CN111184729A - Application of burdock inulin in preparation of medicine for treating hyperlipidemia - Google Patents
Application of burdock inulin in preparation of medicine for treating hyperlipidemia Download PDFInfo
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- CN111184729A CN111184729A CN202010098239.2A CN202010098239A CN111184729A CN 111184729 A CN111184729 A CN 111184729A CN 202010098239 A CN202010098239 A CN 202010098239A CN 111184729 A CN111184729 A CN 111184729A
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- 229920001202 Inulin Polymers 0.000 title claims abstract description 64
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 title claims abstract description 64
- 229940029339 inulin Drugs 0.000 title claims abstract description 64
- 235000003130 Arctium lappa Nutrition 0.000 title claims abstract description 55
- 235000008078 Arctium minus Nutrition 0.000 title claims abstract description 51
- 239000003814 drug Substances 0.000 title claims abstract description 16
- 208000031226 Hyperlipidaemia Diseases 0.000 title claims abstract description 7
- 238000002360 preparation method Methods 0.000 title claims description 6
- 240000005528 Arctium lappa Species 0.000 title abstract description 5
- 241000208843 Arctium Species 0.000 claims abstract description 50
- 201000005577 familial hyperlipidemia Diseases 0.000 claims abstract description 14
- 238000000502 dialysis Methods 0.000 claims abstract description 9
- 238000002386 leaching Methods 0.000 claims abstract description 9
- 230000003544 deproteinization Effects 0.000 claims abstract description 6
- 238000000746 purification Methods 0.000 claims abstract description 5
- 239000006228 supernatant Substances 0.000 claims description 17
- 238000005571 anion exchange chromatography Methods 0.000 claims description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 239000008367 deionised water Substances 0.000 claims description 12
- 229910021641 deionized water Inorganic materials 0.000 claims description 12
- 239000003480 eluent Substances 0.000 claims description 10
- 238000005227 gel permeation chromatography Methods 0.000 claims description 9
- 238000001914 filtration Methods 0.000 claims description 6
- 239000002244 precipitate Substances 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 3
- XOJVHLIYNSOZOO-SWOBOCGESA-N Arctiin Chemical compound C1=C(OC)C(OC)=CC=C1C[C@@H]1[C@@H](CC=2C=C(OC)C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)=CC=2)C(=O)OC1 XOJVHLIYNSOZOO-SWOBOCGESA-N 0.000 claims description 3
- 229920005654 Sephadex Polymers 0.000 claims description 3
- 239000012507 Sephadex™ Substances 0.000 claims description 3
- 229920002678 cellulose Polymers 0.000 claims description 3
- 239000001913 cellulose Substances 0.000 claims description 3
- 238000012869 ethanol precipitation Methods 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 3
- 239000003960 organic solvent Substances 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 239000011347 resin Substances 0.000 claims description 3
- 229920005989 resin Polymers 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 239000003463 adsorbent Substances 0.000 claims 1
- 108010028554 LDL Cholesterol Proteins 0.000 abstract description 15
- 108010023302 HDL Cholesterol Proteins 0.000 abstract description 14
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 abstract description 10
- 239000008280 blood Substances 0.000 abstract description 7
- 210000004369 blood Anatomy 0.000 abstract description 7
- 230000001603 reducing effect Effects 0.000 abstract description 6
- 238000011160 research Methods 0.000 abstract description 5
- 235000012000 cholesterol Nutrition 0.000 abstract description 4
- 230000001965 increasing effect Effects 0.000 abstract description 4
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 abstract description 4
- 238000010172 mouse model Methods 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 2
- 230000006698 induction Effects 0.000 abstract description 2
- 238000001556 precipitation Methods 0.000 abstract description 2
- 241000699670 Mus sp. Species 0.000 description 13
- 150000003626 triacylglycerols Chemical class 0.000 description 11
- 230000000694 effects Effects 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 239000003651 drinking water Substances 0.000 description 5
- 235000020188 drinking water Nutrition 0.000 description 5
- 239000013641 positive control Substances 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000037396 body weight Effects 0.000 description 3
- 229920002271 DEAE-Sepharose Polymers 0.000 description 2
- RYMZZMVNJRMUDD-UHFFFAOYSA-N SJ000286063 Natural products C12C(OC(=O)C(C)(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 RYMZZMVNJRMUDD-UHFFFAOYSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- RYMZZMVNJRMUDD-HGQWONQESA-N simvastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)C(C)(C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 RYMZZMVNJRMUDD-HGQWONQESA-N 0.000 description 2
- 229960002855 simvastatin Drugs 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 208000021959 Abnormal metabolism Diseases 0.000 description 1
- 241000208838 Asteraceae Species 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 238000011740 C57BL/6 mouse Methods 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
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- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 208000016097 disease of metabolism Diseases 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000006371 metabolic abnormality Effects 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Obesity (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Hematology (AREA)
- Diabetes (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to application of burdock inulin in preparing a medicament for treating hyperlipidemia. The burdock inulin is prepared by taking burdock roots as raw materials through leaching, alcohol precipitation, decoloration, deproteinization, dialysis and purification, and the weight average molecular weight of the burdock inulin is 500-1000 Da. The invention establishes a hyperlipemia mouse model through HFD (high fat) feed induction, researches the blood fat reducing activity of the Arctium lappa inulin BIP1-1, and finds that the Arctium lappa inulin BIP1-1 can play a certain role in treating hyperlipemia by reducing cholesterol (TC), Triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) level and increasing high-density lipoprotein cholesterol (HDL-C) level.
Description
Technical Field
The invention relates to application of burdock inulin in preparing a medicament for treating hyperlipemia, belonging to the technical field of medicines and health-care foods.
Background
Hyperlipidemia, which is a common metabolic disease in which elevated levels of cholesterol (TC), Triglycerides (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) are observed in serum, is caused by abnormal metabolism or transport of adipocytes in the human body. The existing research shows that hyperlipemia is closely related to diseases such as obesity, diabetes, atherosclerosis and the like, and is an important factor for causing cardiovascular and cerebrovascular diseases. Lipid-lowering therapy is one of effective means for preventing and treating the diseases, so that the development of an economic and effective lipid-lowering product has great significance.
Burdock (arctumlappa L.) is a biennial herb of compositae, and the existing research shows that the burdock has obvious functions of reducing blood sugar, blood pressure and weight, enhancing the immunologic function of human bodies, diffusing and removing heavy metals in water, and is widely used as a nutritional health-care raw material for food and medicines, but the specific bioactive component with the function of effectively reducing blood fat in the burdock is not reported yet.
Disclosure of Invention
The invention provides a new application of burdock inulin aiming at the defects of the research of the natural sugar medicine for treating hyperlipemia in the prior art, and in particular relates to the application of the burdock inulin in preparing the medicine for treating hyperlipemia.
The technical scheme of the invention is as follows:
the application of the burdock inulin in preparing the medicine for treating hyperlipemia is characterized in that the weight average molecular weight of the burdock inulin is 500-1000 Da.
The burdock inulin is prepared by taking burdock roots as raw materials and carrying out leaching, alcohol precipitation, decoloration, deproteinization, dialysis and purification.
According to the preferable preparation method of the burdock inulin, the steps are as follows:
(1) selecting fresh radix Arctii, peeling, slicing, leaching at 60-80 deg.C for 80-100min for 2-3 times, mixing the leaching solutions, filtering, vacuum filtering, and collecting supernatant;
(2) concentrating the supernatant obtained in the step (1) to 1/4-3/4 of the original volume, collecting the concentrated solution, performing ethanol precipitation, and keeping the precipitate; dissolving the precipitate in deionized water completely, centrifuging and collecting supernatant;
(3) carrying out decoloration and deproteinization treatment on the supernatant prepared in the step (2), removing residual organic solvent and retaining the solution;
(4) dialyzing the solution prepared in the step (3) at 4 ℃ by using a dialysis bag with the molecular weight cutoff of 500-1000Da to obtain a crude inulin solution BIP;
(5) fixing the volume of the crude inulin solution BIP obtained in the step (4) with deionized water until the crude inulin concentration is 10-15mg/mL, purifying by adopting an anion exchange chromatography column at the flow rate of 400-;
(6) performing primary separation and purification on the inulin solution obtained in the step (5) through an anion exchange chromatography column, and obtaining primarily purified burdock inulin BIP-1 by using 0.02M Tris-HCl as an eluent at the flow rate of 0.4-0.6 mL/min;
(7) and (4) further separating and purifying the primarily purified burdock inulin BIP-1 obtained in the step (6) by adopting gel chromatography, collecting target components, and freeze-drying to obtain solid burdock inulin BIP1-1 with pure white powder.
Further preferably, the leaching conditions in step (1) are 70 ℃, 90min, leaching for 2 times.
Further preferably, the supernatant in step (2) is concentrated to 1/3 of the original volume; the centrifugation is 7000r/min for 10 min.
Further preferably, the decoloring in the step (3) adopts D101 adsorption macroporous resin; the deproteinization adopts a Sevege method.
Further preferably, the dialysis bag in step (4) has a molecular weight cut-off of 500 Da; the dialysis time is 48-72h, preferably 48 h.
Further preferably, the concentration of the crude inulin in the step (5) is 12 mg/mL.
Further preferably, the anion exchange chromatography column in the step (5) is a DEAE-52 cellulose anion exchange chromatography column, the purified eluent is deionized water, and the flow rate is 500 μ L/min.
Further preferably, the anion exchange chromatography column in the step (6) is a DEAE Sepharose fast flow anion exchange chromatography column; the flow rate was 0.5 mL/min.
Further preferably, the gel chromatography in step (7) is Sephadex G-50 gel chromatography, the eluent for the gel chromatography is deionized water, and the flow rate of the eluent is 0.4-0.6mL/min, preferably 0.5 mL/min.
The medicine for treating hyperlipemia is characterized by comprising effective dose of the burdock inulin with the weight-average molecular weight of 500-1000 Da.
Preferably, according to the invention, the medicament comprises pharmaceutically acceptable excipients.
Has the advantages that:
the invention provides a new application of burdock inulin, in particular to an application of the burdock inulin in preparing a medicament for treating hyperlipemia, and provides a new idea and a new method for treating hyperlipemia. The invention establishes a hyperlipemia mouse model through HFD (high fat) feed induction, researches the blood fat reducing activity of the Arctium lappa inulin BIP1-1, and finds that the Arctium lappa inulin BIP1-1 can play a certain role in treating hyperlipemia by reducing cholesterol (TC), Triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) level and increasing high-density lipoprotein cholesterol (HDL-C) level.
Detailed Description
The technical solutions of the present invention are further described below with reference to specific examples, and it should be understood that the specific examples are only used for illustrating the technical solutions of the present invention, and do not limit the scope of the present invention. The materials and reagents mentioned in the examples are, unless otherwise specified, all common commercial products.
Example 1: preparation of burdock inulin
The preparation method of the burdock inulin comprises the following steps:
(1) selecting fresh radix Arctii, peeling, slicing, leaching at 70 deg.C for 90min for 2 times, mixing the extractive solutions, filtering, vacuum filtering, and collecting supernatant;
(2) concentrating the supernatant prepared in the step (1) to 1/3 of the original volume, collecting the concentrated solution, performing ethanol precipitation, and keeping the precipitate; dissolving the precipitate in deionized water completely, centrifuging at 7000r/min for 10min, and collecting supernatant;
(3) decoloring the supernatant prepared in the step (2) by using D101 adsorption macroporous resin, deproteinizing by using a Sevege method, removing residual organic solvent by using a rotary evaporator, and keeping the solution;
(4) dialyzing the solution prepared in the step (3) for 48h at 4 ℃ by using a dialysis bag with the molecular weight cutoff of 500Da to obtain a crude inulin solution BIP;
(5) fixing the volume of the crude inulin solution BIP obtained in the step (4) with deionized water until the crude inulin concentration is 12mg/mL, controlling the flow rate to be 500 muL/min, purifying with 2 times of column volume of deionized water through a DEAE-52 cellulose anion exchange chromatography column, separating inulin from glycoprotein and protein in the crude inulin solution BIP, and collecting the inulin solution;
(6) carrying out primary separation and purification on the synanthrin solution obtained in the step (5) through a DEAE Sepharose fast flow anion exchange chromatography column (1.6 multiplied by 20cm), taking 0.02M Tris-HCl as an eluent and the flow rate of 0.5mL/min, collecting one tube after 10min, detecting the sugar content in the eluent by a phenol-sulfuric acid method, and collecting the component with the sugar content being the highest peak position to obtain the primarily purified burdock synanthrin BIP-1;
(7) and (3) further separating and purifying the primarily purified burdock inulin BIP-1 obtained in the step (6) by Sephadex G-50 gel chromatography, collecting an elution solution by using an automatic fraction collector by using deionized water as an elution solution, mixing the same components, and freeze-drying to obtain the solid burdock inulin BIP1-1 with the pure white powder.
Example 2: effect of Burdock inulin on HFD (high fat) feed-induced hyperlipidemia mouse weight
The experimental method comprises the following steps:
establishment of a hyperlipemia mouse model:
male C57BL/6 mice, weighing 18 + -2 g/mouse, were randomly divided into 6 groups of 10 mice each after being acclimatized for one week;
blank group: feeding normal feed and common drinking water until the experiment period is finished;
model group: feeding high fat feed and common drinking water until the experiment period is finished;
positive control group: feeding high-fat feed, modeling for 12 weeks, and then performing intragastric gavage for 4 weeks by simvastatin (20 mg/kg/d);
burdock inulin low dose group (BIP1-1-L group): feeding high-fat feed and common drinking water, modeling for 12 weeks, and intragastrically administering Burdock inulin BIP1-1(10mg/kg/d) for 4 weeks;
burdock inulin middle dose group (BIP1-1-M group): feeding high-fat feed and common drinking water, modeling for 12 weeks, and intragastrically administering Burdock inulin BIP1-1(30mg/kg/d) for 4 weeks;
burdock inulin high dose group (BIP1-1-H group): feeding high-fat feed and common drinking water, modeling for 12 weeks, and intragastrically administering Burdock inulin BIP1-1(50mg/kg/d) for 4 weeks;
mice were weighed and recorded weekly.
Results analysis, the effect of Burdock inulin on the body weight of mice is shown in Table 1, and after 12 weeks of HFD feed feeding, the body weight of the mice in the model group is remarkably increased compared with that in the normal group (###P is less than 0.001), and after the gastric lavage treatment is carried out by adopting simvastatin and different doses of burdock inulin BIP1-1, the weight of mice in a positive control group, a BIP1-1-L group, a BIP1-1-M group and a BIP1-1-H group in a contrast model group are all obviously reduced in the sixteenth week (the weight of the mice in the positive control group, the BIP1-1-L group, the BIP1-1-M group and the BIP1-*P＜0.05，**P＜0.01，***P is less than 0.001). Therefore, the burdock inulin BIP1-1 has obvious inhibition effect on weight gain caused by HFD, thereby relieving the accumulation of blood fat to a certain extent.
Table 1. effect of burdock inulin BIP1-1 on mouse body weight (n ═ 10, x ± s)
Note: compared with the blank control group, the composition of the composition,###p is less than 0.001; in comparison to the set of models,*P＜0.05，**P＜0.01，***P＜0.001。
example 3: effect of Burdock inulin on mouse Cholesterol (TC), Triglycerides (TG), Low Density lipoprotein Cholesterol (LDL-C) and high Density lipoprotein Cholesterol (HDL-C)
The experimental method comprises the following steps: after the experimental period (16 weeks) of 6 groups of mice in example 2 was completed, the mice were fasted and water was not allowed to be supplied for 12 hours, blood was taken, the mice were left to stand at 4 ℃ for 2 hours and then centrifuged (3000rpm/min, 15min), the supernatant was aspirated and then centrifuged for the second time, and when the color of the supernatant was clear and free from impurities, the supernatant was aspirated into a sterile centrifuge tube and stored at-80 ℃. Then, the determination is carried out according to the instructions of the kit TC, TG and HDL-C, LDL-C of Nanjing construction company.
And (4) analyzing results: the effects of Burdock inulin on mouse cholesterol (TC), Triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) are shown in Table 2, and the levels of TC, TG and LDL-C in mice in a model group are remarkably improved compared with those in a normal group: (##P < 0.01), HDL-C levels are significantly reduced (##P is less than 0.01), and the TC, TG and LDL-C levels of mice in a positive control group, a BIP1-1-L group, a BIP1-1-M group and a BIP1-1-H group are obviously reduced compared with the model group (the level of TC, TG and LDL-C in the mice in the positive control group, the BIP1-1-L group and the BIP1-1-M group are obviously*P＜0.05，**P＜0.01，***P < 0.001), HDL-C levels significantly increased (*P＜0.05，**P is less than 0.01). Therefore, the burdock inulin BIP1-1 can obviously improve the abnormal blood lipid level caused by HFD, thereby prompting that the burdock inulin BIP1-1 has a certain effect of treating hyperlipidemia.
TABLE 2 Effect of Burdock inulin BIP1-1 on TC, TG and HDL-C, LDL-C in mice (n 10, x. + -. s)
Note: compared with the blank control group, the composition of the composition,##p is less than 0.01; in comparison to the set of models,*P＜0.05，**P＜0.01，***P＜0.001。
Claims (10)
1. the application of the burdock inulin in preparing the medicine for treating hyperlipemia is characterized in that the weight average molecular weight of the burdock inulin is 500-1000 Da.
2. The use according to claim 1, wherein the preparation method of the burdock inulin comprises the following steps:
(1) selecting fresh radix Arctii, peeling, slicing, leaching at 60-80 deg.C for 80-100min for 2-3 times, mixing the leaching solutions, filtering, vacuum filtering, and collecting supernatant;
(2) concentrating the supernatant obtained in the step (1) to 1/4-3/4 of the original volume, collecting the concentrated solution, performing ethanol precipitation, and keeping the precipitate; dissolving the precipitate in deionized water completely, centrifuging and collecting supernatant;
(3) carrying out decoloration and deproteinization treatment on the supernatant prepared in the step (2), removing residual organic solvent and retaining the solution;
(4) dialyzing the solution prepared in the step (3) at 4 ℃ by using a dialysis bag with the molecular weight cutoff of 500-1000Da to obtain a crude inulin solution BIP;
(5) fixing the volume of the crude inulin solution BIP obtained in the step (4) with deionized water until the crude inulin concentration is 10-15mg/mL, purifying by adopting an anion exchange chromatography column at the flow rate of 400-;
(6) performing primary separation and purification on the inulin solution obtained in the step (5) through an anion exchange chromatography column, and obtaining primarily purified burdock inulin BIP-1 by using 0.02M Tris-HCl as an eluent and at the flow rate of 0.4-0.6 mL/min;
(7) and (4) further separating and purifying the primarily purified burdock inulin BIP-1 obtained in the step (6) by adopting gel chromatography, collecting target components, and freeze-drying to obtain solid burdock inulin BIP1-1 with pure white powder.
3. The use of claim 2, wherein in step (2) the supernatant is concentrated to 1/3; the centrifugation is 7000r/min for 10 min.
4. The use of claim 2, wherein said decolorizing in step (3) employs D101 adsorbent macroporous resin; the deproteinization adopts a Sevege method.
5. The use of claim 2, wherein the dialysis bag in step (4) has a molecular weight cut-off of 500 Da; the dialysis time is 48-72 h.
6. The use of claim 2, wherein in step (5) the anion exchange chromatography column is a DEAE-52 cellulose anion exchange chromatography column and the purified eluent is deionized water.
7. The use of claim 2, wherein in step (6) said anion exchange chromatography column is a deaesephalose fast flow anion exchange chromatography column.
8. The use of claim 2, wherein the gel chromatography in step (7) is Sephadex G-50 gel chromatography, the eluent from the gel chromatography is deionized water, and the flow rate of the eluent is 0.4-0.6 mL/min.
9. A medicament for treating hyperlipidemia, which comprises an effective amount of the burdock inulin with the weight average molecular weight of 500-1000Da as claimed in claim 1.
10. The medicament of claim 9, wherein the medicament comprises a pharmaceutically acceptable excipient.
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| CN1485344A (en) * | 2003-09-02 | 2004-03-31 | 国家海洋局第一海洋研究所 | Burdock inulin and its preparing method |
| CN102965410A (en) * | 2012-12-07 | 2013-03-13 | 山东大学 | Method for extracting synanthrin from burdock |
| CN105037572A (en) * | 2015-07-03 | 2015-11-11 | 博德生物技术（北京）有限公司 | Method used for extracting burdock fructose oligosaccharides from burdock root |
| CN105106265A (en) * | 2015-06-25 | 2015-12-02 | 大连医科大学 | Application of burdok root oligosaccharide in preparation of antithrombotic drug |
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| CN1485344A (en) * | 2003-09-02 | 2004-03-31 | 国家海洋局第一海洋研究所 | Burdock inulin and its preparing method |
| CN102965410A (en) * | 2012-12-07 | 2013-03-13 | 山东大学 | Method for extracting synanthrin from burdock |
| CN105106265A (en) * | 2015-06-25 | 2015-12-02 | 大连医科大学 | Application of burdok root oligosaccharide in preparation of antithrombotic drug |
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