharide high dose group (160mg/kg), dosage group (80mg/kg) in the Phosphation Arillus longan polysaccharide, Phosphation Arillus longan polysaccharide low dose group (40mg/kg), positive controls is Arillus longan polysaccharide group (80mg/kg), pidotimod group (200mg/kg), lentinan group (80mg/kg).Each experimental group is all taked gastric infusion, and blank group gives the normal saline of equivalent, and the administration capacity is 10ml/kg, and be administered once every day, and the experiment time-histories is 21 days.
3, peritoneal macrophage is engulfed the dimethyl diaminophenazine chloride experiment
Method
After eyeball of mouse is got blood, with after 75% soak with ethanol 2min, to the aseptic PBS liquid of mouse peritoneal injection 5ml, rub abdominal part 2min, with syringe pumpback peritoneal fluid, avoid abdominal viscera as far as possible, avoid injured blood vessel to cause bleeding, the centrifugal 15min of peritoneal fluid 2000rpm/min, abandon supernatant, precipitation adds the PRMI-1640 culture fluid that contains 10% calf serum, gently piping and druming, cell is mixed, cell number is transferred to 1 * 10
6/ ml, be inoculated in 96 orifice plates, every hole 100 μ l, and at 37 ℃, 5%CO
2Under condition, cultivate 4h, inhale and to abandon whole culture fluid, remove not adherent macrophage, add 0.1% dimethyl diaminophenazine chloride dyestuff (it is formulated that the 0.2500g dimethyl diaminophenazine chloride joins the sodium chloride injection of 250ml0.9%), every hole 100 μ l, at 37 ℃, 5%CO
2Under condition, cultivate 30min again, carefully draw unnecessary neutral red solution, every hole is washed 3 times with PBS liquid, add cell pyrolysis liquid (acetic acid: the 200 μ l of dehydrated alcohol=1:1), vibration, standing gently, 4 ℃ are spent the night, and at λ=570nm place, survey the OD value, with the power of OD value representation macrophage ability.
Experimental result
Table 1 Phosphation Arillus longan polysaccharide is engulfed the impact of dimethyl diaminophenazine chloride ability on the immunocompromised Turnover of Mouse Peritoneal Macrophages
| Group | The OD value |
| The blank group | 0.168±0.10 |
| Cyclophosphamide inhibition group (CY) | 0.096±0.06 |
| CY+ Arillus longan polysaccharide group | 0.216±0.22* |
| CY+ lentinan group | 0.273±0.27* |
| CY+ pidotimod group | 0.384±0.38* |
| CY+ Phosphation Arillus longan polysaccharide high dose group | 0.352±0.35* |
| Dosage group in CY+ Phosphation Arillus longan polysaccharide | 0.251±0.25* |
| CY+ Phosphation Arillus longan polysaccharide low dose group | 0.190±0.19* |
(note: * is less than 0.05, to be significantly with inhibition group comparison P value)
The experimental result of table 1 gained shows: as the macrophage of nonspecific immunity defence and immunne response, in the very first time, cyclophosphamide is caused to the immunoregulation effect that the immunocompromised mice risen apparent in view, wherein the ability of pidotimod group macrophage phagocytic dimethyl diaminophenazine chloride is the highest, the phagocytic activity of each experimental group macrophage all obviously improves, and with the increase of dosage, the phagocytic activity of macrophage is in rising trend.
4, the impact (mtt assay) of Phosphation Arillus longan polysaccharide on immunocompromised mice B and T lymphproliferation response
Method
(1) preparation of MTT solution: 0.5000g MTT is dissolved in the aseptic PBS liquid of 100ml, keeps in Dark Place.
(2) preparation of canavaline (ConA) solution: the ConA of 0.40mg is dissolved in the PRMI-1640 culture fluid that 20.00ml do not contain calf serum, and being made into concentration is the ConA solution of 20 μ g/ml.
(3) preparation of lipopolysaccharide (LPS) solution: the LPS of 0.60mg is dissolved in the PRMI-1640 culture fluid that 30.00ml do not contain calf serum and is made into the LPS solution that concentration is 30 μ g/ml.
(4) mtt assay detects the bone-marrow-derived lymphocyte multiplication capacity: spleen is shredded to about 1mm with eye scissors
3, under the condition of 37 ℃, the pancreatin with 0.25% digests the spleen tissue that shreds, the tissue that has digested is with the PRMI-1640 culture fluid that contains 10% calf serum of 2 times of volumes, piping and druming mixes gently, makes the pancreatin inactivation, reduces the impact of pancreatin on living cells, cross 200 order cell sieves, the centrifugal 8min of filtrate 1000rpm/min, abandon supernatant, and precipitation is spleen cell, add the PRMI-1640 culture fluid to be prepared into single splenocyte suspension, cell concentration is adjusted to 2 * 10
6/ ml, add 96 orifice plates, and then every hole 100 μ l add each 100 μ l of LPS of 30 μ g/ml.Each sample is established 3 repeating holes.Put 37 ℃, 5%CO
2In incubator, cultivate 66h.Every hole adds 20 μ l MTT, 37 ℃ hatch 4h after, the careful suction abandoned supernatant, then adds 150 μ l dimethyl sulfoxide (DMSO) vibrations.In microplate reader, survey OD value, λ=490nm.Take separately without the OD value of LPS stimulating group as 100%, all the other groups all are converted to percentage ratio and do data statistics processing.
(5) mtt assay detects T lymphopoiesis ability: use the ConA stimulus object instead, all the other operate with step 4.
Result
The impact of table 2 Phosphation Arillus longan polysaccharide on immunocompromised mouse spleen bone-marrow-derived lymphocyte multiplication capacity
(note: * is less than 0.05, to be significantly with inhibition group comparison P value,
sFor with the blank group, comparing p less than 0.05)
By experimental result measured in table 2, shown, Phosphation Arillus longan polysaccharide experimental group is along with the increase of dosage, and the stimulating ring phosphamide causes immunocompromised mouse spleen bone-marrow-derived lymphocyte multiplication capacity also to be strengthened.With the blank group, contrast, only have pidotimod group, Phosphation Arillus longan polysaccharide high dose group to have significant difference on to the bone-marrow-derived lymphocyte multiplication capacity, each experimental group all can recover to a certain extent or strengthen cyclophosphamide and cause immunocompromised mice bone-marrow-derived lymphocyte multiplication capacity.
The impact of table 3 Phosphation Arillus longan polysaccharide on immunocompromised mouse spleen T lymphopoiesis ability
(note: * is less than 0.05, to be significantly with inhibition group comparison P value,
sFor with the blank group, comparing p less than 0.05)
By experimental result measured in table 3, shown, in positive controls, the pidotimod group is the highest on the impact that cyclophosphamide causes immunocompromised mouse spleen T lymphopoiesis ability, stimulation index reaches 165%, the Arillus longan polysaccharide group is taken second place, stimulation index reaches 136%, and the lentinan group is minimum, and with respect to inhibition group there was no significant difference.Phosphation Arillus longan polysaccharide group is along with the increase of dosage, and the stimulation that cyclophosphamide is caused to immunocompromised mouse T lymphocyte multiplication capacity is also stronger, and irritation level and the normal group of low dose group are suitable, and the stimulation index of high dose group reaches 149%.
5, the impact of Phosphation Arillus longan polysaccharide on the Cytokine of Serum IFN γ content of immunocompromised mice
Method
Win eyeball of mouse and get blood, be placed in the EP pipe of 2ml, 4 ℃ of centrifugal 10min of lower 3500rpm/min, draw the serum of upper strata clarification, adopts the ELISA test kit, for measuring the content of serum IFN γ.
Result
The impact of table 4 Phosphation Arillus longan polysaccharide on IFN γ content in the immunocompromised mice serum
| Group | A 450nm | IFN γ concentration (pg/ml) |
| Cyclophosphamide inhibition group | 0.322±0.026 | 66.270 |
| The blank group | 0.365±0.044 | 105.009* |
| CY+ Arillus longan polysaccharide group | 0.359±0.041 | 100.003* |
| CY+ lentinan group | 0.360±0.037 | 100.630* |
| CY+ pidotimod group | 0.347±0.027 | 88.369 |
| CY+ Phosphation Arillus longan polysaccharide high dose group | 0.361±0.040 | 101.191* |
| Dosage group in CY+ Phosphation Arillus longan polysaccharide | 0.343±0.043 | 85.166 |
| CY+ Phosphation Arillus longan polysaccharide low dose group | 0.329±0.041 | 72.727 |
(note: * is less than 0.05, to be significantly with inhibition group comparison P value)
As shown in table 4, experimental result shows: the Phosphation Arillus longan polysaccharide is with the increase of dosage, and IFN γ content is in rising trend, and the IFN γ content of high dose group reaches 101.191pg/ml, far above the inhibition group, significant difference is arranged, but a little less than the normal group level.
6, the impact of Phosphation Arillus longan polysaccharide on immunocompromised Mouse Liver lipid peroxidation suppression ratio
Method
(1) take the weight of every mouse liver, in the ratio of W:V=1:10, add the normal saline of pH=7.4, fully grind and be made into 10% liver homogenate, the centrifugal 10min of 4000rpm/min, get supernatant and be put in-20 ℃ of Refrigerator stores standby.
(2) the liver homogenate 2.00ml that draws each experimental group 10% is in tool plug test tube, and normal saline adds respectively 10% trichloroacetic acid solution 4.00ml as a control group, and 0.67% 2-thiobarbituricacidα-2.0ml, mix boiling water bath 15min on eddy blending machine.
(3) after natural cooling, the centrifugal 5min of 3500rpm, get supernatant.
(4) in the 400-720nm wave-length coverage, measure maximum absorption wavelength.
(5) take maximum absorption wavelength as incident illumination, measure the light absorption value of each experimental group reactant liquor, every group of horizontal survey 3 times, average.
(6) the Phosphation Arillus longan polysaccharide is calculated as follows immunocompromised Mouse Liver lipid peroxidation suppression ratio:
Result
The impact of table 5 Phosphation Arillus longan polysaccharide on immunocompromised lipid peroxidation of mice suppression ratio
| Group | The OD value | Suppression ratio % |
| The blank group | 0.546±0.08* | - |
| Cyclophosphamide inhibition group | 0.789±0.15 | - |
| CY+ Arillus longan polysaccharide group | 0.628±0.04* | 20.4 |
| CY+ lentinan group | 0.587±0.07* | 25.6 |
| CY+ pidotimod group | 0.721±0.13 | 8.6 |
| The CY+LYP2-P high dose group | 0.421±0.04* | 46.6 |
| Dosage group in CY+LYP2-P | 0.456±0.07* | 42.2 |
| The CY+LYP2-P low dose group | 0.517±0.06* | 34.5 |
(note: * is less than 0.05, to be significantly with inhibition group comparison P value)
As shown in table 5, the OD value of cyclophosphamide inhibition group is the highest, illustrates that in the liver of inhibition group mice, MDA content is the highest.Positive control pidotimod and lentinan play the antioxidation effect to the immunocompromised mice and are weaker than the Phosphation Arillus longan polysaccharide, the lipid peroxidation suppression ratio of Phosphation Arillus longan polysaccharide all strengthens gradually along with the increase of dosage, and the suppression ratio of high dose group can reach 46.6%.
Claims (7)
1. the application of Phosphation Arillus longan polysaccharide in pharmacy.
2. the application of Phosphation Arillus longan polysaccharide according to claim 1 in pharmacy, is characterized in that, described pharmacy refers to and prepares immunoregulation medicament or anti-oxidation medicine.
3. the application of Phosphation Arillus longan polysaccharide according to claim 2 in pharmacy, it is characterized in that, described immunoregulation medicament is for strengthening the medicine of macrophage phagocytic ability, and during application, Phosphation Arillus longan polysaccharide valid density is 40～160mg/kg.
4. the application of Phosphation Arillus longan polysaccharide according to claim 2 in pharmacy, is characterized in that, described immunoregulation medicament is for strengthening the medicine of bone-marrow-derived lymphocyte multiplication capacity, and during application, Phosphation Arillus longan polysaccharide valid density is 160mg/kg.
5. the application of Phosphation Arillus longan polysaccharide according to claim 2 in pharmacy, is characterized in that, described immunoregulation medicament is for strengthening the medicine of T lymphopoiesis ability, and during application, Phosphation Arillus longan polysaccharide valid density is 160mg/kg.
6. the application of Phosphation Arillus longan polysaccharide according to claim 2 in pharmacy, is characterized in that, described immunoregulation medicament is for improving the medicine of IFN-γ level in serum, and during application, Phosphation Arillus longan polysaccharide valid density is 160mg/kg.
7. the application of Phosphation Arillus longan polysaccharide according to claim 2 in pharmacy, is characterized in that, during described anti-oxidation medicine application, Phosphation Arillus longan polysaccharide valid density is 40～160mg/kg.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105030625A (en) * | 2015-09-17 | 2015-11-11 | 天峨县平昌生态农业有限公司 | Chloasma-removing beautifying mask |
| CN106265176A (en) * | 2015-05-19 | 2017-01-04 | 成都宝科生物科技有限公司 | A kind of face cosmetics |
| CN116370605A (en) * | 2023-05-15 | 2023-07-04 | 山东博森医学工程技术有限公司 | Medicine for improving proliferation and phagocytic capacity of immune cells |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102775513A (en) * | 2012-08-24 | 2012-11-14 | 广西医科大学 | Phosphated Arillus Longan polysaccharide and synthesis method thereof |
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| CN102775513A (en) * | 2012-08-24 | 2012-11-14 | 广西医科大学 | Phosphated Arillus Longan polysaccharide and synthesis method thereof |
Non-Patent Citations (1)
| Title |
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| 易阳等: "龙眼多糖的提取分离及其生物活性研究进展", 《广东农业科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106265176A (en) * | 2015-05-19 | 2017-01-04 | 成都宝科生物科技有限公司 | A kind of face cosmetics |
| CN105030625A (en) * | 2015-09-17 | 2015-11-11 | 天峨县平昌生态农业有限公司 | Chloasma-removing beautifying mask |
| CN116370605A (en) * | 2023-05-15 | 2023-07-04 | 山东博森医学工程技术有限公司 | Medicine for improving proliferation and phagocytic capacity of immune cells |
| CN116370605B (en) * | 2023-05-15 | 2023-09-12 | 山东博森医学工程技术有限公司 | Medicine for improving proliferation and phagocytic capacity of immune cells |
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