CN111743924A - Marine plant extract for treating diabetes, composition and application - Google Patents
Marine plant extract for treating diabetes, composition and application Download PDFInfo
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Abstract
The invention discloses a marine plant extract for treating diabetes, belonging to the technical field of medicinal extracts. The effective components include total polyphenol and/or total polysaccharide of carrageen. The invention also relates to a pharmaceutical composition of the marine plant extract and application thereof. Experiments show that the total polyphenol, polysaccharide and extract of carrageen have good antidiabetic effect and can inhibit the activity of diabetes target alpha-glucosidase. Provides a new candidate drug for treating diabetes, and further develops the medicinal value of the carrageen.
Description
Technical Field
The invention relates to a plant extract, in particular to a marine plant extract for treating diabetes.
Background
Diabetes is one of the main diseases with high morbidity and mortality at present, the number of patients with diabetes is remarkably increased at home and abroad, and insulin and oral hypoglycemic drugs are sequentially applied to clinic with the technological progress, however, the clinical application is limited due to the generation of various side effects. Therefore, the search for hypoglycemic drugs with less side effects and better curative effects is still a very concerned problem.
Regulating postprandial blood glucose levels is an effective therapeutic approach for the treatment of diabetes and the prevention of diabetes-related complications. Alpha-glucosidase is a very important enzyme in a living body, can be decomposed into monosaccharide by promoting carbohydrates such as starch, sucrose, maltose and the like in intestinal tract food, and enters blood circulation after being absorbed by epithelial cells on the upper segment of small intestine, so that postprandial blood sugar is increased, and therefore, the alpha-glucosidase has important clinical application value for treating and controlling diabetes.
Polysaccharides in food, such as starch, are digested into oligosaccharides with a small number of glucose molecules by saliva from the oral cavity and saliva from the pancreatic starch, and α -glucosidase cleaves α -1,4 glycosidic bonds from the non-reducing ends of these oligosaccharides to release glucose, which is absorbed by the small intestine epithelium and then enters the blood to become blood glucose.
Alpha-glucosidase inhibitors prevent the cleavage of oligosaccharides and disaccharides into monosaccharides by inhibiting the activity of alpha-glucosidase on the chorionic brush border of the small intestine epithelium, resulting in delayed glucose absorption. In addition, glucose absorption is further along the intestinal tract, thereby altering the release of gut hormones and enhancing insulin secretion. In general, alpha-glucosidase inhibitors lower postprandial insulin concentration levels by lowering postprandial blood glucose. The drug aiming at the target alpha-glucosidase is expected to become an effective hypoglycemic drug.
Ocean resources became a huge drug depot in the 21 st century. The particularity of the marine ecosystem, such as high pressure, high salt content, hypoxia, low illumination, etc., makes marine organisms have huge biological diversity and unique chemical diversity. Up to now, there are 17 drugs that are successfully marketed based on the development of marine natural products or their derivatives. Therefore, it is feasible to search and develop innovative drugs from marine products.
Chondrus ocellatus of the present inventionChondrus ocellatusHolmes) is an algae plant of the genus carrageen, also known as carrageen, gracilaria and gracilaria. Sweet taste and cold nature, and has the effects of clearing away heat and toxic materials and relieving constipation. Mainly treating swollen and sore throat, traumatic injury, cold and heat, mumps, epigastric pain, constipation due to intestinal dryness, etc.
Carrageen is an economically valuable alga. In addition to the manufacture of carrageenans, carrageenans are also edible and can be used as animal feed and agricultural fertilizer. At present, algae becomes an indispensable food in daily life of people, foreign nutriologists predict that the algae will become a popular 21 st century healthy food, and cloud-slice carrageen, calamus carrageen and the like are common dishes related to carrageen.
The carrageen is rich in nutrition, has a total sugar content of more than 50%, and also contains protein, phenolic substances, various amino acids and vitamins. At present, the algal polysaccharide has the functions of immunoregulation, tumor resistance, virus and bacteria resistance, ulcer resistance and the like. The polysaccharide has wide source and no cytotoxicity, and has small toxic and side effects when applied to organisms. Due to its unique chemical structure, algae polyphenol has various biological activities of bacteriostasis, antioxidation, anticancer, radioresistance, antianaphylaxis, antiasthmatic, antivirus, etc.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a novel marine plant extract aiming at the defects of the prior art, and the extract can be used for treating and preventing diabetes.
The technical problem to be solved by the present invention is achieved by the following technical means. The invention relates to a marine plant extract, which is characterized in that the extract is total polyphenol and/or total polysaccharide of carrageenin extracted from marine plant carrageenin.
The marine plant extract disclosed by the invention has the further preferable technical scheme that the extract is a mixture consisting of total carrageen polyphenols and total carrageen polysaccharides, and the weight ratio of the total carrageen polyphenols to the total carrageen polysaccharides in the mixture is 1: 10-10: 1.
The marine plant extract disclosed by the invention has the further preferable technical scheme that the extract is a mixture consisting of total carrageen polyphenols and total carrageen polysaccharides, and the weight ratio of the total carrageen polyphenols to the total carrageen polysaccharides in the mixture is 1: 3-3: 1.
The marine plant extract of the invention further preferably adopts the technical proposal that the extraction method of the total polyphenol and the total polysaccharide of the carrageen comprises the following steps:
(1) pulverizing and sieving carrageen, adding 2-20 times of 70-90% ethanol by volume, heating and extracting for 1-3 times in a dark condition, wherein the extraction time is 1-6 h each time, the extraction temperature is 40-70 ℃, combining filtrates, carrying out suction filtration, and carrying out freeze drying to obtain total carrageen polyphenol;
(2) taking filter residue obtained after extraction of the carrageenin, drying the filter residue at 40 ℃, adding 10-30 times of water, heating and extracting for 1-3 times, wherein the extraction time is 1-4 h each time, the extraction temperature is 70-100 ℃, combining the filtrate, carrying out rotary evaporation at 40-70 ℃ to a proper volume, adding 3-5 times of volume of 95% ethanol, and carrying out alcohol precipitation overnight; centrifuging to obtain precipitate, and freeze drying to obtain carrageenan total polysaccharide.
The marine plant extract can be used for preparing a medicament for treating diabetes.
The invention also discloses a pharmaceutical composition for inhibiting the activity of the alpha-glucosidase, which is characterized in that the pharmaceutical composition is prepared by taking marine plant extracts as main components and adding pharmaceutically acceptable pharmaceutic adjuvants. The dosage form of the medicament can be a conventional dosage form.
Compared with the prior art, the invention has the following beneficial effects:
the invention finds two components in the carrageen extractive which have the effect of preventing and treating diabetes, and the adopted extraction method is simple, convenient and practical, and provides a certain reference value for further industrial production. The medicine is prepared by utilizing the blood sugar reducing effect of the carrageenan and the polyphenol, so that the diabetes can be prevented and treated.
Detailed Description
The following further describes embodiments of the present invention to enable those skilled in the art to understand the present invention without limiting the claims of the present invention.
Example 1, a marine plant extract, which is total polyphenol of carrageen extracted from marine plant carrageen. The extraction method of total polyphenol of carrageen comprises the following steps: pulverizing and sieving carrageen, adding 10 times volume of 80% ethanol, heating and extracting for 2 times in dark condition, wherein the extraction time is 2h each time, the extraction temperature is 60 ℃, combining filtrates, filtering, and freeze-drying to obtain carrageen total polyphenol. The medicine is prepared by taking total carrageenin polyphenol as a main component and adding pharmaceutically acceptable medicinal auxiliary materials, and is used for treating diabetes.
Example 2, a marine plant extract which is the total polysaccharide of carrageenans extracted from the marine plant carrageenans. The extraction method of the total carrageenin polysaccharide comprises the following steps:
pulverizing and sieving carrageen, adding 15 times volume of 70% ethanol, heating and extracting for 3 times in dark condition, wherein the extraction time is 4h each time, the extraction temperature is 60 ℃, combining filtrates, filtering, and freeze-drying to obtain carrageen total polyphenol; taking filter residue after extracting carrageenan polyphenol, drying at 40 ℃, adding 20 times of water, heating and extracting for 2 times, wherein the extraction time is 2 hours each time, the extraction temperature is 80 ℃, combining filtrates, carrying out rotary evaporation at 50 ℃ to a proper volume, adding 3 times of volume of 95% ethanol, and carrying out alcohol precipitation overnight; centrifuging to obtain precipitate, and freeze drying to obtain carrageenan total polysaccharide.
The medicament is prepared by taking the total polysaccharide of the carrageenin as a main component and adding pharmaceutically acceptable pharmaceutic adjuvants. Can be used for treating diabetes.
Example 3, a marine plant extract which is total polyphenol and total polysaccharide of carrageen extracted from marine plant carrageen. The weight ratio of total carrageenin polyphenols to total carrageenin polysaccharides is 1: 1.
The extraction method of the total carrageenin polyphenol and the total carrageenin polysaccharide comprises the following steps:
(1) pulverizing and sieving carrageen, adding 90% ethanol 5 times the volume of the carrageen, heating and extracting for 3 times in the dark condition, wherein the extraction time is 2h each time, the extraction temperature is 70 ℃, combining the filtrates, filtering, and freeze-drying to obtain total carrageen polyphenol;
(2) taking filter residue after extracting carrageenan polyphenol, drying at 40 ℃, adding 10 times of water, heating and extracting for 2 times, wherein the extraction time is 3h each time, the extraction temperature is 85 ℃, combining filtrates, carrying out rotary evaporation at 55 ℃ to a proper volume, adding 4 times of volume of 95% ethanol, and carrying out alcohol precipitation overnight; centrifuging to obtain precipitate, and freeze drying to obtain carrageenan total polysaccharide.
The medicament is prepared by taking total carrageenin polyphenol and total carrageenin polysaccharide as main components and adding pharmaceutically acceptable pharmaceutic adjuvants. Can be used for treating diabetes.
Example 4, a marine plant extract experiment one:
pulverizing and sieving carrageen, weighing 1000g of carrageen powder, extracting for 2 times (500 g each time), adding 80% ethanol 15 times the volume of each time, heating and extracting for 3.5h at 50 deg.C in dark condition, repeatedly extracting for 3 times, mixing filtrates, vacuum filtering, and freeze drying to obtain total polyphenol of carrageen.
Taking the residue after extracting carrageenan polyphenol, drying at 40 deg.C, extracting for 2 times, adding 20 times of water each time, heating and extracting at 90 deg.C for 2h, repeating extracting for 2 times, mixing filtrates, rotary evaporating at 60 deg.C to appropriate volume, adding 4 times of 95% ethanol, precipitating with ethanol overnight, centrifuging, and collecting precipitate to obtain crude polysaccharide.
Inhibition of alpha-glucosidase by Carrageenan extract:
the screening model establishes a reaction system by taking PNPG as a substrate. Adding 50 muL of sample solution, and setting 3 samples per group per concentrationIn parallel, additionally arranging a sample control, a blank reaction and a blank control, adding 50 mu L of 0.45U/mL enzyme solution and a PH6.8 PBS buffer solution, uniformly mixing, reacting in a water bath at 37 ℃ for 10min, then adding 100 mu L of PNPG (5 mmol/L), uniformly mixing, reacting in a water bath at 37 ℃ for 20min, and then adding 100 mu L of 0.2 mol/L Na2CO3Stopping reaction, measuring OD value at 405nm of microplate reader, using acarbose as positive control liquid medicine, measuring the influence of different concentration sample liquid on α -glucosidase inhibition activity, calculating inhibition rate, using Origin 8.0 software to make inhibition rate-concentration graph and make regression equation fitting to obtain half inhibition mass concentration (IC)50）。
The experimental results are as follows:
the results of enzyme activities of the sample group and the control group are shown in Table 1. Carrageenan polyphenol, carrageenan and a mixture of carrageenan polyphenol and polysaccharide (mass ratio 1: 1) all significantly inhibited the activity of alpha-glucosidase compared to the control group.
TABLE 1 Effect of Chondrus crispus extract and acarbose on alpha-glucosidase Activity
Example 5, a marine plant extract experiment two:
EXAMPLE 4 Effect of carrageen extract on type 2 diabetic mice
Experimental materials: male mouse, mouse feed, carrageenin, mixture of carrageenin and carrageenin (mass ratio 1: 1)
The experimental method comprises the following steps:
animal grouping: healthy mice were randomly divided into 9 groups, including a blank control group, a model group, a positive control group (acarbose) and a carrageenan polyphenol high dose group, a carrageenan polyphenol low dose group, a carrageenan polysaccharide high dose group, a carrageenan polysaccharide low dose group, a mixture of carrageenan polyphenol and polysaccharide high dose group, a mixture of carrageenan polyphenol and polysaccharide low dose group, 10 mice per group.
Establishing a diabetes model: after the mice are purchased, the adaptive words are raised for 1 week, after the mice are fasted and water is not forbidden for 12 hours, 10 healthy mice are randomly selected and injected with normal saline in the abdominal cavity to serve as a blank control group; the rest healthy mice are injected with 50mg/kg of alloxan solution (prepared at present). After 72 hours, selecting the mice with the blood sugar value of more than or equal to 11.1 mmol/L as the mice successfully modeled.
Administration: the blank control group and the model group are respectively given with physiological saline; the positive control group is given acarbose tablet solution; the sample groups were given different doses of carrageenan extract suspension, 30, 60 mg/kg respectively. 4 weeks after administration, the mice in each group were recorded for fasting plasma glucose, glucose tolerance, and serum cholesterol TC and triglyceride TG levels.
Table 2 effect of carrageenan extract on fasting plasma glucose in tetrahydropyrimidine model mice ("X ± S,n=10, # # # P compared to model group<0.001）
The experimental results are as follows:
the results of the fasting blood glucose levels in the mice after 4 weeks of administration are shown in Table 2. After 4 weeks, fasting blood glucose levels were significantly lower in the blank animals than in the model group (P < 0.001), indicating successful modeling of the hyperglycemic model. Compared with the model group, the blood sugar of each administration group is obviously reduced (P is less than 0.001), which shows that each extract has obvious effect of reducing blood sugar.
Claims (6)
1. A marine plant extract is characterized in that the extract is total polyphenol and/or total polysaccharide of carrageen extracted from marine plant carrageen.
2. The marine plant extract as claimed in claim 1, wherein the extract is a mixture of total carrageen polyphenols and total carrageen polysaccharides, and the weight ratio of the total carrageen polyphenols to the total carrageen polysaccharides in the mixture is 1: 10-10: 1.
3. The marine plant extract as claimed in claim 1, wherein the extract is a mixture of total carrageen polyphenols and total carrageen polysaccharides, and the weight ratio of the total carrageen polyphenols to the total carrageen polysaccharides in the mixture is 1:3 to 3: 1.
4. The marine plant extract according to claim 1, 2 or 3, wherein said extraction method of total carrageenans polyphenols and total carrageenans polysaccharides is as follows:
pulverizing and sieving carrageen, adding 2-20 times of 70-90% ethanol by volume, heating and extracting for 1-3 times in a dark condition, wherein the extraction time is 1-6 h each time, the extraction temperature is 40-70 ℃, combining filtrates, carrying out suction filtration, and carrying out freeze drying to obtain total carrageen polyphenol;
(2) taking filter residue obtained after extraction of the carrageenin, drying the filter residue at 40 ℃, adding 10-30 times of water, heating and extracting for 1-3 times, wherein the extraction time is 1-4 h each time, the extraction temperature is 70-100 ℃, combining the filtrate, carrying out rotary evaporation at 40-70 ℃ to a proper volume, adding 3-5 times of volume of 95% ethanol, and carrying out alcohol precipitation overnight; centrifuging to obtain precipitate, and freeze drying to obtain carrageenan total polysaccharide.
5. The use of the marine plant extract as claimed in any one of claims 1 to 4, wherein the use is the use of the marine plant extract for the preparation of a medicament for the treatment of diabetes.
6. A pharmaceutical composition for inhibiting the activity of alpha-glucosidase, which is characterized in that the pharmaceutical composition is prepared by taking the marine plant extract as the main component according to any one of claims 1 to 4 and adding pharmaceutically acceptable pharmaceutical excipients.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114276471A (en) * | 2021-12-30 | 2022-04-05 | 江苏海洋大学 | Preparation method and application of caulerpa lentillifera polysaccharide |
| CN115813958A (en) * | 2022-11-17 | 2023-03-21 | 江苏海洋大学 | Application of carrageenin in preparing medicine for treating Alzheimer disease and preparation method thereof |
-
2020
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Non-Patent Citations (2)
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| 冯学珍等: "食用海藻中α-葡萄糖苷酶抑制剂的筛选及抑制动力学", 《食品工业》 * |
| 霍玉书等: "海产中药—鼠尾藻、角叉菜降糖作用的实验研究", 《中国中医药科技》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114276471A (en) * | 2021-12-30 | 2022-04-05 | 江苏海洋大学 | Preparation method and application of caulerpa lentillifera polysaccharide |
| CN115813958A (en) * | 2022-11-17 | 2023-03-21 | 江苏海洋大学 | Application of carrageenin in preparing medicine for treating Alzheimer disease and preparation method thereof |
| CN115813958B (en) * | 2022-11-17 | 2023-11-10 | 江苏海洋大学 | Application of carrageenan polyphenols in preparing medicament for treating Alzheimer's disease and preparation method thereof |
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Application publication date: 20201009 |