CN107898829A - Advanced glycation end products decomposition agent - Google Patents

Advanced glycation end products decomposition agent Download PDF

Info

Publication number
CN107898829A
CN107898829A CN201710879511.9A CN201710879511A CN107898829A CN 107898829 A CN107898829 A CN 107898829A CN 201710879511 A CN201710879511 A CN 201710879511A CN 107898829 A CN107898829 A CN 107898829A
Authority
CN
China
Prior art keywords
conditions
bsa
under
bhfs
age
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.)
Pending
Application number
CN201710879511.9A
Other languages
Chinese (zh)
Inventor
朴春红
霍越
李想
赵梓瀛
唐玉芳
刘俊梅
王玉华
于寒松
代伟长
周亚楠
王尚
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.)
Jilin Agricultural University
Original Assignee
Jilin Agricultural 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 Jilin Agricultural University filed Critical Jilin Agricultural University
Priority to CN201710879511.9A priority Critical patent/CN107898829A/en
Publication of CN107898829A publication Critical patent/CN107898829A/en
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Botany (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Biotechnology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Medical Informatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Nutrition Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

The invention discloses a kind of application of buckwheat shell chromocor extract in terms of advanced glycation end products decomposition agent is prepared, the buckwheat shell chromocor extract is to crush buckwheat shell, sieve, and adds water, ultra high pressure treatment；Filtering, is concentrated in vacuo, and freeze-drying, that is, obtain buckwheat shell chromocor extract；Solution is configured with phosphate buffer, as advanced glycation end products decomposition agent；Construct external non-enzymatic glycation system, the results showed that, buckwheat shell chromocor extract can crack advanced glycation end products.

Description

Advanced glycation end products decomposition agent
Technical field
The invention belongs to health food and medicine field, and in particular to advanced glycation end products decomposition agent.
Background technology
Advanced glycation end products（Advanced Glycation End Products, AGEs）Refer to reduced sugar and amino The whole latter stage product that acid or the macromolecular such as lipid are obtained through non-enzymatic glycation, building up for AGEs can cause in human body The generation and development of diabetes and its complication, a series of Alzheimer disease and angiocardiopathies.Numerous studies confirm, trigger The basic material of diabetic complication is AGEs.The source of AGEs is divided into endogenous and two kinds exogenous, by living organism What carbohydrate occurred saccharification reaction with protein and produced, it is referred to as endogenous AGEs, the formation of internal AGEs can raise internal Portugal Grape sugar utilization, aggravates insulin resistance and response to oxidative stress, causes intraor extracellular vascular endothelial cell by a variety of paths Change, disturb the normal function of the institutional framework of vitals；It is exogenous essentially from rich in carbohydrate and fatty More food, another reported smoking can produce AGEs, and the exogenous AGEs taken in from food, beverage or smoke from cigarette can be shown The concentration for raising it in blood is write, so as to trigger insulin resistance and accelerate tissue specificity to damage.Therefore with cut-out people AGE crosslinkings between body histone, the AGEs decomposition agents that decomposing should act on through the AGEs of formation become a variety of diseases for the treatment of New target drone.
1996, Vasan et al. first proposed AGEs decomposition agents this concepts, it is defined as that shape can be decomposed Into the material of AGEs, the AGE crosslinkings formed between AGEs histones can be cut off.The AGEs decomposition agents of first synthesis --- N- benzoyl thiazolium bromides are proved to reverse the AGE between diabetes collagen to be crosslinked, release and diabetes rat The crosslinked immunoglobulin G of erythrocyte surface（IgG）, and reverse the amyloid filaments modified formed by AGEs in vitro Aggregation.But in subsequent research, due to the unstability of the compound in aqueous, N- benzylidene thiazolium bromides Count out in the clinical practice stage.Then, Vasan proposed the compound of more stable, the active higher of structure in 2003 again ALT-711（4,5- dimethyl -3- phenylacetyl groups thiazole drone chlorides), through completing II phase clinical researches, similarly A series of AGEs decomposition agents also developed successively, such as C24, C36 and C36D2., can be anti-after the decomposition agent is combined with AGEs The structure of easy Spontaneous lysis should be generated, collagen is cut off and other biological macromolecular forms the bridge of cross-linked structure, make protein weight New separate out, and recover itself function therewith.ALT-711 can with reverting diabetes and hypertension animal model by AGE Cardiovascula arteriosclerosis caused by crosslinking.The research characteristic of AGEs decomposition agents is, is formed and stablized first under certain reaction system AGEs, then add decomposition agent and cracked, decomposition agent whole process is not involved in this process.In order to ensure the effect ring of decomposition agent Stablize interference-free, its requirement higher for non-enzymatic glycation system in border.Studied at present in existing AGEs decomposition agents In report, the research to reaction system influence factor is concentrated mainly on reactant, temperature with preparing the incubation time of AGEs and non- The several aspects of antibacterial mode of enzyme glycosylating system.
In the crops of northern China classics --- in buckwheat, contain abundant flavones.Buckwheat is polygonaceae （Polygonacca）Fagopyrum（Fagopyrum Mill）, it is distributed widely in Asia and Europe.Numerous studies evidence shows, buckwheat Containing flavone compounds such as substantial amounts of rutin, Quercetins in wheat, there is multiple pharmacological effect, such as prevention and treatment diabetes, Reducing blood lipid etc..Accessory substance of the buckwheat shell as buckwheat process, the technology famine in terms of higher value application, causes resource Significant wastage.Contain substantial amounts of Flavonoid substances in buckwheat shell, during the inhibitory action of its formation to AGEs is studied previous Sufficiently confirmed, but report is had no to the splitting action of AGEs.
The content of the invention
The object of the present invention is to provide a kind of Natively glycosylated endproduct cleavage agent.
Application of the buckwheat shell chromocor extract in terms of advanced glycation end products decomposition agent is prepared；
The buckwheat shell chromocor extract, is prepared by following methods：
1）Buckwheat shell is crushed, crosses 60-80 mesh sieves, by feed liquid mass ratio 1:5-20 adds water, hyperbaric heating；
2）Filtering, is concentrated in vacuo at 55-65 DEG C, is freeze-dried 20-30h, that is, obtains buckwheat shell chromocor extract.
Step 1）Described in hyperbaric heating be 121 DEG C at heat 15-25min.
Step 1）Described in hyperbaric heating be using ultra high pressure treatment device, pressure 100-400MPa, keep pressure 1- 5min, high-voltage-stable processing.
Step 1）Described in pressure be 300MPa, keep pressure time 2min.
The present invention provides application of the buckwheat shell chromocor extract in terms of advanced glycation end products decomposition agent is prepared.It is described Buckwheat shell chromocor extract be that buckwheat shell is crushed, is sieved, add water, ultra high pressure treatment；Filtering, is concentrated in vacuo, and freezing is dry It is dry, that is, obtain buckwheat shell chromocor extract；Solution is configured with phosphate buffer, as advanced glycation end products decomposition agent；Structure Build external non-enzymatic glycation system, the results showed that, buckwheat shell chromocor extract can crack advanced glycation end products.
Brief description of the drawings
Fig. 1 non-glycosylation system system fluorescent values, A are AGE-G-BSA systems, and B is AGE-F-BSA systems；
Fig. 2 BHFs crack AGE-G-BSA system fluorescent values；A is NaN3Under the conditions of concentration when being 0.05mg/mL；B is NaN3Bar When concentration is 0.20mg/mL under part；When concentration is 0.05mg/mL under the conditions of C is MSF；Concentration is 0.05mg/ under the conditions of D is MSF During mL；
Fig. 3 BHFs crack AGE-F-BSA system fluorescent values；A is NaN3Under the conditions of concentration when being 0.05mg/mL；B is NaN3Bar When concentration is 0.20mg/mL under part；When concentration is 0.05mg/mL under the conditions of C is MSF；Concentration is 0.05mg/ under the conditions of D is MSF During mL；
Fig. 4 non-glycosylation system system albumen average molecular weight；A is AGE-G-BSA systems NaN3Condition；B is AGE-G-BSA System MSF conditions；C is AGE-G-BSA systems NaN3Condition；D is AGE-G-BSA system MSF conditions；
AGE-G-BSA system glucose contents before and after Fig. 5 BHFs cracking；
AGE-F-BSA system AGEs fructose contents before and after Fig. 6 BHFs cracking；
AGE-G-BSA system AGEs free amine group contents before and after Fig. 7 BHFs cracking；
AGE-F-BSA system AGEs free amine group contents before and after Fig. 8 BHFs cracking.
Embodiment
The preparation of 1 advanced glycation end products decomposition agent of embodiment
Dried buckwheat shell is crushed with high speed disintegrator, crosses 60 mesh sieves, is put into clean beaker, in mass ratio 1:10 add Enter distilled water, under the conditions of 121 DEG C, hyperbaric heating 20min；Filtering, filter residue repeat extraction 2 times；Filtrate is merged, in 60 DEG C of bars Paste is concentrated in vacuo under part, freeze-drying 24h obtains buckwheat shell chromocor extract into powder（Buckwheat Hull Flavonoid extracts, BHFs）；By buckwheat shell chromocor extract phosphate buffer（PBS, 200mM, pH=7.4） The solution of 0.05mg/mL is configured to, as advanced glycation end products decomposition agent（BHFs）.
The preparation of 2 advanced glycation end products decomposition agent of embodiment
Dried buckwheat shell is crushed with high speed disintegrator, crosses 60 mesh sieves, is put into clean beaker, in mass ratio 1:10 add Enter distilled water, under the conditions of 121 DEG C, hyperbaric heating 20min；Filtering, filter residue repeat extraction 2 times；Filtrate is merged, in 60 DEG C of bars Paste is concentrated in vacuo under part, freeze-drying 24h obtains buckwheat shell chromocor extract into powder（Buckwheat Hull Flavonoid extracts, BHFs）；By buckwheat shell chromocor extract phosphate buffer（PBS, 200mM, pH=7.4） The solution of 0.2mg/mL is configured to, as advanced glycation end products decomposition agent（BHFs）.
The preparation of 3 advanced glycation end products decomposition agent of embodiment
Buckwheat shell is dried, is crushed with high speed disintegrator, 80 mesh sieves excessively, in mass ratio 1:20 add water, using ultra high pressure treatment Device carries out the high-voltage-stable processing of pressure 300MPa, pressurize 2min；Filtering, is concentrated in vacuo to paste under the conditions of 60 DEG C, cold Dry 24h is freezed into powder, that is, obtains buckwheat shell chromocor extract（Buckwheat Hull Flavonoid extracts, BHFs）；By buckwheat shell chromocor extract phosphate buffer（PBS, 200mM, pH=7.4）The solution of 0.2mg/mL is configured to, As advanced glycation end products decomposition agent（BHFs）.
The foundation of the external non-glycosylation system of embodiment 4
1. the foundation of external non-glycosylation system under the conditions of sodium azide
Respectively with being mixed with 0.02% sodium azide under the conditions of ultra-clean（NaN3）Phosphate buffer（PBS, 200mM, pH=7.4） Configure the bovine serum albumin(BSA) of 10.00mg/mL（Bovine Serum Albumin, BSA）The glucose of solution and 500mM （Glycation, G）Or fructose（Fructose, F）, into 5mL sugar juices add 10mL BSA solution, prepare respectively glycosylation- Glucose-bovine serum albumin(BSA) system（AGE-G-BSA）With glycosylation-fructose-bovine serum albumin(BSA) system（AGE-F-BSA）, Lucifuge is reacted 6 months under the conditions of 37 DEG C after mixing, takes out part sample at the 1st, 2,3,4,5,6 month respectively during this Product, are temporarily stored into -80 DEG C.
2. the foundation of micropore external non-glycosylation system under the conditions of filtering
By the way of being filtered on the basis of above-mentioned condition using miillpore filter (Micro-aperture suction filter, MSF) sterilize to reaction reagent.NaN is free of specifically, using3Phosphate buffer（PBS, 200mM, pH=7.4）Configuration is anti- Reagent is answered, reaction reagent is then passed sequentially through into the miillpore filter that aperture is 0.45 μm and 0.22 μm, after mixing at 37 DEG C Under the conditions of lucifuge react 6 months, took out sample segment at the 1st, 2,3,4,5,6 month respectively during this, be temporarily stored into -80 DEG C, then For AGEs to be cracked.
3. the cracking of external non-glycosylation system
It is 0.05mg/ to be separately added into concentration into the AGE-G-BSA systems and AGE-F-BSA systems prepared under the conditions of ultra-clean The glycosylation end products decomposition agent of mL and 0.20mg/mL, does not add the system of decomposition agent as blank group, the amino of same concentrations Guanidine（Aminoguanidine, AG）As positive control, lucifuge cracks 6 days under the conditions of 37 DEG C after mixing, Sample storage In -80 DEG C.
5 non-enzymatic glycation system dynamic experiment of embodiment
AGEs is a kind of complicated mixture, wherein contain the substantial amounts of material with fluorescence bridging property, such as pentosidine, thus it is glimmering Light value becomes the quantization important indicator of AGEs, the widely used method of researchers.Usually with sample in excitation wavelength The fluorescence intensity of 355nm, launch wavelength 460nm or so comes non-enzymatic glycation degree and AGEs growing amounts in performance system. The fluorescent value of cracking fore-and-aft architecture is measured at gain 50, excitation wavelength 355nm, launch wavelength 460nm using fluorescence microplate reader, And calculate cleavage rate using following equation：
（Formula 1）
As shown in Figure 1A, in AGE-G-BSA systems, in the reaction time in the time of 1-4 months, system fluorescent value is with reaction The growth of time and significantly increase（p<0.05）, when reaction proceeds to 5th month, fluorescent value significantly subtracted compared with 4th month It is few（p<0.05）, raised again when reaction proceeds to 6th month, be 135450.50 ± 50.00, be significantly higher than the 4th, the 5th month（p <0.05）,Reach more than its 3 at 1 month times.Fluorescent value in AGE-F-BSA systems is as shown in Figure 1B.In NaN3Condition Under, react the fluorescent value within 3 months and significantly increase（p<0.05）, but when reaction proceeds to 4th month, system fluorescent value It is significantly reduced to the trimestral 56.08%th（p<0.05）, following system fluorescent value the reaction of 4.5.6 months Significantly increase in time（p<0.05）；And under the conditions of MSF, system fluorescent value is in progressively growth trend substantially, up to 205647.50±731.00.The fluorescent value of AGE-F-BSA systems is significantly higher than AGE-G-BSA systems, this may with fructose with The reaction of BSA is relatively violent, and AGEs contents are more related in system.
Two kinds of different sterilization methods compare, the results showed that MSF group fluorescent values are substantially higher than NaN3Group, it is glimmering under the conditions of two kinds Light value is when reacting early stage（1st, 2 months）Difference and unobvious, start to show conspicuousness after reaction proceeds to 3rd month Difference（p<0.05）, especially in AGE-F-BSA systems when reaction proceeds to 4th month, MSF group fluorescent values are about NaN3Group 2.11 times.In order to simplify experimental procedure in ensuing experiment, the representative reaction time is selected as 1,3,6 month System the lytic effect of BHFs is studied.
The characteristic test of 6 advanced glycation end products decomposition agent BHFs of embodiment
1. effects of the BHFs to AGEs fluorescent values
（1）Effects of the BHFs to AGE-G-BSA system fluorescent values
AG is selected as positive control.Effects of the BHFs and AG to AGE-G-BSA system fluorescent values is as shown in Figure 2.As seen from the figure, In NaN3Under the conditions of, significant concentration-dose-dependence is presented in the inhibiting rate of BHFs（p<0.05）, its splitting action is significantly strong In AG（p<0.05）.Concentration be 0.05mg/mL under conditions of, BHFs to the reaction time be 3 months system splitting action most By force, and under conditions of concentration is 0.20mg/mL, BHFs is suitable to the system splitting action that the reaction time is 3,6 months, highest Reach 30.88 ± 0.04%（The system reaction time is 6 months）, the cleavage rate for being AG under the same terms（11.04±0.46%）'s 2.80 again.
Similar conclusion can also be obtained under the conditions of MSF.Under conditions of concentration is 0.20mg/mL, BHFs is to reaction The system splitting action highest of 6 months time, up to 27.85 ± 0.58%.Particularly, under the conditions of MSF, 0.05mg/mL BHF and AG to the reaction time be one month system inhibition it is weaker, especially the lytic effect of BHFs is only NaN3Bar 9.74% under part.Under conditions of concentration is 0.20mg/mL, the splitting action of BHFs and AG are weaker than NaN3Group.
（2）Effects of the BHFs to AGE-F-BSA system fluorescent values
As shown in figure 3, effects and AGE-G-BSA system of the BHFs to AGE-F-BSA system fluorescent values are different, the suppression of BHFs Significant concentration-dose-dependence is presented in rate processed（p<0.05）.From Fig. 3 A, Fig. 3 B, in NaN3Under the conditions of, in concentration Under conditions of 0.05mg/mL, the effect of BHFs and AG to system fluorescent value is relatively weak, and is 0.20mg/mL's in concentration Under the conditions of, the lytic effect of BHFs is significantly stronger than AG（p<0.05）, it is most strong to the system splitting action that the reaction time is 1 month, For 29.69 ± 0.67%.
Similar conclusion can also be obtained under the conditions of MSF, under conditions of concentration is 0.05mg/mL, BHFs is to reaction The system splitting action of 6 months time is significantly higher than AG（p<0.05）, when concentration is 0.20mg/mL, cracking of the AG to system Effect is obviously reduced with the growth in system reaction time（p<0.05）, the result with NaN3Under the conditions of result it is consistent, BHFs lytic effects are significantly stronger than AG（p<0.05）, and in NaN3Under the conditions of result unlike BHFs to the reaction time be 6 The system splitting action of a month is most strong, is 28.92 ± 0.57%, less than it in NaN3Under the conditions of highest cleavage rate.
2. effects of the BHFs to AGEs molecular weight of albumen
Pass through MALDI _ TOFMS（MALDI-TOF/TOF）Selection line in the positive-ion mode Property method to BHFs cracking fore-and-aft architecture molecular weight of albumen analyze, test generation initial data and collection of illustrative plates by 4000 SeriesExplorer V3.5 softwares export.
（1）Non-glycosylation system system molecular weight of albumen
MALDI-TOF/TOF is measured molecular weight analyte by the main peak in single system point spectrum, the increase of molecular weight It is significantly correlated with the extension of main peak, show to produce during non-enzymatic glycation it is a series of protein modified, can by Fig. 4 Know, the BSA molecular weight used in experiment is 66993.06Da, in blank group, with the growth in reaction time, system molecule Amount is stepped up, and AGE-G-BSA systems and AGE-F-BSA highests respectively reach 68758.74Da and 67740.39Da（It is MSF Condition）, 1765.68Da, 747.33Da are respectively increased compared with BSA.In contrast it is not difficult to find that under the same reaction conditions, The average molecular weight of AGE-G-BSA systems is more than AGE-F-BSA systems, and system molecular weight is consistently greater than under the conditions of MSF NaN3Condition.
（2）Effects of the BHFs to AGEs molecular weight of albumen
Average molecular weight results are shown in Table 1.System molecular weight is compared to control after adding the decomposition agent cracking that concentration is 0.20mg/mL The obvious increase of group, highest increases 209.06Da compared with blank group, is roughly equal to 4 carboxyls, in 24 systems after cracking, point Sample of the son amount amplification more than 45Da accounts for more than the 60% of sum.The result shows that in NaN3Under the conditions of, it is external after AG cracking Non-glycosylation system molecular weight is more than BHFs groups, and this result is completely contradicted under the conditions of MSF.And in AGE-G- In BSA systems, in NaN3Under the conditions of decomposition agent molecular weight increase degree be more than MSF conditions, especially the reaction time be 3 months System in, in NaN3Under the conditions of add AG and BHFs cracking after system molecular weight be respectively 67804.41Da and 67740.41Da, increases 209.06Da and 145.06Da respectively compared with blank group, and this amplification is only under the conditions of MSF 25.98Da and 39.62Da, i.e., in NaN3Under the conditions of amplification be respectively 8.04 and 3.66 times under the conditions of MSF.In other words, Though the blank molecular weight under the conditions of AGE-G-BSA systems MSF is more than NaN3Condition, but after its addition decomposition agent, the increasing of molecular weight Add degree smaller on the contrary.And this conclusion is in AGE-G-BSA systems and invalid.
AGEs albumen average molecular weight before and after 1 BHFs of table cracking
Note：Data are represented with the form of average value ± standard deviation in figure（N=3), same index difference lowercase alphabet registration According to, there are significant difference（p< 0.05）。
3. effects of the BHFs to AGEs content of reducing sugar
Nobio Glucose estimation kits are used respectively（Glucose oxidase-peroxidase method）And Sigma Frutose Glucose, the fructose content that Assay Kit crack system in fore-and-aft architecture are measured.
（1）Effects of the BHFs to AGE-G-BSA system AGEs glucose contents
Effects of the BHFs to glucose content in AGE-G-BSA systems is as shown in figure 5, in the blank group that the reaction time is 1 month In system, NaN3It is respectively 0.76 ± 0.02mg/mL and 0.55 ± 0.01mg/mL with glucose content under the conditions of MSF, is only anti- Answer initial concentration（30.00mg/mL）0.10%, show Maillard reaction at 1 month, the utilization of its glucose is basic Reach saturation state, in the system that the following reaction time is 3,6 months, glucose content is without significant changes（P ＞ 0.05）, and And under the conditions of MSF, system glucose content is less than NaN3Condition.In NaN3Under the conditions of, BHFs and AG are to system glucose content Effect it is not notable（P ＞ 0.05）, and in the system of MSF conditions, for the system that the reaction time is 3,6 months, BHFs Glucose content significantly raises after cracking（p<0.05）, it is respectively 0.62mg/mL ± 0.01 and 0.75 ± 0.01mg/mL, reaches 1.48 times of blank group and 1.80 times.
（2）Effects of the BHFs to AGE-F-BSA system AGEs fructose contents
It will be appreciated from fig. 6 that fructose content and glucose content variation tendency in AGE-G-BSA systems are basic in AGE-F-BSA systems Unanimously, in the system that the reaction time is 1 month, fructose content is in NaN3Be initial concentration 0.02% under the conditions of MSF；But Unlike glucose content variation tendency, in NaN3Under the conditions of, when reaction proceeds to 6th month, fructose contains in system Amount is remarkably decreased to 0.44 ± 0.02mg/mL（p<0.05）, it is only the 69.84% of the system that the reaction time is 3 months, and this As a result it is not notable under the conditions of MSF（P ＞ 0.05）.And to glucose content change is similar in AGE-G-BSA systems is, Fructose content under the conditions of AGE-F-BSA systems MSF is also below NaN3Group.The difference of fructose content and glucose content change it Place also resides in, and AG is in NaN3Obvious action is shown to the system that the reaction time is 3 months with the conditions of MSF（p< 0.05）, fructose content is significantly increased to 1.21 times and 1.14 times of the system of 1 month（p<0.05）, but BHFs is in NaN3Bar The part reaction time is but to significantly reduce fructose content in the system of 3 months（p<0.05）, it is only 0.38 ± 0.01mg/mL, and In the system that the reaction time is 6 months, with the addition of BHFs and AG, fructose content significantly raises（p<0.05）, especially In NaN3Under the conditions of system in, the fructose content after BHFs cracking is 1.80 times of blank group（p<0.05）.
4. effects of the BHFs to AGEs free amine group contents
Using the content of free amine group in OPA method measure systems.4.0mLOPA reagents are taken in test tube, add 200 μ L systems, it is right According to a group distilled water for addition same volume, after mixing, 2min is reacted under 35 DEG C of water bath conditions, is surveyed followed by microplate reader Determine light absorption value A of the system at 340nm340, with lysine replace sample using same procedure make standard curve, be y=- 0.1211x+3.5487(R2=0.998, OD value are in 0.18-0.90 scopes), according to the content of curve calculating free amino group.To rely Propylhomoserin concentration（X）(0.18~0.90mg/mL) is abscissa, the light absorption value A at 340nm340(Y) standard is drawn for ordinate Curve, equation of linear regression are y=- 0.1211x+3.5487, R2=0.998, free amine group content is in terms of percentage.
（1）Effects of the BHFs to AGE-G-BSA system AGEs free amine group contents
Free amine group in BSA molecular structures is active group necessary to fluorescence AGEs generations, thus free amine group content Change be often used to represent the degree of protein and sugared graft reaction.Fig. 7 shows, in AGE-G-BSA systems, with non-enzymatic The growth of glycosylation time, BSA free amine group contents after reduced sugar glycosylates substantially reduce（p<0.05）, when react into When row was by 6th month, system is in NaN3With free amine group content under the conditions of MSF be respectively 10.28 ± 0.01% and 6.32 ± 0.39%.Free aminoacid content is substantially less than NaN in the system of MSF conditions3Condition（p<0.05）.
The amino content that dissociates after BHFs and AG cracking in system significantly raises（p<0.05）, the two splitting action is suitable（p< 0.05）, and the two each system splitting action without significant difference（P ＞ 0.05）, MSF conditioned responses 6 after BHFs cracking The amino content that dissociates in the system of the moon reaches highest, is 11.56 ± 0.04%, and the free amine group under the same terms after AG cracking Content is 11.46 ± 0.05%, is respectively 1.82 times and 1.83 times of blank group.And this result is in NaN3Condition same reaction It is respectively 11.55 ± 0.01% and 11.50 ± 0.03% in the system of time.
（2）Effects of the BHFs to AGE-F-BSA system AGEs free amine group contents
As shown in Figure 8, the change of free amino content and AGE-G-BSA systems are different in AGE-F-BSA system blanks group, AGE-F-BSA systems are in the system that the reaction time is 1,3,6 month without significant changes（P ＞ 0.05）, 9.76% ~ 11.41% In the range of fluctuate, this result under the same conditions be higher than AGE-G-BSA systems in dissociate amino content.And and AGE-G-BSA Similarly, dissociate system amino content rise after addition decomposition agent in AGE-F-BSA systems, in NaN3In condition system, Although BHFs and AG be to the reaction time system of 1,3 month without remarkable effect, but make the system that the reaction time is 6 months In dissociate amino content significantly raise（p<0.05）, and the two effect is suitable, respectively 11.65 ± 0.02% and 11.63 ± 0.02%.In MSF condition systems, AG is significantly stronger than BHFs for the system splitting action that the reaction time is 1 month（p< 0.05）, it is that the amino content that dissociates after BHFs and AG cracking in system significantly raises in the reaction time in the system of 3,6 months, and And the two splitting action is suitable（p<0.05）.

Claims (5)

1. application of the buckwheat shell chromocor extract in terms of advanced glycation end products decomposition agent is prepared.
2. application according to claim 1, it is characterised in that：The buckwheat shell chromocor extract, is by following methods Prepare：
1）Buckwheat shell is crushed, crosses 60-80 mesh sieves, by feed liquid mass ratio 1:5-20 adds water, hyperbaric heating；
2）Filtering, is concentrated in vacuo at 55-65 DEG C, is freeze-dried 20-30h, that is, obtains buckwheat shell chromocor extract.
3. application according to claim 2, it is characterised in that：Step 1）Described in hyperbaric heating be 121 DEG C at heat 15-25min。
4. application according to claim 2, it is characterised in that：Step 1）Described in hyperbaric heating be using super-pressure at Device is managed, pressure 100-400MPa, keep pressure 1-5min, high-voltage-stable processing.
5. application according to claim 4, it is characterised in that：Step 1）Described in pressure be 300MPa, keep pressure Time 2min.
CN201710879511.9A 2017-09-26 2017-09-26 Advanced glycation end products decomposition agent Pending CN107898829A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710879511.9A CN107898829A (en) 2017-09-26 2017-09-26 Advanced glycation end products decomposition agent

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710879511.9A CN107898829A (en) 2017-09-26 2017-09-26 Advanced glycation end products decomposition agent

Publications (1)

Publication Number Publication Date
CN107898829A true CN107898829A (en) 2018-04-13

Family

ID=61840251

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710879511.9A Pending CN107898829A (en) 2017-09-26 2017-09-26 Advanced glycation end products decomposition agent

Country Status (1)

Country Link
CN (1) CN107898829A (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20030028234A (en) * 2001-09-27 2003-04-08 이형주 Extracts of buckwheat and/or buchwheat chaff with anti-inflammatory effects and composition comprising of the same
CN1634953A (en) * 2003-12-26 2005-07-06 山西省风陵渡开发区众力投资发展有限公司 Medicament for treating diabetes, diabetes complication, its preparation and novel use
CN103431389A (en) * 2013-08-27 2013-12-11 陕西科技大学 Method for continuously extracting buckwheat flavonoids and dietary fibres from buckwheat hulls
CN104825552A (en) * 2015-05-15 2015-08-12 吉林农业大学 Buckwheat shell flavonoid extract and application thereof as AGEs (advanced glycosylation endproducts) inhibitor

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20030028234A (en) * 2001-09-27 2003-04-08 이형주 Extracts of buckwheat and/or buchwheat chaff with anti-inflammatory effects and composition comprising of the same
CN1634953A (en) * 2003-12-26 2005-07-06 山西省风陵渡开发区众力投资发展有限公司 Medicament for treating diabetes, diabetes complication, its preparation and novel use
CN103431389A (en) * 2013-08-27 2013-12-11 陕西科技大学 Method for continuously extracting buckwheat flavonoids and dietary fibres from buckwheat hulls
CN104825552A (en) * 2015-05-15 2015-08-12 吉林农业大学 Buckwheat shell flavonoid extract and application thereof as AGEs (advanced glycosylation endproducts) inhibitor

Similar Documents

Publication Publication Date Title
Bhaskar et al. Beneficial effects of banana (Musa sp. var. elakki bale) flower and pseudostem on hyperglycemia and advanced glycation end-products (AGEs) in streptozotocin-induced diabetic rats
WO2016173545A1 (en) Broken ganoderma lucidum spore powder coffee and preparation method therefor
Takeuchi et al. Periodic high extracellular glucose enhances production of collagens III and IV by mesangial cells
DE69330767T2 (en) Process for the preparation of an amylase inhibitor
WO2021078296A1 (en) Polysaccharide-peptide complex for lowering blood sugar, blood lipid and glycosylated hemoglobin levels, and preparation method
Adewole et al. Effects of Ficus exasperata Vahl.(Moraceae) leaf aqueous extract on the renal function of streptozotocin-treated rats
JPH03240725A (en) Maillard reaction inhibitor
Rahmawati et al. Alpha-glucosidase inhibitory activity of stingless bee honey from Tetragonula biroi and Tetragonula laeviceps
JP5713571B2 (en) Maillard reaction inhibitor and AGEs production inhibitor
CN104825552A (en) Buckwheat shell flavonoid extract and application thereof as AGEs (advanced glycosylation endproducts) inhibitor
CN107898829A (en) Advanced glycation end products decomposition agent
JP7105523B2 (en) DIETARY FIBER AND METHOD FOR PRODUCING THE SAME
CN108567632A (en) Callus in Camellia sinensis extract is in the purposes of skin care
JP4477806B2 (en) Method for producing active ingredient of onion, active ingredient of onion by the production method and health food
Daugherty et al. Regional distribution of glycogen in the rat cestode, Hymenolepis diminuta
JP4233029B2 (en) Maitake extract, glycoprotein, and production method thereof
Koguchi et al. Dietary fiber suppresses elevation of uric acid and urea nitrogen concentrations in serum of rats with renal dysfunction induced by dietary adenine
CN107083294B (en) Solid-state fermentation black date wine and preparation method thereof
CN101884766A (en) Healthcare product or medicine for preventing diabetes complication and senium and preparation method thereof
AU2021101429A4 (en) Method for extracting flavonoids as active substances from honey
CN109419830A (en) Taiwan Chenopodium quinoa shell extract with whitening and antiaging effects, and its extraction and separation method
CN110522762A (en) Biological polysaccharide for preventing and treating inflammation and application thereof
CN112430516A (en) Anti-gout fermented strawberry wine and preparation method thereof
JPH08291192A (en) Lactonized oligosaccharide, its production and alpha amylase inhibitor
JP6565085B2 (en) Hypoglycemic symptoms relieving agent

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20180413