CN102478503A - Method for detecting action mechanism of ursolic acid in inhibiting AGEs (advanced glycation end products) - Google Patents
Method for detecting action mechanism of ursolic acid in inhibiting AGEs (advanced glycation end products) Download PDFInfo
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- CN102478503A CN102478503A CN2010105581741A CN201010558174A CN102478503A CN 102478503 A CN102478503 A CN 102478503A CN 2010105581741 A CN2010105581741 A CN 2010105581741A CN 201010558174 A CN201010558174 A CN 201010558174A CN 102478503 A CN102478503 A CN 102478503A
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Abstract
The invention discloses a method for detecting an action mechanism of ursolic acid in inhibiting AGEs (advanced glycation end products), which includes the steps: firstly, simulating a non-enzymatic glycation system of body proteins; secondly, detecting inhibition of the ursolic acid against generation of fructosamine; thirdly, detecting inhibition of the ursolic acid against generation of AGEs; fourthly, processing data, statistically comparing inhibition of the ursolic acid against generation of the fructosamine to inhibition of the ursolic acid against generation of AGEs, and obtaining the action mechanism of the ursolic acid in inhibiting AGEs. The method can be used for simply and effectively detecting the action mechanism of the ursolic acid in inhibiting AGEs.
Description
Technical field
The invention belongs to the pathology detection field, concrete relate to a kind of method that ursolic acid suppresses the mechanism of action of AGEs that detects.
Background technology
Lasting hyperglycemia state in the human body; Can cause the reaction of many structure function albumen and nucleic acid-protein and glucose generation nonenzymatic glycosylation; Form terminal glycosylation product (Advance Glycation End Products; AGEs), AGEs is except that can influencing these albumen normal configuration and function, and it also can bring into play multiple pathological effect through terminal glycosylation acceptor (RAGE); Proved already: AGEs is that (Diabetic Mellitus, DM) patient quickens to take place the one of the main reasons of vascular lesion to diabetes.Therefore, suppressing AGEs and form, is the important means of control diabetic angiopathy.
(Ursolic Acid UA) has antitumor, anti-diabetic, lipopenicillinase, various biological effect such as anti-oxidant to ursolic acid, and we have confirmed that UA has the improvement effect to the DM injury of blood vessel at the zoopery in early stage, but its mechanism of action it be unclear that.
Summary of the invention
The object of the present invention is to provide a kind of method that ursolic acid suppresses the mechanism of action of AGEs that detects, may further comprise the steps:
Step 1: analogue body internal protein non-enzymatic glycation system;
Step 2: detect the inhibiting effect that ursolic acid generates fructosamine;
Step 3: detect the inhibiting effect that ursolic acid generates AGEs;
Step 4: data processing, utilize statistics comparison ursolic acid to inhibiting effect and the inhibiting effect of ursolic acid that fructosamine generates to the AGEs generation, promptly know the mechanism of action of ursolic acid to the inhibition of AGEs.
The present invention can simply, effectively detect the mechanism of action of ursolic acid to the inhibition of AGEs.
Embodiment
A kind of method that detects the mechanism of action of ursolic acid inhibition AGEs may further comprise the steps:
Step 1: analogue body internal protein non-enzymatic glycation system;
With bovine serum albumin(BSA) (BSA) concentration, these 3 factors of concentration of glucose and incubation time, 4 levels of each factor are with L16 (45) orthogonal test reaction system screening top condition.
In PBS liquid, add BSA, making its quality final concentration is 1,2,2.5,5g/L; D-glucose quality final concentration is 5,10,25,50mM; The adding final concentration is that the EDTA of 10mM is anticorrosion, after the aseptic filtration of 0.2m miillpore filter, places 37 ℃ of incubators, hatches 20; 40,60,80d constitutes table 1 orthogonal test factor level table.
Table 1 orthogonal test factor level table
Hatch when arriving setting-up time, sampling is with the fluorescent value of fluorospectrophotometer L55 (PE company) measurement glycosylated albumin.Excitation wavelength 370nm, emission wavelength 440nm, sensitivity 2, wide 8-10mm is stitched in gain 5.
Step 2: detect the inhibiting effect that ursolic acid generates fructosamine;
With best bovine serum albumin(BSA) nonenzymatic glycosylation system parameter (BSA concentration is 5g/L, and d-glucose is 50mM) is benchmark, under the aseptic condition, adds UA (with a small amount of DMSO dissolving; Being made into the storage liquid of 120M with PBS liquid) final concentration is 10M, 20M, 40M also sets up aminoguanidine (20mM) positive controls; The model control group of not dosing, and PBS (not containing albumen and glucose) is blank, sets up 3 parallel holes for every group; After hatching for 1 week, get each each 20L of group reaction liquid, add 1mL NBT (0.025mM) test solution respectively; 37 ℃ of water-baths, the strict control reaction time is with 10% acetum stopped reaction.At the 530nm place, with the zeroing of PBS blank, the absorbance when assaying reaction 5min and 15min (OD) value.Be calculated as follows the inhibiting rate of each group.Inhibiting rate=[1-(administration OD15-administration OD5)/(contrast OD15-contrast D05)] * 100%.
Step 3: detect the inhibiting effect that ursolic acid generates AGEs;
Under the aseptic condition, preparation protein nonenzymatic glycosylation vitro reactions system (BSA concentration is 5g/L, and d-glucose is 50mM) adds UA; Make its final concentration be respectively 10M, 20M, 40M, and set up aminoguanidine (20mM) positive controls; The model control group of not dosing, and PBS (not containing albumen and d-glucose) blank group are set up 6 parallel holes for every group; In 37 ℃ of incubators, cultivate 40d, working sample fluorescent value, detected parameters step 2.
Step 4: data processing, utilize inhibiting effect that statistics comparison ursolic acid generates fructosamine and ursolic acid to the inhibiting effect that AGEs generates, promptly know the mechanism of action of ursolic acid to the AGEs inhibition.
The foregoing description just is to let the one of ordinary skilled in the art can understand content of the present invention and enforcement according to this in order technical conceive of the present invention and characteristics to be described, to be its objective is, can not limit protection scope of the present invention with this.The variation or the modification of every equivalence that the essence of content has been done according to the present invention all should be encompassed in protection scope of the present invention.
Claims (1)
1. one kind is detected the method that ursolic acid suppresses the mechanism of action of AGEs, it is characterized in that, may further comprise the steps:
Step 1, analogue body internal protein non-enzymatic glycation system;
Step 2 detects the inhibiting effect that ursolic acid generates fructosamine;
Step 3 detects the inhibiting effect that ursolic acid generates AGEs;
Step 4, data processing utilizes statistics comparison ursolic acid to inhibiting effect and the inhibiting effect of ursolic acid to the AGEs generation that fructosamine generates, promptly knows the mechanism of action of ursolic acid to the inhibition of AGEs.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2010105581741A CN102478503A (en) | 2010-11-25 | 2010-11-25 | Method for detecting action mechanism of ursolic acid in inhibiting AGEs (advanced glycation end products) |
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| CN2010105581741A CN102478503A (en) | 2010-11-25 | 2010-11-25 | Method for detecting action mechanism of ursolic acid in inhibiting AGEs (advanced glycation end products) |
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| CN102478503A true CN102478503A (en) | 2012-05-30 |
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| CN2010105581741A Pending CN102478503A (en) | 2010-11-25 | 2010-11-25 | Method for detecting action mechanism of ursolic acid in inhibiting AGEs (advanced glycation end products) |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103884787A (en) * | 2014-02-21 | 2014-06-25 | 华南理工大学 | Detection method of peptide-advanced glycation end product |
| CN109106733A (en) * | 2017-06-22 | 2019-01-01 | 陕西天奎生物医药科技有限公司 | A kind of composition for preventing and treating diabetes and/or vascular complication disease, drug or health care product and its application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101686954A (en) * | 2007-07-09 | 2010-03-31 | 巴斯夫美容护理法国公司 | The inhibition that AGE forms |
| CN101732323A (en) * | 2009-12-10 | 2010-06-16 | 武汉大学 | Application of low-dose ursolic acid as medicament for treating diabetic early nephropathy |
| WO2010124521A1 (en) * | 2009-05-01 | 2010-11-04 | 常州高新技术产业开发区三维工业技术研究所有限公司 | Derivatives of rhein, preparation methods and uses thereof |
-
2010
- 2010-11-25 CN CN2010105581741A patent/CN102478503A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101686954A (en) * | 2007-07-09 | 2010-03-31 | 巴斯夫美容护理法国公司 | The inhibition that AGE forms |
| WO2010124521A1 (en) * | 2009-05-01 | 2010-11-04 | 常州高新技术产业开发区三维工业技术研究所有限公司 | Derivatives of rhein, preparation methods and uses thereof |
| CN101732323A (en) * | 2009-12-10 | 2010-06-16 | 武汉大学 | Application of low-dose ursolic acid as medicament for treating diabetic early nephropathy |
Non-Patent Citations (1)
| Title |
|---|
| DAWEI GAO ET AL.: "Study of the extraction,purification and antidiabetic potential of ursolic acid from Cornus officinalis Sieb. et Zucc.", 《THERAPY》, vol. 5, no. 5, 31 December 2008 (2008-12-31), pages 697 - 705 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103884787A (en) * | 2014-02-21 | 2014-06-25 | 华南理工大学 | Detection method of peptide-advanced glycation end product |
| CN103884787B (en) * | 2014-02-21 | 2015-06-03 | 华南理工大学 | Detection method of peptide-advanced glycation end product |
| CN109106733A (en) * | 2017-06-22 | 2019-01-01 | 陕西天奎生物医药科技有限公司 | A kind of composition for preventing and treating diabetes and/or vascular complication disease, drug or health care product and its application |
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Application publication date: 20120530 |