Nothing Special   »   [go: up one dir, main page]

CN106109487B - Antioxidant with anti-tumor effect and preparation method thereof - Google Patents

Antioxidant with anti-tumor effect and preparation method thereof Download PDF

Info

Publication number
CN106109487B
CN106109487B CN201610573418.0A CN201610573418A CN106109487B CN 106109487 B CN106109487 B CN 106109487B CN 201610573418 A CN201610573418 A CN 201610573418A CN 106109487 B CN106109487 B CN 106109487B
Authority
CN
China
Prior art keywords
stl
polysaccharide
antioxidant
tumor effect
lobayense heim
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201610573418.0A
Other languages
Chinese (zh)
Other versions
CN106109487A (en
Inventor
陈彦
王亚
程乃嘉
李雪慧
周礼元
蔡正楠
郭文强
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Anhui University
Original Assignee
Anhui University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Anhui University filed Critical Anhui University
Priority to CN201610573418.0A priority Critical patent/CN106109487B/en
Publication of CN106109487A publication Critical patent/CN106109487A/en
Application granted granted Critical
Publication of CN106109487B publication Critical patent/CN106109487B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • CCHEMISTRY; METALLURGY
    • C08ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08BPOLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Materials Engineering (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Polymers & Plastics (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)

Abstract

本发明公开了一种兼具抗肿瘤作用的新型抗氧化剂及其制备方法,其特征在于:该抗氧化剂为硫酸化修饰的金福菇多糖STL‑3,其是以金福菇多糖TL‑3为原料,利用三氧化硫‑吡啶法进行硫酸化修饰获得;该化合物分子量为8.12×103Da、硫酸根含量为13.60%、取代度为1.22,由鼠李糖、葡萄糖、半乳糖、葡萄糖醛酸、阿拉伯糖、甘露糖和半乳糖醛酸7种单糖组成。体外抗氧化实验表明STL‑3的清除能力(DPPH,·O2 ,·OH和还原力)已经超过了Vc;体外抗肿瘤实验显示STL‑3对HeLa细胞和MCF‑7有很强的抑制能力,且对HELF细胞无毒副作用;结果表明STL‑3将会在食品和医药保健等领域具有巨大的应用前景。The invention discloses a novel antioxidant with anti-tumor effect and a preparation method thereof, which is characterized in that: the antioxidant is sulfated modified chrysanthemum polysaccharide STL-3, which is based on the phlox polysaccharide TL-3 As a raw material, it is obtained by sulfation modification by sulfur trioxide-pyridine method; the compound has a molecular weight of 8.12×10 3 Da, a sulfate content of 13.60%, and a substitution degree of 1.22. It is composed of 7 monosaccharides, arabinose, mannose and galacturonic acid. In vitro anti-oxidation experiments showed that the scavenging ability of STL‑3 (DPPH, ·O 2‑ , ·OH and reducing power) had exceeded Vc; in vitro anti-tumor experiments showed that STL‑3 had a strong inhibitory effect on HeLa cells and MCF‑7 ability, and has no toxic and side effects on HELF cells; the results show that STL‑3 will have great application prospects in the fields of food and medical care.

Description

A kind of antioxidant and preparation method thereof having both antitumor action
Technical field
The present invention relates to a kind of deep processed products of edible mushroom, in particular to the polysaccharide extracted using T.lobayense Heim as raw material, warp The antioxidant for having both antitumor action that oversulfated modification generates.
Background technique
T.lobayense Heim (Tricholoma lobayense Heim) is also known as Tricholoma Giganteum Massee, big spoken parts in traditional operas mushroom, Tricholoma lobayense, Japan claims white matsutake, is that a kind of natural health care of preciousness and high-grade edible mushroom, polysaccharide and proteoglycans have Immunological regulation and antitumor activity.
Chinese invention patent ZL201210014537.4 shows that T.lobayense Heim polysaccharide TL-3 is the one kind extracted from T.lobayense Heim The acidic polysaccharose of hyperbranched small molecule, molecular weight are 4.22 × 103Da, have with the comparable antioxidant activity of Vc, can As a kind of Nantural non-toxic antioxidant, it is applied to the fields such as food, medicine and cosmetics.But the compound still has stabilization The disadvantages of property is poor, oxidation resistance is to be improved.
Currently, polysaccharide due to can improve its intrinsic bioactivity after chemical modification or obtain new functional characteristic by Extensive concern.Common method of modifying has sulphation, acetylation, carboxy methylation etc., wherein sulphation is the biology for enhancing polysaccharide Active classical way.Research shows that sulfated polysaccharides is the potent inhibitor of human immunodeficiency virus (HIV).Ganoderma lucidum polysaccharide More effectively inhibit the growth of S-180 cell after sulphation than before.The polysaccharide sulfated anti-oxidant work that they not only can be improved Property, also them is made to have effects that antibacterial.The method of sulphation modification has very much, such as chlorosulfonic acid method, sulphate method, wherein chlorine sulphur Acid meets water capacity explosive, and concentrated sulfuric acid corrosivity is too strong.
In view of reagent cost and the yield of the convenient of method, safety and product, to realize industrialized production, the present invention Sulfur trioxide-pyridine method is applied in T.lobayense Heim polysaccharide for the first time in the sulphation modification of the highest component TL-3 of activity, is obtained A kind of sulphation modification method simple, mild, product yield is high and active and strong stability.
Summary of the invention
It is an object of that present invention to provide a kind of novel antioxidant for having both antitumor action and preparation method thereof namely sulphur It is acidified the T.lobayense Heim polysaccharide STL-3 of modification, has both antitumor and antioxidation, and STL-3 is to HELF cytotoxic pair Effect has huge application potential in fields such as health cares;The present invention is for the first time by sulfur trioxide-pyridine (SO3- Py) method fortune It is a kind of simple, mild, product yield height and active and stability in the sulphation modification for using T.lobayense Heim polysaccharide TL-3 Strong sulphation modification method.
The present invention is to realize goal of the invention, is adopted the following technical scheme that
A kind of novel antioxidant having both antitumor action, it is characterised in that: the antioxidant is sulphation modification T.lobayense Heim polysaccharide STL-3, be using T.lobayense Heim polysaccharide TL-3 as raw material, using sulfur trioxide-pyridine method carry out sulphation repair Decorations obtain.The T.lobayense Heim polysaccharide STL-3 molecular weight of the sulphation modification is 8.12 × 103Da, sulfate radical content are 13.60%, degree of substitution 1.22, by rhamnose, glucose, galactolipin, glucuronic acid, arabinose, mannose and gala 7 kinds of monosaccharide compositions of uronic acid.
The preparation method of the above-mentioned novel antioxidant for having both antitumor action is: being not less than 95% T.lobayense Heim with purity Polysaccharide TL-3 is raw material, 35~50 times of material quality of solvent formamide (PF), 25 DEG C of water-bath 30min is added, then press raw material matter 1~2 times of addition sulfur trioxide-pyridine (SO of amount3- Py), magnetic agitation 3h under 60~80 DEG C of oil baths, finally with 4mol/L's NaOH tune pH dialyses, concentration, alcohol precipitation is overnight, and precipitating is freeze-dried to obtain T.lobayense Heim sulfated polysaccharides STL-3, as to neutrality Have both the novel antioxidant of antitumor action.
The beneficial effects of the present invention are embodied in:
The present invention utilizes sulfur trioxide-pyridine (SO using T.lobayense Heim polysaccharide TL-3 as raw material3- Py) method carry out sulphation repair Decorations, have obtained the T.lobayense Heim polysaccharide STL-3 of sulphation modification.This method step is easy, easily operated, and the conversion ratio of TL-3 is 30%, in product sulfate radical content be 13.60%, degree of substitution 1.22, final products purity is up to 99% or more.
Gained STL-3 of the invention is removing DPPH and O2 -The IC of free radical50Value has been over positive control Vitamin C Acid;4 times of TL-3 high of inhibiting rate ratio to HeLa cell of STL-3;STL-3 is to the inhibiting rate of MCF-7 cell almost 5 more than TL-3 Times;In the concentration of 10 μ g/mL, (a famous anticancer drug can largely make cancer cell lethal to 5-Fu, but simultaneously Also have huge side effect to normal cell) it is only 50% or so to the survival rate of HELF cell, but in 400 μ g/mL, STL- 4 times of 5-Fu high of survival rate ratio or more of 3, and have certain promotion growth to cell after modifying;STL-3 has both antitumor function Energy and oxidation resistance, obvious effect, market application prospect are very wide.
Detailed description of the invention
Fig. 1 is the T.lobayense Heim of the raw materials used T.lobayense Heim polysaccharide TL-3 of the embodiment of the present invention 1 and products obtained therefrom sulphation modification High performance liquid chromatography (HPLC) test map of polysaccharide STL-3;
Fig. 2 is the T.lobayense Heim of the raw materials used T.lobayense Heim polysaccharide TL-3 of the embodiment of the present invention 1 and products obtained therefrom sulphation modification The infared spectrum of polysaccharide STL-3;
Fig. 3 is sample in the embodiment of the present invention 4 to the test result of the Scavenging activity of DPPH free radical;
Fig. 4 is sample in the embodiment of the present invention 4 to superoxide anion (O2 -) Scavenging activity test result;
Fig. 5 is sample in the embodiment of the present invention 4 to the test result of the Scavenging activity of hydroxyl radical free radical (OH);
Fig. 6 is the test result of sample reducing power in the embodiment of the present invention 4;
Fig. 7 is sample in the embodiment of the present invention 4 to the test result of the rejection ability of HeLa and MCF-7 cell;
Fig. 8 is sample in the embodiment of the present invention 4 to the test result of the toxic effect to HELF cell.
Specific embodiment
Embodiment 1
The novel antioxidant that the present embodiment has both antitumor action is the T.lobayense Heim polysaccharide STL-3 of sulphation modification, Molecular weight is 8.12 × 103Da, sulfate radical content 13.60%, degree of substitution 1.22, by rhamnose, glucose, galactolipin, 7 kinds of glucuronic acid, arabinose, mannose, galacturonic acid monosaccharide compositions;Its preparation step is as follows:
It takes purity to be not less than 95% T.lobayense Heim polysaccharide TL-3 100mg, the solvent formamide (PF) of 5g, 25 DEG C of water is added 30min is bathed, the SO of 200mg is added3- Py, magnetic agitation 3h, NaOH (4mol/L) adjusts pH to neutrality under 70 DEG C of oil baths, dialysis, Concentration, alcohol precipitation is overnight, precipitates the freeze-dried T.lobayense Heim polysaccharide STL-3 for obtaining sulphation modification.
Fig. 1 is the T.lobayense Heim polysaccharide of the raw materials used T.lobayense Heim polysaccharide TL-3 of the present embodiment and products obtained therefrom sulphation modification High performance liquid chromatography (HPLC) test map of STL-3;Fig. 2 is the raw materials used T.lobayense Heim polysaccharide TL-3 of the present embodiment and gained produces The infared spectrum of the T.lobayense Heim polysaccharide STL-3 of product sulphation modification.
Embodiment 2
The novel antioxidant that the present embodiment has both antitumor action is the T.lobayense Heim polysaccharide STL-3 of sulphation modification, Molecular weight is 8.12 × 103Da, sulfate radical content 13.60%, degree of substitution 1.22, by rhamnose, glucose, galactolipin, 7 kinds of glucuronic acid, arabinose, mannose, galacturonic acid monosaccharide compositions;Its preparation step is as follows:
It takes purity to be not less than 95% T.lobayense Heim polysaccharide TL-3 100mg, the solvent formamide (PF) of 4g, 25 DEG C of water is added 30min is bathed, the SO of 100mg is added3- Py, magnetic agitation 3h, NaOH (4mol/L) adjusts pH to neutrality under 80 DEG C of oil baths, dialysis, Concentration, alcohol precipitation is overnight, precipitates the freeze-dried T.lobayense Heim polysaccharide STL-3 for obtaining sulphation modification.
The HPLC map and infared spectrum of the present embodiment products obtained therefrom are similar to Example 1.
Embodiment 3
The novel antioxidant that the present embodiment has both antitumor action is the T.lobayense Heim polysaccharide STL-3 of sulphation modification, Molecular weight is 8.12 × 103Da, sulfate radical content 13.60%, degree of substitution 1.22, by rhamnose, glucose, galactolipin, 7 kinds of glucuronic acid, arabinose, mannose, galacturonic acid monosaccharide compositions;Its preparation step is as follows:
It takes purity to be not less than 95% T.lobayense Heim polysaccharide TL-3 100mg, is added the solvent formamide (PF) of 4.5g, 25 DEG C Water-bath 30min adds the SO of 200mg3- Py, magnetic agitation 3h, NaOH (4mol/L) adjusts pH to neutrality under 60 DEG C of oil baths, thoroughly Analysis, concentration, alcohol precipitation is overnight, precipitates the freeze-dried T.lobayense Heim polysaccharide STL-3 for obtaining sulphation modification.
The HPLC map and infared spectrum of the present embodiment products obtained therefrom are similar to Example 1.
Embodiment 4
The novel antioxidant that the present embodiment has both antitumor action is the T.lobayense Heim polysaccharide STL-3 of sulphation modification, Molecular weight is 8.12 × 103Da, sulfate radical content 13.60%, degree of substitution 1.22, by rhamnose, glucose, galactolipin, 7 kinds of glucuronic acid, arabinose, mannose, galacturonic acid monosaccharide compositions;Its preparation step is as follows:
It takes purity to be not less than 95% T.lobayense Heim polysaccharide TL-3 100mg, is added the solvent formamide (PF) of 3.5g, 25 DEG C Water-bath 30min adds the SO of 200mg3- Py, magnetic agitation 3h, NaOH (4mol/L) adjusts pH to neutrality under 72 DEG C of oil baths, thoroughly Analysis, concentration, alcohol precipitation is overnight, precipitates the freeze-dried T.lobayense Heim polysaccharide STL-3 for obtaining sulphation modification.
The HPLC map and infared spectrum of the present embodiment products obtained therefrom are similar to Example 1.
The antioxidant activity of 4 products obtained therefrom of embodiment is tested:
(1) to the scavenging effect of DPPH free radical
Prepare the gold of various concentration (25 μ g/mL, 50 μ g/mL, 100 μ g/mL, 200 μ g/mL, 300 μ g/mL, 400 μ g/mL) The aqueous solution of the T.lobayense Heim polysaccharide STL-3 of good fortune mushroom polysaccharide TL-3 and sulphation modification, takes the methanol of 2mL and 2mL DPPH molten respectively Liquid (0.13mmol/L) mixes.Reaction mixture is uniformly mixed with quick vortex mixer and is incubated for 30min at 25 DEG C of room temperature, it is quiet It sets, measures light absorption value with ultra-violet and visible spectrophotometer at 517nm;Every group experimental setup three parallel, takes light absorption value Average value.Make positive control with Vc to operate instead of T.lobayense Heim polysaccharide by above-mentioned steps;
The Scavenging activity of DPPH free radical is calculated according to following formula:
Wherein, A0The absorbance value in 2mL methanol is added for 2mLDPPH solution;A1It is that 2mL methanol adds 2mL various concentration The absorbance value of the mixture of sample;A2For 2mL DPPH methanol solution add 2mL various concentration sample absorbance value.
Test results are shown in figure 3, it can be seen from the figure that all samples remove DPPH within the scope of test concentrations Ability is in apparent concentration dependent.Under same concentrations, the Scavenging activity ratio TL-3 of sulfated polysaccharides STL-3 is significantly increased.Value It obtains one to be mentioned that, activity of the STL-3 in 25 μ g/mL of low dosage is almost twice higher than TL-3, or even than positive controls (Vc) It is higher.The IC of TLH-3, STLH-3 and Vc50Value is 69.3 μ g/mL, 9.0 μ g/mL and 21.2 μ g/mL respectively.According to it has been reported that The removing DPPH ability of few polysaccharide and sulfated polysaccharides from edible mushroom can exceed that STL-3.
(2) superoxide anion (O is removed2 -) ability
Prepare various concentration (25 μ g/mL, 50 μ g/mL, 100 μ g/mL, 200 μ g/mL, 300 μ g/mL, 400 μ g/mL, 500 μ G/mL the aqueous solution of the T.lobayense Heim polysaccharide STL-3 of T.lobayense Heim polysaccharide TL-3 and sulphation modification).
Prepare the sample aqueous solution mixing of the Tris-HCl buffer (50mM, pH value 8.3) and 0.4mL of 0.5mL, room temperature Concussion shakes up at 25 DEG C, stands 20min, is added 0.1mL pyrogallol (3mmol), is sufficiently mixed;Every 30s measurement at 325nm Light absorption value continues 5min, measures final light absorption value.Sample is replaced to compare group with distilled water, Vc makees positive control progress Above-mentioned experimental implementation.Every group experimental setup three parallel, is averaged.
·O2 -Scavenging activity calculated according to following formula:
Wherein, Δ A1For the final light absorption value of the polysaccharide solution of each concentration, Δ A0For pair for replacing sample with distilled water According to the light absorption value of group.
Test results are shown in figure 4, it can be seen from the figure that the IC of TL-3, STL-3 and Vc50Value is 111.6 μ g/ respectively ML, 27.1 μ g/mL and 29.9 μ g/mL.STL-3 shows the removing energy more taller than positive control within the scope of test concentrations Power.
(3) to the scavenging effect of hydroxyl radical free radical (OH)
Accurate formulation various concentration (10 μ g/mL, 25 μ g/mL, 50 μ g/mL, 100 μ g/mL, 200 μ g/mL, 400 μ g/ respectively ML, 500 μ g/mL) T.lobayense Heim polysaccharide TL-3, sulphation modification T.lobayense Heim polysaccharide STL-3 and Vc sample aqueous solution, 6mM FeSO4Solution, the H of 2.4mM2O2Solution, the alcohol-water poplar acid solution of 6mM.
Respectively take 1mL FeSO4Solution, 1mL alcohol-water poplar acid solution are added 1mL sample solution, reaction mixture are being tried It shakes and mixes in pipe, stand 10min.Add 1mL H2O2Solution reaction 30min.Using Vc as positive control, with masking foil point It Bao Guo not positive control, TL-3 and STL-3 water bath with thermostatic control 30min under the conditions of 37 DEG C.React the 2,3- dihydroxy benzenes first generated Acid has light absorption value at 510nm.It replaces polysaccharide solution as blank group using distilled water, replaces salicylic acid as control using distilled water Group, every group of experiment set three groups it is parallel.
The Scavenging activity of OH is calculated according to following formula:
Wherein, A0For the light absorption value of blank group, A2For the light absorption value of various concentration polysaccharide sample, A1For the extinction of control group Value.
Test results are shown in figure 5, it can be seen from the figure that the OH Scavenging activity ratio TL-3 of STL-3 and Vc is strong very much. All samples have obvious concentration dependent to the scavenging capacity of OH.The IC of TL-3, STL-3 and Vc50Value is respectively 173.8 μ G/mL, 73.5 μ g/mL and 50.1 μ g/mL.It will be apparent that sulfated polysaccharide STL-3 removing OH ability ratio TL-3 is more effective.
(4) reducing power
Prepare various concentration (10 μ g/mL, 25 μ g/mL, 50 μ g/mL, 100 μ g/mL, 200 μ g/mL, 400 μ g/mL, 500 μ G/mL the aqueous solution of the T.lobayense Heim polysaccharide STL-3 of T.lobayense Heim polysaccharide TL-3, sulphation modification);Preparing 0.2mol/L pH is The K that mass concentration is 1% is added after taking 0.2mL in 6.6 PBS buffer solution3Fe(CN)6Solution 1.0mL adds the sample of 1mL, Mixture is placed in 50 DEG C of water-bath 20min.Then, trichloroacetic acid (TCA) solution that 1.0mL mass concentration is 10% is added, 10min is centrifuged under 3000 revs/min.1.5mL supernatant is taken, 1% iron chloride (FeCl of 1.5mL distilled water and 0.2mL is added3) Solution.Mixture is protected from light 30min at 25 DEG C.Light absorption value is detected at 700nm.Make positive control with ascorbic acid (Vc), Sample is replaced to make blank with distilled water.The reducing power of sample is calculated according to the following formula:
Wherein A0Indicate the blank group light absorption value of not sample, A2Indicate the light absorption value of sample, A1Indicate no K3Fe(CN)6 The light absorption value of solution.
Test results are shown in figure 6, it can be seen from the figure that all samples show a degree of reducing power.TL-3, The IC of STL-3 and Vc50Value is respectively 274.2 μ g/mL, 90.5 μ g/mL and 64.8 μ g/mL.The reducing power of STL-3 compares TL-3 It significantly improves, and close to Vc.
The anti tumor activity in vitro of 4 products obtained therefrom of embodiment is tested:
(1) cell culture
HELF cell, HeLa cell and MCF-7 cell are cultivated in DMEM high glycoform culture medium.In DMEM added with 100mL/L fetal calf serum, 100U/mL penicillin and 100 μ g/mL streptomysins, in 37 DEG C, 5%CO2Incubator wet atmosphere Middle incubation.
(2) MTT is detected
The T.lobayense Heim polysaccharide STL-3 of T.lobayense Heim polysaccharide TL-3 and sulphation modification is to HeLa cell, MCF-7 cell and HELF The detection method for cytotoxicity of gas-liquid of cell: the 10 of 0.1mL are separately added into 96 orifice plates5Cell/mL density HeLa cell, MCF- 7 cells and after HELF cell culture 24 hours, by 5-Fu, TL-3 and STL-3 of various concentration (100 μ g/mL, 200 μ g/mL, 300 μ g/mL, 400 μ g/mL, 500 μ g/mL, 600 μ g/mL, 700 μ g/mL;100 μ L) be added separately to each culture medium, 37 DEG C CO2It is incubated for 24 hours in incubator.MTT solution (5mg/mL, 20 μ L) is added in each hole, is incubated 4 hours again.Remove training Support the DMSO standing 10min that 100 μ L are added in base.Absorbance is detected with microtiter plate reader at 570nm.Tumour cell The inhibiting rate of HeLa and MCF-7 and the survival rate formula of normal cell HELF are as follows:
Wherein A and B is the mean light absorbency of negative control and test sample respectively.
Test result is as shown in Figure 7 and Figure 8, it can be seen from figure 7 that the concentration with sample increases, the inhibition of sample Ability is all in dose dependent.To the inhibition growth ability aspect of HeLa cell, polysaccharide STL-3 significantly has than TL-3 stronger Rejection ability.In 200 μ g/mL, STL-3 is 4 times high to the inhibiting rate ratio TL-3's of HeLa cell.In addition, STL-3 is also shown Have apparent cytotoxicity to MCF-7 cell, 400 μ g/mL, STL-3 inhibiting rate almost 5 times more than TL-3.STL-3 pairs The IC of HeLa cell50Value is 216.7 μ g/mL, to the IC of MCF-7 cell50Value is 233.9 μ g/mL.By with other edible mushrooms Polysaccharide is compared, and STL-3 has fairly good advantage in terms of having tumor cytotoxic activity.As can be seen from Figure 8, with polysaccharide culture Cell still maintains very high survival rate after 24 hours, both TL-3 and STL-3 was without toxic side effect.The cell for reviewing 5-Fu is deposited Vigor gradually decreases, and in the concentration of 10 μ g/mL, 5-Fu is only 50% or so to the survival rate of HELF cell.In 400 μ g/mL, Influence of the 5-Fu to cell viability is 5 times of STL-3 or more.

Claims (3)

1.一种兼具抗肿瘤作用的抗氧化剂,其特征在于:所述抗氧化剂为硫酸化修饰的金福菇多糖STL-3;1. An antioxidant with anti-tumor effect, characterized in that: the antioxidant is sulfated modified mushroom polysaccharide STL-3; 所述硫酸化修饰的金福菇多糖STL-3的分子量为8.12×103Da,硫酸根含量为13.60%,取代度为1.22;The molecular weight of the sulfated modified chrysanthemum polysaccharide STL-3 is 8.12×10 3 Da, the sulfate content is 13.60%, and the substitution degree is 1.22; 所述硫酸化修饰的金福菇多糖STL-3由鼠李糖、葡萄糖、半乳糖、葡萄糖醛酸、阿拉伯糖、甘露糖和半乳糖醛酸7种单糖组成。The sulfated modified chrysanthemum polysaccharide STL-3 is composed of seven kinds of monosaccharides: rhamnose, glucose, galactose, glucuronic acid, arabinose, mannose and galacturonic acid. 2.根据权利要求1所述的兼具抗肿瘤作用的抗氧化剂,其特征在于:所述硫酸化修饰的金福菇多糖STL-3是以金福菇多糖TL-3为原料,利用三氧化硫-吡啶法进行硫酸化修饰获得。2. The anti-oxidant with anti-tumor effect according to claim 1, characterized in that: the sulphated-modified Jinfu mushroom polysaccharide STL-3 is made of Jinfu mushroom polysaccharide TL-3, and the three-oxidized Sulfur-pyridine method for sulfuration modification to obtain. 3.一种权利要求1或2所述的兼具抗肿瘤作用的抗氧化剂的制备方法,其特征在于:以纯度不低于95%的金福菇多糖TL-3为原料,加入原料质量35~50倍的溶剂甲酰胺,25℃水浴30min,再按原料质量的1~2倍加入三氧化硫-吡啶,60~80℃油浴下磁力搅拌3h,最后以4mol/L的NaOH调pH至中性,透析,浓缩,醇沉过夜,沉淀经冷冻干燥得金福菇硫酸化多糖STL-3,即为兼具抗肿瘤作用的抗氧化剂。3. A preparation method of an antioxidant with anti-tumor effect as claimed in claim 1 or 2, characterized in that: taking Jinfu mushroom polysaccharide TL-3 with a purity of not less than 95% as raw material, adding raw material quality 35 ~50 times the solvent formamide, 25 ℃ water bath for 30min, then add sulfur trioxide-pyridine according to 1~2 times the raw material mass, stir magnetically in 60~80℃ oil bath for 3h, and finally adjust the pH with 4mol/L NaOH to Neutralization, dialysis, concentration, alcohol precipitation overnight, and the precipitation was freeze-dried to obtain the sulfated polysaccharide STL-3 of Jinfu mushroom, which is an antioxidant with anti-tumor effect.
CN201610573418.0A 2016-07-20 2016-07-20 Antioxidant with anti-tumor effect and preparation method thereof Active CN106109487B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610573418.0A CN106109487B (en) 2016-07-20 2016-07-20 Antioxidant with anti-tumor effect and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610573418.0A CN106109487B (en) 2016-07-20 2016-07-20 Antioxidant with anti-tumor effect and preparation method thereof

Publications (2)

Publication Number Publication Date
CN106109487A CN106109487A (en) 2016-11-16
CN106109487B true CN106109487B (en) 2018-12-28

Family

ID=57289939

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610573418.0A Active CN106109487B (en) 2016-07-20 2016-07-20 Antioxidant with anti-tumor effect and preparation method thereof

Country Status (1)

Country Link
CN (1) CN106109487B (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106967179A (en) * 2017-03-30 2017-07-21 温岭市锦华铁皮石斛有限公司 It is a kind of to have antitumor and antioxidation Dendrobium officinale polysaccharide concurrently

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102603908A (en) * 2012-01-18 2012-07-25 安徽大学 Preparation method of natural antioxidant tricholoma lobayense heim polysaccharide
CN102659955A (en) * 2012-04-05 2012-09-12 浙江大学 Novel polysaccharide sulfating process based on physical-chemical combination
CN105520953A (en) * 2011-07-25 2016-04-27 香港理工大学 Tiger milk mushroom polysaccharide functionalized nano-selenium hydrosol with anti-tumor activity and preparation method thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105520953A (en) * 2011-07-25 2016-04-27 香港理工大学 Tiger milk mushroom polysaccharide functionalized nano-selenium hydrosol with anti-tumor activity and preparation method thereof
CN102603908A (en) * 2012-01-18 2012-07-25 安徽大学 Preparation method of natural antioxidant tricholoma lobayense heim polysaccharide
CN102659955A (en) * 2012-04-05 2012-09-12 浙江大学 Novel polysaccharide sulfating process based on physical-chemical combination

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
多糖及其硫酸化衍生物的抗病毒及免疫增强活性研究进展;赛福丁.阿不拉等;《天然产物研究与开发》;20131231;第25卷;第572-575,529页 *
秀珍菇多糖的硫酸化及其生物活性研究;申进文等;《河南农业科学》;20141231;第43卷(第7期);第102-106页 *

Also Published As

Publication number Publication date
CN106109487A (en) 2016-11-16

Similar Documents

Publication Publication Date Title
Hu et al. Chemical characterization, antioxidant, immune-regulating and anticancer activities of a novel bioactive polysaccharide from Chenopodium quinoa seeds
Patel et al. Physicochemical characterization, antioxidant and anti-proliferative activities of a polysaccharide extracted from psyllium (P. ovata) leaves
Wang et al. Sulfated modification of polysaccharides: Synthesis, characterization and bioactivities
Wang et al. Synthesis, characterization and antioxidant activity of selenium modified polysaccharides from Hohenbuehelia serotina
Xu et al. Characteristics and bioactivities of different molecular weight polysaccharides from camellia seed cake
Chen et al. Characterization of physicochemical and biological properties of Schizophyllum commune polysaccharide extracted with different methods
Sriramulu et al. Agaricus bisporus mediated biosynthesis of copper nanoparticles and its biological effects: An in-vitro study
Wang et al. Ultrasound-assisted extraction and analysis of maidenhairtree polysaccharides
Hu et al. Construction and structure-activity mechanism of polysaccharide nano-selenium carrier
Li et al. Effect of extraction method on structure and antioxidant activity of Hohenbuehelia serotina polysaccharides
Thanh et al. Structure and cytotoxic activity of ulvan extracted from green seaweed Ulva lactuca
Gheda et al. In vitro anticancer activity of polysaccharide extracted from red alga Jania rubens against breast and colon cancer cell lines
Gharibzahedi et al. Current emerging trends in antitumor activities of polysaccharides extracted by microwave-and ultrasound-assisted methods
Ye et al. Construction, characterization, and bioactive evaluation of nano-selenium stabilized by green tea nano-aggregates
Deng et al. Structural characterization, modification and hepatoprotective effects of polysaccharide from Mori Fructus
Álvarez-Viñas et al. Influence of molecular weight on the properties of Sargassum muticum fucoidan
Alboofetileh et al. Ultrasound‐assisted extraction of sulfated polysaccharide from Nizamuddinia zanardinii: Process optimization, structural characterization, and biological properties
Yu et al. Immunomodulatory activities of sulfated Cyclocarya paliurus polysaccharides with different degrees of substitution on mouse spleen lymphocytes
Singh et al. Curcumin loaded chitin-glucan quercetin conjugate: synthesis, characterization, antioxidant, in vitro release study, and anticancer activity
Vanavil et al. Bioactive and thermostable sulphated polysaccharide from Sargassum swartzii with drug delivery applications
Zhang et al. Physicochemical and antioxidant properties of Lycium barbarum seed dreg polysaccharides prepared by continuous extraction
Jing et al. Structural characterization and biological activities of a novel polysaccharide from Glehnia littoralis and its application in preparation of nano-silver
Shi et al. Synthesis, characterization, and cytotoxicity analysis of selenium nanoparticles stabilized by Morchella sextelata polysaccharide
Song et al. Sulfated polysaccharides from Rhodiola sachalinensis reduce d-gal-induced oxidative stress in NIH 3T3 cells
CN106967179A (en) It is a kind of to have antitumor and antioxidation Dendrobium officinale polysaccharide concurrently

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant