CN105111294A - Chinese yam peptide and application thereof - Google Patents
Chinese yam peptide and application thereof Download PDFInfo
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- CN105111294A CN105111294A CN201510563850.7A CN201510563850A CN105111294A CN 105111294 A CN105111294 A CN 105111294A CN 201510563850 A CN201510563850 A CN 201510563850A CN 105111294 A CN105111294 A CN 105111294A
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- chinese yam
- diabetic
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- peptide
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Abstract
The invention relates to the field of medicines and in particular relates to a Chinese yam peptide capable of preventing or treating diabetes mellitus. The sequence of the peptide is GQHSFYQLIHQGRVIPCDFIGVVP. A preparation method of the peptide comprises the following steps: (1) decoction in an alkaline solution, (2) neutralization, (3) enzymolysis, (4) protein separation, (5) separation and purification through semipreparative RP-HPLC (reverse phase high-performance liquid chromatography) and (6) freeze-drying, wherein amylase and diastase are adopted for enzymolysis; and macroporous resin and sephadex are adopted for protein separation. The invention provides an application of the peptide to treatment of diabetes mellitus and complications of diabetes mellitus, wherein the complications of diabetes mellitus include diabetic nephropathy, diabetic hypertension, diabetic eye diseases and diabetic neuropathy. The Chinese yam peptide is extracted from Chinese yam, has good safety and can be used for effectively preventing or treating diabetes mellitus.
Description
Technical field:
The present invention relates to pharmaceutical field, be specifically related to the polypeptide extract for preventing or treat diabetes.
Background technology
The metabolic disease of diabetes to be one group with hyperglycemia be feature is a kind of syndromes of glucose, protein and the lipid metabolism disorders caused by the cellular metabolism effect defect of insufficient insulin or Regular Insulin.In recent years, the number of diabetic subject increases gradually, is divided into type i diabetes and type ii diabetes.Wherein the number of type ii diabetes patient accounts for the 85-90% of diabetes total number of persons, and the health and lives of patient in serious threat.If diabetes can not get good control, a lot of complication may be caused, such as: diabetic nephropathy, retinopathy, hypertension etc.
At present, Most patients helps blood sugar in control volume by insulin injection or orally-taken blood sugar reducing medicine, or regulates the method treatment diabetes such as insulin secretion.But these drug side effects are comparatively large, clinically can not life-time service, the clinical needs of diabetic subject can not be met.If the medicine of the safe and effective treatment diabetes can extracted from natural product, will bring glad tidings for numerous diabetic subjects.
Chinese yam both can make staple food, can do vegetables again, nutritious, was just regarded as inexpensive qi-restoratives good merchantable brand since ancient times, had the good reputation of " Root of Glabrousleaf Asiabell ".Its property of Chinese yam is sweet, flat, nontoxic, returns spleen, lung, kidney channel.Cure mainly insufficiency of the spleen diarrhoea, cough due to deficiency of the lung, diabetes quenched one's thirst, oliguria frequently, seminal emission, women be with down and dyspeptic chronic intestinal inflammations.Chinese yam and glossy ganoderma are arranged in pairs or groups and take, and have the effect preventing and treating diabetes.Chinese yam is the medicine in mountain, the medicine in food, has the feature of integration of drinking and medicinal herbs.Therefore, Chinese yam not only has very high pharmaceutical use, and safe, nontoxic.However, the compound that extracts from Chinese yam of unripe exploitation comes out, for prevention or treatment diabetes.
Summary of the invention:
The present invention seeks to the feature for existing hypoglycemic medicine, design a kind of Chinese yam polypeptide, can safety, effectively treat diabetes.
Technical scheme
A kind of Chinese yam polypeptide, its sequence is GQHSFYQLIHQGRVIPCDFIGVVP.The preparation method of described polypeptide, is characterized in that: step comprises, and (1) decocts in basic solution, and (2) neutralize, (3) enzymolysis, (4) protein separation, RP-HPLC separation and purification is prepared, (6) freeze-drying in (5) half.Described enzymolysis adopts amylase, diastatic enzyme enzymolysis.Described protein separation adopts macroporous resin, dextrane gel.The application of described polypeptide in treatment diabetes and diabetic complication.Described diabetic complication is diabetic nephropathy, diabetic hypertension, diabetes eye illness, diabetic neuropathic pathology.
Beneficial outcomes:
Chinese yam polypeptide in the present invention, be extract polypeptide from Chinese yam, security is good, and can effectively prevent or treat diabetes.Be more preferably the purposes in treatment diabetes and/or diabetes complicated disease drug.Described diabetes comprise type i diabetes and type ii diabetes; Described diabetic complication includes but not limited to: diabetic nephropathy, diabetic hypertension, diabetes eye illness, diabetic neuropathic pathology etc.
Embodiment
Embodiment 1
The preparation method of Chinese yam polypeptide
After Chinese yam being added buck decoction 1.5h, be cooled to room temperature; With 1% hydrochloric acid adjust pH to neutral, add 2% amylase enzymolysis 2h; Adopt macroporous resin according to molecular weight isolated protein, then with half preparation RP-HPLC separation and purification, after the product freeze-drying obtained, be Chinese yam polypeptide.The Chinese yam polypeptide that application aforesaid method extracts, through Mass Spectrometric Identification, sequence is YERQADEPETKVTKKI, and this sequence is brand-new sequence.The present invention also can be synthesized by the raw work in Shanghai.
Embodiment 2
The cell in vitro determination of activity of Chinese yam polypeptide
Adopt MTT colorimetry.By the insulinoma cell (INS-1 cell) of logarithmic growth, add in 96 well culture plates with 1.0 × 105, cultivate 24h, experimental port, positive drug control hole add the Experimental agents Chinese yam polypeptide of different concns respectively; Blank group adds the solvent of same volume.Five multiple holes are established in every hole, and cultivate 48h, every hole adds MTT, after effect 4h, add DMSO, hatch 30min, absorbance A value is measured, by formula growth and proliferation of cell rate=(experimental group light absorption value/control group light absorption value-1) × 100% at microplate reader 570nm place.The EC50 calculating Experimental agents is 109.35nmol/L.
Embodiment 3
Hypoglycemic experiment in the body of Chinese yam polypeptide
Kunming mice, the high sugar of high fat is fed to body weight 18 ~ 22g, male and female half and half, and before experiment, water 24h is can't help in fasting, abdominal cavity disposable injection streptozotocin (STZ) 50mg/kg (citrate buffer of 1% dissolves preparation).After 1 week, blood sugar (BS) is surveyed in mouse tail vein blood sampling, and BS is modeling success higher than 16mmol/L.After modeling, mouse is divided into 3 groups, often organizes 10, diabetic model group (DM group): the PBS damping fluid subcutaneous injection giving equivalent; Control group (Bay g 5421): 2.0mg/kg subcutaneous administrations (being dissolved in 0.1mol/LPBS damping fluid), one day twice, continuous 15d; Sample sets (Chinese yam polypeptide): same to control group; Separately get 10 normal mouses as blank group.After medication, 15d is in tail venous blood sampling Quick Measurement blood sugar.After 15d, in drug withdrawal after 1 week, fasting be can't help water and is carried out glucose tolerance test in 16 hours.By 25% glucose solution (2g/kg) gavage, after gavage, 0,120 minute tail venous blood sampling detects blood sugar.
Result: glucose transporter protein polypeptide has the effect (P<0.05, table 1) reducing diabetic mice blood sugar; Meanwhile, can the sugar tolerance (table 2) of diabetes-alleviating model mice.The effect of its polypeptide is suitable with the effect of control group Bay g 5421.
Table 1 Chinese yam polypeptide is to mouse hypoglycemic activity
* p<0.05, * * p<0.01 is compared with model group
Table 2 Chinese yam polypeptide is to glucose tolerance in mice experimental result
* p<0.05, * * p<0.01 is compared with model group
Conclusion: Chinese yam polypeptide has therapeutic action to diabetic mice.
SEQUENCELISTING
Pu Luoda bio tech ltd, <110> Suzhou
<120> Chinese yam polypeptide and application thereof
<130>
<160>1
<170>PatentInversion3.5
<210>1
<211>24
<212>PRT
<213> artificial sequence
<400>1
GlyGlnHisSerPheTyrGlnLeuIleHisGlnGlyArgValIlePro
151015
CysAspPheIleGlyValValPro
20
Claims (6)
1. a Chinese yam polypeptide, is characterized in that: its sequence is GQHSFYQLIHQGRVIPCDFIGVVP.
2. a preparation method for polypeptide described in claim 1, is characterized in that: step comprises, and (1) decocts in basic solution, and (2) neutralize, (3) enzymolysis, (4) protein separation, and RP-HPLC separation and purification is prepared, (6) freeze-drying in (5) half.
3. the preparation method of polypeptide according to claim 2, is characterized in that: described enzymolysis adopts amylase, diastatic enzyme enzymolysis.
4. the preparation method of polypeptide according to claim 2, is characterized in that: described protein separation adopts macroporous resin, dextrane gel.
5. the application of polypeptide as claimed in claim 1 in preparation treatment diabetes and diabetes complicated disease drug.
6. application according to claim 5, is characterized in that described diabetic complication is diabetic nephropathy, diabetic hypertension, diabetes eye illness, diabetic neuropathic pathology.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510563850.7A CN105111294A (en) | 2015-09-08 | 2015-09-08 | Chinese yam peptide and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510563850.7A CN105111294A (en) | 2015-09-08 | 2015-09-08 | Chinese yam peptide and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105111294A true CN105111294A (en) | 2015-12-02 |
Family
ID=54659440
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510563850.7A Pending CN105111294A (en) | 2015-09-08 | 2015-09-08 | Chinese yam peptide and application thereof |
Country Status (1)
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| CN (1) | CN105111294A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103283929A (en) * | 2013-06-04 | 2013-09-11 | 焦作市易生元酒业股份有限公司 | Preparation method of dioscorea opposita polypeptide powder |
| CN103352065A (en) * | 2013-07-12 | 2013-10-16 | 湖北工业大学 | Huai rhizoma dioscoreae antioxidation polypeptide and preparation method thereof |
-
2015
- 2015-09-08 CN CN201510563850.7A patent/CN105111294A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103283929A (en) * | 2013-06-04 | 2013-09-11 | 焦作市易生元酒业股份有限公司 | Preparation method of dioscorea opposita polypeptide powder |
| CN103352065A (en) * | 2013-07-12 | 2013-10-16 | 湖北工业大学 | Huai rhizoma dioscoreae antioxidation polypeptide and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 邵海: "山药糖蛋白的分离纯化研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
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| C10 | Entry into substantive examination | ||
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| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20151202 |