CN113599422B - Composition with anti-sugar effect and application thereof - Google Patents

Composition with anti-sugar effect and application thereof Download PDF

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CN113599422B
CN113599422B CN202111041930.8A CN202111041930A CN113599422B CN 113599422 B CN113599422 B CN 113599422B CN 202111041930 A CN202111041930 A CN 202111041930A CN 113599422 B CN113599422 B CN 113599422B
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parts
powder
roxburgh rose
composition
sugar
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CN113599422A (en
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梁鑫淼
王超然
郭志谋
徐伟
郭秀洁
伍言娟
董海临
刘亿宁
李效农
薛兴亚
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Taizhou Guokehuawu Biomedical Technologies Co ltd
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    • A61K36/48Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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Abstract

The application discloses a composition with an anti-sugar effect, which comprises the following raw materials in parts by weight: 1-50 parts of apple polyphenol, 1-50 parts of pagodatree flower bud, 1-50 parts of tea polysaccharide, 0.5-30 parts of roxburgh rose, 1-50 parts of ginseng and 0.1-30 parts of prebiotics; the composition has obvious AGEs formation inhibition effect and extremely strong oxygen free radical scavenging capability, has wide application prospect in the fields of antioxidation, anti-aging and anti-diabetes in the future, and can be used for preparing medicines, foods and health care products for treating, delaying or improving AGEs related diseases.

Description

Composition with anti-sugar effect and application thereof
Technical Field
The application relates to the fields of medicines, health-care products and foods, in particular to a composition with an anti-sugar effect and application thereof.
Background
Glucose and fructose metabolism produce Methylglyoxal (MG), a highly reactive byproduct, which can react with free amino groups in macromolecules such as proteins to produce reversible or irreversible conjugates, i.e., advanced glycation end products (advanced glycosylation end products, AGEs). MG exists in various tissues, organs and blood in the human body, and can enter the human body through an external source (such as high MG food), and can also be generated by in vivo metabolic disorders (such as diabetes). Studies have shown that: 1) MG can react with circulating proteins and lipoproteins, resulting in protein dysfunction, e.g., glycosylated hemoglobin, modified lipoproteins causing dyslipidaemia and atherosclerosis; 2) Accumulation of MG in the extracellular matrix may lead to modification of collagen and other structural proteins, thickening and fibrosis of the extracellular matrix, such as skin aging, loss of elasticity of the vessel wall; 3) In cells, MG induces oxidative stress, accumulation of misfolded proteins, endoplasmic reticulum stress, genotoxicity (chemical modification of DNA), etc., and may damage the body's detoxification system, causing cell damage in humans. For example, MG causes insulin resistance and beta cell dysfunction, are early factors in the development of type ii diabetes, and form a vicious circle between glycosylation and hyperglycemia. 4) In addition, MG is also an effective oxidative stress inducer. Both oxidative stress and MG levels are increased in diabetics, while the antioxidant defenses are significantly reduced, which results in increased glycosylation and the formation of a malignant cycle of ROS/AGE.
In conclusion, MG is extremely easy to react with in-vivo proteins to produce AGEs, causes various cell dysfunction and oxidative stress increase of human bodies, and is one of main causes of diabetic complications such as retinopathy, nephropathy and neuropathy; but also in the occurrence of cardiovascular diseases and central nervous system diseases such as cerebrovascular diseases and dementia.
Currently, viable strategies to reduce AGEs accumulation and toxicity include: 1) Regulating endogenous enzyme GLO system (GLO 1 catalyzes MG reaction to D-lactic acid), increasing human body detoxification function, and reducing AGEs generation; 2) Inhibiting the generation of AGEs induced by MG; 3) Increasing the scavenging of oxygen free radical ROS, breaking the self-vicious cycle of ROS/AGE.
Although many synthetic and natural components have been shown to inhibit AGEs, there is still a long way to follow from an effective treatment regimen. Among them, aminoguanidine is one of the most studied AGEs inhibitors at present. The micromolecular nucleophilic hydrazine compound has oxidation resistance, can eliminate hydroxyl free radicals and cut off the crosslinking of AGEs, can reduce the content of cholesterol and triacylglycerol and reduce the accumulation of exogenous AGEs in the body, but aminoguanidine and the compound thereof have adverse reactions found in clinical experiments and cannot be applied in clinical practice, so the stage of clinical research is exited. Therefore, development of novel AGEs inhibitors without adverse reactions is urgent.
Disclosure of Invention
The application provides a composition with an anti-sugar effect and application thereof.
The application provides a composition with an anti-sugar effect, which comprises the following raw materials in parts by weight: 1-50 parts of apple polyphenol, 1-50 parts of pagodatree flower bud, 1-50 parts of tea polysaccharide, 0.5-30 parts of roxburgh rose, 1-50 parts of ginseng and 0.1-30 parts of prebiotics.
Preferably, the raw materials comprise, by weight, 5-25 parts of apple polyphenol, 10-40 parts of pagodatree flower bud, 1-20 parts of tea polysaccharide, 1-10 parts of roxburgh rose, 1-5 parts of ginseng and 10-30 parts of prebiotics.
Preferably, the raw materials comprise, by weight, 15 parts of apple polyphenol, 35 parts of pagodatree flower bud, 15 parts of tea polysaccharide, 5 parts of roxburgh rose, 5 parts of ginseng and 25 parts of prebiotics.
Preferably, the tea polysaccharide is tea polysaccharide powder with content of 10-40%.
Preferably, the tea polysaccharide is prepared by extracting subtropical old tea with weak alkaline water, adding ethanol for precipitation and separation, and finally drying to obtain tea polysaccharide powder with the content of 10-40%.
Preferably, the apple polyphenol is apple polyphenol powder with the content of 20% -30%.
Preferably, the flos Sophorae Immaturus is flos Sophorae Immaturus extract or flos Sophorae Immaturus powder or flos Sophorae Immaturus instant powder with rutin content of 10-20%.
Preferably, the ginseng is ginseng powder or ginseng extract with 45-50% of ginsenoside content.
Preferably, the roxburgh rose is roxburgh rose powder or roxburgh rose extract or roxburgh rose juice freeze-dried powder or roxburgh rose juice; the prebiotics are selected from one or more of fructo-oligosaccharide, xylo-oligosaccharide and galacto-oligosaccharide.
Also provides the application of the composition with the anti-sugar effect in preparing medicines or foods for treating, delaying or relieving AGEs related diseases and health care products.
The application has the beneficial effects that:
1. the sugar-resistant composition has obvious AGEs formation-resistant activity and oxygen free radical scavenging activity and dose dependence through analysis of AGEs generation-resistant and oxygen free radical scavenging abilities;
2. the anti-sugar composition can obviously reduce the index of hyperglycemia such as fasting blood glucose, glycosylated hemoglobin, glycosylated serum protein, serum insulin, serum malondialdehyde and the like of a high-sugar diet mouse, and can obviously raise the level of superoxide dismutase in the serum of the high-sugar diet mouse. In conclusion, the anti-sugar composition can remarkably improve endogenous antioxidant capacity of the high-sugar diet mice, reduce the occurrence of saccharification of serum proteins and plasma proteins of the high-sugar diet mice, and reduce the blood sugar level of the high-sugar diet mice, so that the effects of improving and treating diabetes are achieved.
Drawings
FIG. 1 is a Trolox standard curve.
Detailed Description
The application is further described in connection with the following detailed description, in order to make the technical means, the creation characteristics, the achievement of the purpose and the effect of the application easy to understand.
The application provides the following technical scheme:
the composition with the anti-sugar effect comprises auxiliary materials, and the following components in parts by weight: 1-50 parts of apple polyphenol, 1-50 parts of pagodatree flower bud, 1-50 parts of tea polysaccharide, 0.5-30 parts of roxburgh rose, 1-50 parts of ginseng and 0.1-30 parts of prebiotics.
Preferably, the raw materials comprise, by weight, 5-25 parts of apple polyphenol, 10-40 parts of pagodatree flower bud, 1-20 parts of tea polysaccharide, 1-10 parts of roxburgh rose, 1-5 parts of ginseng and 10-30 parts of prebiotics.
Preferably, the raw materials comprise, by weight, 15 parts of apple polyphenol, 35 parts of pagodatree flower bud, 15 parts of tea polysaccharide, 5 parts of roxburgh rose, 5 parts of ginseng and 25 parts of prebiotics.
Preferably, the tea polysaccharide is specifically tea polysaccharide powder with the content of 10-40%.
Preferably, the tea polysaccharide is prepared by extracting subtropical old tea with weak alkaline water, adding ethanol for precipitation and separation, and finally drying to obtain tea polysaccharide powder with the content of 10-40%.
Preferably, the apple polyphenol is specifically apple polyphenol powder with the content of 20-30%.
Preferably, the flos Sophorae Immaturus is flos Sophorae Immaturus extract or flos Sophorae Immaturus powder or flos Sophorae Immaturus instant powder with rutin content of 10-20%.
Preferably, the ginseng is specifically ginseng powder or ginseng extract with 45-50% of ginsenoside content.
Preferably, the roxburgh rose is specifically roxburgh rose powder or roxburgh rose extract or roxburgh rose juice freeze-dried powder or roxburgh rose juice; the prebiotics are selected from one or more of fructo-oligosaccharide, xylo-oligosaccharide and galacto-oligosaccharide.
Preferably, the auxiliary material is selected from one or more of sweetener, sour agent, filler, lubricant, suspension, binder, excipient or preservative.
Also provides the application of the composition with the anti-sugar effect in preparing medicines or foods for treating, delaying or relieving AGEs related diseases and health care products.
Test example 1: AGE production inhibitor and oxygen radical ROS scavenger screening
1) AGE production inhibitor screening
Principle of: the non-enzymatic glycosylation reaction in living body refers to the process that aldehyde or ketone groups of reducing sugar react with free amino groups in macromolecules such as proteins to generate reversible or irreversible conjugate, namely advanced glycosylation end products (advanced glycation end products, AGEs) under the condition of no enzymatic catalysis. The protein glycosylation reaction model of bovine serum albumin-methylglyoxal (bovine serum albumin methylglyoxal, BSA-MGO) is adopted, the content of AGEs is measured by a fluorescence method, and the inhibition effect of each compound on the formation of AGEs is studied.
Test compound: apple polyphenol (20-30%), pagodatree flower bud powder (10-20%), tea polysaccharide (10-40%), rosa roxburghii powder and ginseng powder (45-50%) of ginsenoside.
Auxiliary materials: BSA (bi cloud), methylglyoxal MGO (sigma), aminoguanidine hydrochloride (MCE), 96-well clear master (Nunc).
The method comprises the following steps: the compound or aminoguanidine hydrochloride is dissolved in PBS, the concentration is 1mg/mL and 200 mu g/mL, 50 mu L of aminoguanidine hydrochloride (300 mu M) of positive medicine is respectively placed in a 96-well plate, 50 mu L of BSA solution and 10mg/mL of BSA solution are added, 250mM of MGO 50 mu L are sequentially added, incubation is carried out for 48 hours at 37 ℃, the mixture is placed in an enzyme-labeled instrument (lambda exc: 370 nm; lambda em: 440 nm) to detect fluorescence intensity, and the inhibition rate of AGEs formation of different compounds is calculated by substituting the following formula, and the result is shown in the table.
The calculation formula is as follows: AGEs inhibition (%) = (1-a sample/a blank) ×100%.
2) Oxygen radical scavenger screening
Principle of: the ability to scavenge oxidative free radicals is also known as antioxidant. The ORAC analysis method is based on the principle that the free radical breaks down the fluorescent probe to change the fluorescence intensity, and uses FLIPR to conduct real-time fluorescence analysis by taking the vitamin E water-soluble analogue Trolox as a quantitative standard. The magnitude of the change in fluorescence intensity reflects the degree of free radical destruction. In the presence of an antioxidant, it can inhibit the change in fluorescence caused by free radicals. The extent of inhibition reflects its antioxidant capacity towards free radicals.
Test compound: apple polyphenol (20-30%), pagodatree flower bud powder (10-20%), tea polysaccharide (10-40%), rosa roxburghii powder and ginseng powder (45-50%) of ginsenoside.
Auxiliary materials: a 96-well black transparent bottom plate is purchased from Nunc company; sodium fluorescein was purchased from Shanghai Source leaf company; AAPH (2, 2' -azobis (2-amidinopropane) dihydrochloride) available from apebrio corporation; PBS (phosphate buffer pH 7.4) was purchased from Gbico company; the detection platform is FLIPR-TETRA, the detected signal is relative fluorescence intensity (RFU), and the area under the fluorescence curve (AUC) is detected by utilizing the Screen works ™ system software in the detection platform.
The method comprises the following steps: the composition or Trolox was dissolved in PBS to prepare 1mg/mL and 200. Mu.g/mL concentrations, 50. Mu.L of each concentration was placed in a 96-well plate, and 100. Mu.L of 100nM sodium fluorescein was added. Incubation was carried out at 37℃for 10min, 100mM AAPH 50. Mu.L was added, and the fluorescence intensity was measured every 30s for a total of 1h. The area under the fluorescence curve AUC, net area = AUC sample-AUC phosphate buffer, and the antioxidant concentration, test agent concentration-net area standard curve was calculated. The Trolox standard curve is shown in fig. 1, and R2 is 0.9557. ORAC values were measured for the different compounds, respectively, and the results are shown in Table one.
3) Experimental results
Table 5 evaluation results of AGEs production inhibition and oxygen radical scavenging ability
Remarks: trolox standard curve as shown in FIG. 1
ORAC values are compared with reference to Trolox standard curve, and AGEs (R) inhibition rate is compared with reference to AGEs (R) inhibition rate of aminoguanidine hydrochloride, so that the results show that the above 5 samples can inhibit AGEs from generating and scavenging oxygen free radicals, and can be used for preventing or improving AGEs related diseases such as diabetes syndrome, atherosclerosis and the like. The medicine, food or health care product formed by the combination of the compounds can provide a new thought and approach for preventing the occurrence and development of AGEs related diseases.
Example 1
The composition related to the embodiment is prepared from the following raw materials in parts by weight:
the preparation method comprises the following steps: weighing and mixing the raw materials.
Example 2:
the composition related to the embodiment is prepared from the following raw materials in parts by weight:
the preparation method comprises the following steps: weighing and mixing the raw materials.
Example 3:
the composition related to the embodiment is prepared from the following raw materials in parts by weight:
the preparation method comprises the following steps: weighing and mixing the raw materials.
The AGEs inhibitory compositions obtained by mixing examples 1 to 3 were examined for their inhibitory effect on the formation of AGEs and their scavenging effect on oxygen radicals by referring to the method of test example 1.
The test results are as follows:
table two 3 results of evaluation of AGEs production inhibition and oxygen radical scavenging ability
ORAC values are compared with reference to Trolox standard curve, AGEs (R) inhibition rate is compared with reference to AGEs (R) inhibition rate of 1mg/ml aminoguanidine hydrochloride, and test results show that examples 1-3 have obvious AGEs formation resistance and oxygen free radical scavenging activity and have dose dependence.
Evaluation of antidiabetic efficacy of the anti-sugar composition on laboratory animals
Materials: example 2 composition (apple polyphenol 15 parts, locust flour 35 parts, tea polysaccharide 15 parts, roxburgh rose powder 5 parts, ginseng powder 5 parts, fructo-oligosaccharides 25 parts) db/db mice (Jizhikang), ELISA kit (Biyun days), metformin (Zhongshanghai Guibao pharmaceutical Co., ltd.).
The mice are adapted to the environment for 3 days, and fed with high-sugar high-fat mice after 3 days, the stomach-resistant composition (300 mg/kg at low dose and 500mg/kg at high dose) is infused regularly every day, the model control group is infused with normal saline, the positive drug group is metformin (300 mg/kg/d), and the continuous administration is carried out for 4 weeks. Whole blood is collected, and after standing, the whole blood is centrifuged, serum is separated, and blood glucose, glycosylated Serum Protein (GSP), glycosylated hemoglobin (HbA 1 c), INS (serum insulin), malondialdehyde content (MDA), superoxide dismutase activity (SOD) and the like in the blood are detected respectively by using a full-automatic biochemical analyzer and an ELISA method. The detection results are shown in tables three-eight, and the anti-sugar composition can obviously reduce the index of hyperglycemia such as fasting blood glucose, glycosylated hemoglobin, glycosylated serum protein, serum insulin, serum malondialdehyde and the like of the high-sugar diet mice, and can obviously raise the level of superoxide dismutase in the serum of the high-sugar diet mice. In conclusion, the anti-sugar composition can remarkably improve endogenous antioxidant capacity of the high-sugar diet mice, reduce the occurrence of saccharification of serum proteins and plasma proteins of the high-sugar diet mice, and reduce the blood sugar level of the high-sugar diet mice, so that the effects of improving and treating diabetes are achieved.
Table three: change of glycosylated hemoglobin (HbA 1 c)
Group of Animal number (only) HbA1c (%) was administered for 4 weeks
Model control group 10 14.2±1.11
Positive medicine group 10 9.71±1.51**
High dose group of anti-sugar compositions 10 9.03±1.14**
Low dose group of anti-sugar compositions 10 9.82±1.02**
Table four: changes in fasting blood glucose
Group of Animal number (only) Before administration (mmol/L) Administration was for 4 weeks (mmol/L)
Model control group 10 16.55±7.19 15.29±4.90
Positive medicine group 10 16.35±6.85 6.77±1.60**
High dose group of anti-sugar compositions 10 16.75±8.38 7.56±3.26**
Low dose group of anti-sugar compositions 10 16.34±5.43 9.77±2.22*
Table five: change of Glycosylated Serum Protein (GSP)
Group of Animal number (only) Administration is carried out for 4 weeks (umol/L)
Model control group 10 347.53±29.44
Positive medicine group 10 177.74±14.97**
High dose group of anti-sugar compositions 10 201.60±46.53**
Low dose group of anti-sugar compositions 10 251.00±13.61**
Table six: serum Insulin (INS) change
Group of Animal number (only) Administration is 4 weeks (mIU/L)
Model control group 10 55.66±5.32
Positive medicine group 10 27.43±3.38**
High dose group of anti-sugar compositions 10 27.31±1.99**
Low dose group of anti-sugar compositions 10 36.86±3.66**
Table seven: variation of superoxide dismutase (SOD) activity
Group of Animal number (only) Administration is 4 weeks (U/mL)
Model control group 10 161.73±22.22
Positive medicine group 10 301.64±33.01**
High dose group of anti-sugar compositions 10 279.41±14.32**
Low dose group of anti-sugar compositions 10 156.88±19.55*
Table eight: malondialdehyde content (MDA) variation
Group of Animal number (only) Administration was for 4 weeks (nmol/mL)
Model control group 10 15.88±2.29
Positive medicine group 10 7.27±2.62**
High dose group of anti-sugar compositions 10 8.05±2.35**
Low dose group of anti-sugar compositions 10 13.23±2.28*
The above examples merely illustrate specific embodiments of the application, which are described in more detail and are not to be construed as limiting the scope of the application. It should be noted that it is possible for a person skilled in the art to make several variants and modifications without departing from the technical idea of the application, which fall within the scope of protection of the application.

Claims (3)

1. The composition with the anti-sugar effect is characterized by comprising the following raw materials in parts by weight: 5-25 parts of apple polyphenol, 10-40 parts of pagodatree flower bud, 1-20 parts of tea polysaccharide, 1-10 parts of roxburgh rose, 1-5 parts of ginseng and 10-30 parts of prebiotics; the apple polyphenol is apple polyphenol powder with the content of 20-30%; the flos Sophorae Immaturus is flos Sophorae Immaturus extract or flos Sophorae Immaturus powder or flos Sophorae Immaturus instant powder with rutin content of 10-20%; the tea polysaccharide is prepared by extracting subtropical old tea with weak alkaline water, adding ethanol for precipitation separation, and finally drying to obtain tea polysaccharide powder with the content of 10-40%; the roxburgh rose is roxburgh rose powder or roxburgh rose extract or roxburgh rose juice freeze-dried powder or roxburgh rose juice; the Ginseng radix is Ginseng radix powder or Ginseng radix extract with ginsenoside content of 45-50%; the prebiotics are selected from one or more of fructo-oligosaccharide, xylo-oligosaccharide and galacto-oligosaccharide.
2. The composition with the anti-sugar effect according to claim 1, wherein the raw materials comprise, by weight, 15 parts of apple polyphenol, 35 parts of pagodatree flower bud, 15 parts of tea polysaccharide, 5 parts of roxburgh rose, 5 parts of ginseng and 25 parts of prebiotics.
3. Use of a composition with anti-glycaemic effect according to any of claims 1-2 for the preparation of a medicament for the treatment, delay or alleviation of diabetes.
CN202111041930.8A 2021-09-07 2021-09-07 Composition with anti-sugar effect and application thereof Active CN113599422B (en)

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