CN110559332A - Ganoderma lucidum and grifola frondosa hypoglycemic composition as well as preparation method and application thereof - Google Patents
Ganoderma lucidum and grifola frondosa hypoglycemic composition as well as preparation method and application thereof Download PDFInfo
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- CN110559332A CN110559332A CN201910962676.1A CN201910962676A CN110559332A CN 110559332 A CN110559332 A CN 110559332A CN 201910962676 A CN201910962676 A CN 201910962676A CN 110559332 A CN110559332 A CN 110559332A
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- China
- Prior art keywords
- extract
- grifola frondosa
- ganoderma lucidum
- solution
- bitter gourd
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- 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
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Abstract
The invention relates to a composition, in particular to a composition which has the effect of reducing blood sugar and takes ganoderma lucidum, grifola frondosa and bitter gourd as main raw materials, a preparation method and application thereof, wherein the composition is prepared from the following raw materials in percentage by weight: 20-40% of lucid ganoderma, 40-60% of grifola frondosa and 10-30% of bitter gourd. The invention has the beneficial effects that: (1) the edible fungi and the natural hypoglycemic substances are compounded, so that the effects of the raw materials in the aspect of reducing blood sugar are fully exerted, and the edible fungi has a good coordination effect of reducing blood sugar. (2) The microfiltration and ultrafiltration interception technologies adopted in the preparation method have the characteristics of simple operation, low energy consumption, short production period and the like, and can separate and protect active ingredients in the grifola frondosa extract to the maximum extent and remove ingredients with poor efficacy. (3) Animal experiment research shows that the hypoglycemic agent has obvious hypoglycemic effect.
Description
Technical Field
The invention relates to a composition, in particular to a composition which has the effect of reducing blood sugar and takes ganoderma lucidum, grifola frondosa and bitter gourd as main raw materials, a preparation method and application thereof.
Background
with the rapid development of economy, the change of life style and the acceleration of aging process, the prevalence rate of diabetes in China is in a rapid rising trend, and the diabetes becomes another chronic non-infectious disease which seriously harms the health of people after cardiovascular and cerebrovascular diseases and tumors. Diabetes is clinically mainly manifested as disorder of carbohydrate metabolism, characterized by hyperglycemia and protein glycosylation, and hyperglycemia is the main marker. The common symptoms include polydipsia, diuresis, polyphagia, emaciation and the like. The increase of blood sugar causes osmotic diuresis, polydipsia and polydipsia, and simultaneously reduces the utilization rate of the blood sugar, thereby stimulating the hunger center to cause diabetes patients to have polyphagia and hunger, and also seriously affecting the synthesis of sugar, fat and protein to cause the decomposition of the sugar, fat and protein to increase and cause emaciation. Chronic complications of diabetes, including diabetic nephropathy, diabetic microangiopathy, diabetic neuropathy, etc., which are also important causes of fatal disability of diabetes, may result from chronic non-treatment or improper treatment. Diabetes and its complications have become a worldwide public health problem seriously harming human health, and have attracted high attention from countries in the world.
Type II diabetes is the most common type of diabetes in clinic, and currently, type II diabetes reaches more than 5 hundred million patients worldwide, while china occupies about 1/3 of the patients. In addition, a large number of pre-diabetic people, namely people who have fasting plasma glucose or (and) have an increased plasma glucose after 2h of an oral glucose tolerance test but do not reach the diagnosis standard of diabetes, exist. According to the report of epidemiological investigation of adults over 18 years old in China, which is completed in 2010, the prevalence rate of diabetes in the early stage in China is 50.1 percent, namely, more than 7 hundred million of people need to perform effective intervention on blood sugar.
At present, the injection of insulin and oral drugs is the main means for treating diabetes, and the effect is also obvious, but the existing drugs, particularly western medicines, are generally accompanied by side effects, and particularly damage is caused to blood vessels, retina and nerve tissues. For the diagnosed diabetes patients, the hypoglycemic medicament should be the first choice, and the hypoglycemic medicament can be assisted by taking safe and health-care food or functional food with no side effect and definite effect. For people in early stage of diabetes mellitus, the Chinese 2-type diabetes mellitus prevention and treatment guideline recommends that patients in early stage of diabetes mellitus should reduce the risk of diabetes mellitus by diet control and exercise, and does not recommend means for preventing diabetes mellitus by using medicaments, so that the blood sugar-reducing health-care food or functional food is the first choice for the people.
Ganoderma is a large-scale fungus of Ganoderma of Ganodermataceae, and has a reputation of Mesona chinensis, and its medicinal part fruiting body has been used for more than two thousand years in China, and has effects of invigorating qi, tranquilizing mind, relieving cough and asthma, giddiness, insomnia, cardiopalmus, short breath, asthenia, cough and asthma, nourishing and strengthening body resistance, and consolidating constitution. Researches show that the indexes of blood sugar, cholesterol, triglyceride, low-density protein and the like of diabetic mice can be reduced by ganoderma lucidum, ganoderma lucidum crude polysaccharide, ganoderma lucidum aqueous extract and the like, and the ganoderma lucidum aqueous extract and the ganoderma lucidum polysaccharide have the function of reducing the blood sugar.
The fruit bodies of the grifola frondosa are soft in meat quality, are crisp like magnolia, like shredded chicken in taste, delicious in taste, unique in fragrance and rich in nutrition, are high-grade rare edible fungi, and meanwhile, the grifola frondosa is a very precious medicinal fungus and has remarkable effects of resisting tumors, reducing blood sugar, resisting hepatitis, resisting HIV viruses, improving immune system functions and the like. Researches show that both the Grifola frondosa fruiting body and Grifola frondosa polysaccharide can obviously reduce the blood sugar level of diabetic mice, and the good blood sugar reducing effect makes the Grifola frondosa fruiting body and the Grifola frondosa polysaccharide have the potential for developing blood sugar reducing medicines.
The balsam pear is sweet and bitter in fruit taste, mature pulp and aril are mainly used as vegetables for eating, and meanwhile, the balsam pear is a traditional Chinese medicine and food dual-purpose plant, is rich in a plurality of active ingredients such as balsam pear polysaccharide, saponin, polypeptide and flavonoid compounds, and has health care functions of assisting in reducing blood sugar, reducing blood fat, resisting oxidation, enhancing immunity, preventing obesity and the like. Research shows that the balsam pear extract can obviously reduce the blood sugar level of diabetic rats, improve the SOD content and the insulin level, and reduce the triglyceride and the cholesterol level. The balsam pear extract has the functions of reducing blood sugar and blood fat of hyperglycemic animals.
At present, the utilization effect of the ganoderma lucidum, the grifola frondosa and the balsam pear is not expected in the prior art, and the report of preparing the hypoglycemic health-care food or medicament by adopting the ganoderma lucidum, the grifola frondosa and the balsam pear is not available.
Disclosure of Invention
Based on the composition, the invention provides a composition with the efficacy of reducing blood sugar, and a preparation method and application thereof.
the invention achieves the above purposes through the following scheme:
In a first aspect, the invention provides a ganoderma lucidum and grifola frondosa hypoglycemic composition, which is prepared from the following raw materials in percentage by weight: 20-40% of lucid ganoderma, 40-60% of grifola frondosa and 10-30% of bitter gourd.
Preferably, the grifola frondosa in the composition is treated by the following method: taking a grifola frondosa raw material, adding 8-20 times of water by weight, heating to 80-105 ℃, extracting for 1-3 times, wherein the single extraction time is 1-4 hours, filtering the extracting solutions, combining, concentrating until the solid content is 5% -20%, obtaining a grifola frondosa extract concentrated solution, performing microfiltration treatment on the grifola frondosa extract concentrated solution, collecting microfiltration permeating solution, performing ultrafiltration on the microfiltration permeating solution, performing interception purification on the extracted components by using an ultrafiltration membrane, wherein the pore size of the ultrafiltration membrane corresponds to the molecular weight intercepted by 1-10KDa, the operating pressure is 3-8MPa, the temperature is 25-40 ℃, retaining ultrafiltration intercepting solution which does not permeate through the membrane, obtaining grifola frondosa extract ultrafiltration intercepting solution, and drying the intercepting solution to obtain the grifola frondosa extract.
Preferably, the concentration is preferably vacuum concentration.
Preferably, the Grifola frondosa raw material is Grifola frondosa fruiting body.
Preferably, the drying comprises one or more of spray drying, freeze drying and vacuum drying.
Preferably, the ganoderma lucidum in the composition is treated by the following method: adding 8-40 times of water into Ganoderma lucidum raw material, extracting at 80-105 deg.C for 1-3 times, each time for 2-6 hr, filtering the extractive solutions, mixing, concentrating to solid content of 5% -25% to obtain Ganoderma lucidum extract concentrate, and drying the Ganoderma lucidum extract concentrate to obtain Ganoderma lucidum extract.
preferably, the bitter gourds in the composition are treated by the following method: taking a bitter gourd raw material, adding 8-20 times of water by weight, extracting for 1-3 times at 80-105 ℃, wherein the extraction time is 1-4 hours each time, filtering the extracting solution, combining, concentrating to the solid content of 5% -25% to obtain a bitter gourd extract concentrated solution, and drying the bitter gourd extract concentrated solution to obtain the bitter gourd extract.
Preferably, the ganoderma lucidum and the momordica charantia in the composition are treated by the following method: mixing Ganoderma and fructus Momordicae Charantiae, adding 8-40 times of water, extracting at 80-105 deg.C for 1-3 times, each for 1-4 hr, filtering the extractive solutions, mixing, concentrating to solid content of 5% -25% to obtain Ganoderma and fructus Momordicae Charantiae extract concentrate, and drying to obtain Ganoderma and fructus Momordicae Charantiae extract.
Preferably, the above extracts can also be made into dry powder or granule.
further preferably, the prepared granules can be prepared into granules by adding auxiliary materials and performing wet granulation or one-step granulation.
Preferably, the ganoderma lucidum raw material is ganoderma lucidum coarse powder.
Preferably, the bitter gourd is a dried bitter gourd slice product.
Preferably, the concentration is a vacuum concentration.
Preferably, the drying comprises one or more of spray drying, freeze drying and vacuum drying.
according to the invention, the lucid ganoderma, the grifola frondosa and the bitter gourd in a specific ratio are compounded for use, so that compared with the single use, the effect of reducing blood sugar is obviously improved, and compared with other edible fungi or fruit and vegetable ratios, the effect of reducing blood sugar is more obvious.
In a second aspect, the invention further provides a preparation method of the ganoderma lucidum and grifola frondosa hypoglycemic composition, which comprises the following steps:
(1) Preparing a grifola frondosa extract: taking a grifola frondosa raw material, adding 8-20 times of water by weight, heating to 80-105 ℃, extracting for 1-3 times, wherein the single extraction time is 1-4 hours, filtering the extracting solutions, combining, concentrating until the solid content is 5% -20%, obtaining a grifola frondosa extract concentrated solution, performing microfiltration treatment on the grifola frondosa extract concentrated solution, collecting microfiltration permeating solution, performing ultrafiltration on the microfiltration permeating solution, performing interception purification on the extracted components by using an ultrafiltration membrane, wherein the pore size of the ultrafiltration membrane corresponds to the molecular weight intercepted by 1-10KDa, the operating pressure is 3-8MPa, the temperature is 25-40 ℃, retaining ultrafiltration intercepting solution which does not permeate through the membrane, obtaining grifola frondosa extract ultrafiltration intercepting solution, and drying the intercepting solution to obtain a grifola frondosa extract;
(2) The ganoderma lucidum and bitter gourd extract: mixing a ganoderma lucidum raw material and a bitter gourd raw material, adding 8-40 times of water by weight, extracting for 1-3 times at 80-105 ℃, wherein the extraction time is 1-4 hours each time, filtering the extracting solution, combining, concentrating until the solid content is 5% -25% to obtain a ganoderma lucidum bitter gourd extract concentrated solution, and drying the ganoderma lucidum bitter gourd extract concentrated solution to obtain a ganoderma lucidum bitter gourd extract;
(3) mixing the extract of Ganoderma and fructus Momordicae Charantiae with the extract of Grifola frondosa to obtain composition;
Wherein the weight percentages of the prepared raw materials are as follows: 20-40% of lucid ganoderma, 40-60% of grifola frondosa and 10-30% of bitter gourd.
preferably, the preparation of the ganoderma lucidum and bitter gourd extract can also be realized by respectively preparing the ganoderma lucidum extract and the bitter gourd extract, and comprises the following steps:
(1) preparing a ganoderma lucidum extract: adding 8-40 times of water into Ganoderma lucidum raw material, extracting at 80-105 deg.C for 1-3 times, each time for 2-6 hr, filtering the extractive solutions, mixing, concentrating to solid content of 5% -25% to obtain Ganoderma lucidum extract concentrate, and drying the Ganoderma lucidum extract concentrate to obtain Ganoderma lucidum extract;
(2) Preparing a bitter gourd extract: taking a bitter gourd raw material, adding 8-20 times of water by weight, extracting for 1-3 times at 80-105 ℃, wherein the extraction time is 1-4 hours each time, filtering the extracting solution, combining, concentrating to the solid content of 5% -25% to obtain a bitter gourd extract concentrated solution, and drying the bitter gourd extract concentrated solution to obtain the bitter gourd extract.
Under the condition that the ganoderma lucidum extract and the bitter gourd extract are respectively prepared, the preparation method of the ganoderma lucidum and grifola frondosa hypoglycemic composition comprises the following steps:
(1) Preparing a ganoderma lucidum extract: adding 8-40 times of water into Ganoderma lucidum raw material, extracting at 80-105 deg.C for 1-3 times, each time for 2-6 hr, filtering the extractive solutions, mixing, concentrating to solid content of 5% -25% to obtain Ganoderma lucidum extract concentrate, and drying the Ganoderma lucidum extract concentrate to obtain Ganoderma lucidum extract;
(2) Preparing a bitter gourd extract: taking a bitter gourd raw material, adding 8-20 times of water by weight, extracting for 1-3 times at 80-105 ℃, wherein the extraction time is 1-4 hours each time, filtering the extracting solution, combining, concentrating to the solid content of 5% -25% to obtain a bitter gourd extract concentrated solution, and drying the bitter gourd extract concentrated solution to obtain a bitter gourd extract;
(3) Preparing a grifola frondosa extract: taking a grifola frondosa raw material, adding 8-20 times of water by weight, heating to 80-105 ℃, extracting for 1-3 times, wherein the single extraction time is 1-4 hours, filtering the extracting solutions, combining, concentrating until the solid content is 5% -20%, obtaining a grifola frondosa extract concentrated solution, performing microfiltration treatment on the grifola frondosa extract concentrated solution, collecting microfiltration permeating solution, performing ultrafiltration on the microfiltration permeating solution, performing interception purification on the extracted components by using an ultrafiltration membrane, wherein the pore size of the ultrafiltration membrane corresponds to the molecular weight intercepted by 1-10KDa, the operating pressure is 3-8MPa, the temperature is 25-40 ℃, retaining ultrafiltration intercepting solution which does not permeate through the membrane, obtaining grifola frondosa extract ultrafiltration intercepting solution, and drying the intercepting solution to obtain a grifola frondosa extract;
(4) Mixing Ganoderma extract, fructus Momordicae Charantiae extract and Grifola frondosa extract to obtain composition for lowering blood sugar;
Wherein the weight percentages of the prepared raw materials are as follows: 20-40% of lucid ganoderma, 40-60% of grifola frondosa and 10-30% of bitter gourd
Preferably, the above extracts can also be made into dry powder or granule.
Further preferably, the prepared granules can be prepared into granules by adding auxiliary materials and performing wet granulation or one-step granulation.
Preferably, the concentration is preferably vacuum concentration.
Preferably, the drying comprises one or more of spray drying, freeze drying and vacuum drying.
preferably, the Grifola frondosa raw material is Grifola frondosa fruiting body.
Preferably, the ganoderma lucidum raw material is ganoderma lucidum coarse powder.
Preferably, the bitter gourd is a dried bitter gourd slice product.
In a third aspect, a hypoglycemic composition prepared by the preparation method is provided.
In a fourth aspect, there is provided the use of the above composition for lowering or controlling blood glucose, or treating diabetes.
Preferably, the diabetes is the treatment of type II diabetes.
Preferably, said reducing or controlling blood glucose is reducing or controlling blood glucose in a pre-diabetic population
the application of the composition is used for preparing health-care food or medicine for reducing or controlling blood sugar or treating diabetes.
Preferably, the diabetes is the treatment of type II diabetes.
Preferably, said lowering or controlling blood glucose is lowering or controlling blood glucose in a pre-diabetic population.
In a preferred embodiment, the use of the above composition for the preparation of a health food for reducing or controlling blood glucose in pre-diabetic persons.
in a preferred embodiment, the composition is used for preparing health-care food or medicine for treating type II diabetes.
on the basis of the composition, food or pharmaceutically acceptable auxiliary materials are added through the addition or preparation method of the conventional auxiliary materials of various formulations to obtain health-care food or medicaments, including but not limited to tablets, powder, granules and capsules.
compared with the prior art, the invention has the beneficial effects that:
(1) the invention utilizes edible and medicinal fungi grifola frondosa, lucid ganoderma and food balsam pear with efficacy as raw materials, has the characteristics of naturalness and safety, and simultaneously, the edible fungi and natural hypoglycemic substances are compounded, so that the efficacy of all the raw materials in the aspect of reducing blood sugar is fully exerted, and the invention has good coordination effect of reducing blood sugar.
(2) The microfiltration and ultrafiltration interception technologies adopted in the preparation method have the characteristics of simple operation, low energy consumption, short production period and the like, and can separate and protect active ingredients in the grifola frondosa extract to the maximum extent and remove ingredients with poor efficacy.
(3) Animal experiment research shows that the composition has obvious blood sugar reducing effect, can obviously improve II diabetes rat model glycolipid metabolism pathological state induced by alloxan intraperitoneal injection and high-fat feed feeding, and has obvious blood sugar reducing effect.
Detailed Description
The present invention is further illustrated by the following specific examples.
example 1
a composition with the efficacy of reducing blood sugar is prepared from the following raw materials in percentage by weight: 30% of lucid ganoderma, 60% of grifola frondosa and 10% of bitter gourd.
The preparation method of the composition comprises the following steps:
(1) Taking the ganoderma lucidum and bitter gourd raw materials according to the formula amount, adding 10 times of water by weight, heating to 105 ℃, extracting for 2 times, extracting for 2 hours, filtering the extracting solution, combining, concentrating in vacuum until the solid content is 18% to obtain ganoderma lucidum and bitter gourd extract concentrated solution, and performing spray drying to obtain dried ganoderma lucidum and bitter gourd extract powder;
(2) Taking a formula amount of grifola frondosa raw material, adding 10 times of water by weight, heating to 105 ℃, extracting for 2 times, extracting for 2 hours, filtering the extracting solutions, combining, concentrating in vacuum to reach a solid content of 15%, carrying out microfiltration treatment on the grifola frondosa extract concentrated solution, collecting microfiltration permeate with a pore diameter of 100nm, controlling the temperature to be 30 +/-5 ℃, controlling the operating pressure to be less than 1.0MPa, collecting the microfiltration permeate, discarding microfiltration non-permeate, introducing the microfiltration permeate into ultrafiltration equipment, intercepting and purifying the extract components by using an ultrafiltration membrane, wherein the pore diameter of the ultrafiltration membrane corresponds to the molecular weight of 5KDa, the operating pressure is 5 +/-1 MPa, controlling the temperature to be 25-30 ℃, stopping ultrafiltration when the volume of the permeate reaches one third, and retaining the ultrafiltration retentate of the non-permeable membrane to be used as grifola extract ultrafiltration concentrated solution. Spray drying the concentrated solution to obtain Grifola frondosa extract powder.
(3) The obtained 2 kinds of extract powders were mixed uniformly to obtain the hypoglycemic composition of example 1.
Example 2
a composition with the efficacy of reducing blood sugar is prepared from the following raw materials in percentage by weight: 30% of lucid ganoderma, 60% of grifola frondosa and 10% of bitter gourd.
The preparation method of the composition comprises the following steps:
(1) Taking the ganoderma lucidum raw material with the formula amount, adding 10 times of water by weight, heating to 100 ℃, extracting for 2 times, wherein the extraction time is 2 hours each time, filtering the extracting solution, combining, concentrating in vacuum until the solid content is 15% to obtain the ganoderma lucidum bitter gourd extract concentrated solution, and performing spray drying to obtain the dried ganoderma lucidum extract;
(2) Taking bitter gourd raw materials in a formula amount, adding 10 times of water by weight, heating to 100 ℃, extracting for 2 times, wherein each extraction time is 2 hours, filtering the extracting solutions, combining, concentrating in vacuum to the solid content of 20% to obtain bitter gourd extract concentrated solution, and performing spray drying to obtain a dried bitter gourd extract;
(3) Taking a formula amount of grifola frondosa raw material, adding 15 times of water by weight, heating to 105 ℃, extracting for 2 times, extracting for 2 hours, filtering the extracting solution, combining, concentrating in vacuum to reach a solid content of 15%, carrying out microfiltration treatment on the grifola frondosa extract concentrated solution, controlling the temperature to be 25-60 ℃, controlling the operating pressure to be less than 1MPa, collecting microfiltration permeate, discarding microfiltration non-permeate, introducing the microfiltration permeate into ultrafiltration equipment, carrying out interception and purification on the extract component by using an ultrafiltration membrane, controlling the temperature to be 25-40 ℃, stopping ultrafiltration when the permeate reaches a quarter volume, retaining the ultrafiltration retentate of the non-permeate as grifola extract ultrafiltration concentrated solution, and carrying out spray drying on the concentrated solution to obtain the grifola frondosa extract.
(3) the 3 extracts were mixed well to obtain the hypoglycemic composition of example 2.
Example 3
A composition with the efficacy of reducing blood sugar is prepared from the following raw materials in percentage by weight: 20% of lucid ganoderma, 50% of grifola frondosa and 30% of balsam pear.
The preparation method of the composition comprises the following steps:
(1) taking the ganoderma lucidum and bitter gourd raw materials according to the formula amount, adding 20 times of water by weight, heating to 100 ℃, extracting for 2 times, wherein the extraction time is 4 hours, filtering the extracting solution, combining, and concentrating in vacuum until the solid content is 8% to obtain the ganoderma lucidum and bitter gourd extract concentrated solution.
(2) taking a formula amount of grifola frondosa raw material, adding 15 times of water by weight, heating to 105 ℃, extracting for 2 times, extracting for 2 hours, filtering the extracting solution, combining, concentrating in vacuum to reach a solid content of 15%, carrying out microfiltration treatment on the grifola frondosa extract concentrated solution, controlling the temperature to be 25-60 ℃, controlling the operating pressure to be less than 1.0MPa, collecting microfiltration permeate, discarding microfiltration non-permeate, introducing the microfiltration permeate into ultrafiltration equipment, intercepting and purifying the extract components by using an ultrafiltration membrane, wherein the pore size of the ultrafiltration membrane corresponds to the molecular weight of 5KDa, the operating pressure is 3-8MPa, controlling the temperature to be 25-40 ℃, stopping ultrafiltration when the permeate reaches a quarter volume, and retaining the ultrafiltration retentate of the non-permeate as grifola extract ultrafiltration concentrated solution.
(3) spray drying the above Grifola frondosa ultrafiltered concentrated solution and Ganoderma and fructus Momordicae Charantiae concentrated solution respectively to obtain dried extract powder.
(4) The 2 extracts were mixed well to give the composition of example 3.
Further, the obtained extract is compounded with auxiliary materials such as isomalt, microcrystalline cellulose, magnesium stearate, silicon dioxide and the like, and the mixture is subjected to preparation process steps such as weighing, sieving, mixing, tabletting, coating and the like to prepare the tabletting candy which can be used as a health food with the auxiliary blood sugar reducing effect.
Test example 1
The composition of example 1 was subjected to preliminary measurements, and the polysaccharide content of the composition was: the polysaccharide content of the grifola frondosa extract is 4.1-4.3%, the polysaccharide content of the ganoderma lucidum extract is 15%, and the polysaccharide content of the balsam pear extract is 1.2-1.5%.
Effect example 1
The test was carried out using the composition prepared in example 2 as the subject.
1. The material and the method are as follows:
(1) Material
Inventive example 2 composition (abbreviated: GGM).
Animals: 105 SPF SD rats with body mass of 130-150g, 5-7 weeks old, animal qualification number: no. 44007200031231. The production license number is provided by Guangdong province medical experimental animal center: SCXK (Guangdong) 2013-0002. Animals were housed in the SPF animal house of the laboratory animal institute of Guangdong province, and the animal use license number: SYXK (Yue) 2016-0122. The animals used are ventilated well, and are changed for 12h in normal day and night, the relative humidity is (50 +/-5)%, and the room temperature is (22 +/-2) ° C, and the experiment is started after 1 week of adaptive quarantine observation.
Maintaining the feed: the formula of common feed; high-heat energy feed: 52.6 percent of maintenance materials for big and small mice, 15.0 percent of cane sugar, 15.0 percent of yolk powder, 10.0 percent of lard, 5.0 percent of casein, 1.2 percent of cholesterol, 0.6 percent of calcium bicarbonate, 0.4 percent of stone powder, 0.2 percent of sodium cholate and provided by the medical experimental animal center of Guangdong province.
Alloxan (ALX) Sigma usa; saline Guizhou Tiandi pharmacy, Inc.; blood glucose test paper roche diagnostic products (shanghai) limited; metformin hydrochloride tablet, shanghai shigui pharmaceutical ltd; insulin ELISA kit Beijing, Andy Huatai science and technology Co., Ltd; cholesterol determination kit, triglyceride determination kit and glycogen determination kit Nanjing are built into the bioengineering research institute.
(2) method of producing a composite material
GGM effect experiment on normal rat fasting blood sugar
after 20 SD rat animals are fasted for 3-5 hours, tail vein blood is taken, blood glucose value is measured by a glucometer, and the blood glucose value is randomly divided into two groups of a blank control group and a GGM high dose group according to the fasting blood glucose value, wherein each group comprises 10 animals. The blank control was left untreated, and the high-dose group of GGM was gavaged with 300mg/kg of GGM 1 time per day for 30 consecutive days. At the end of the experiment, fasting blood glucose values were again determined and fasting blood glucose values were compared between the two groups of animals.
animal grouping and glycolipid metabolism disorder Induction in rat model
The normal maintenance feed was adapted to feeding for 7 days, and 12 animals were randomly selected as a normal control group. Animals were fasted for 24 hours (free drinking water), and except for normal control animals, rats were injected with alloxan 105mg/kg i.p. 1mL/100g body weight. In addition, the molding animals are fed by replacing high-heat energy feed, and normal control animals are continuously and normally kept for feeding. After 5-7 days, the animals are fasted for 3-5 hours, blood is taken from the tail vein, a glucometer is used for measuring the blood sugar, and the model animals with the blood sugar value of 10-25 mmol/L are selected according to the blood sugar value to continue the experiment. Animals were randomly divided into 5 groups based on blood glucose value, 12 per group: model control group, metformin group, GGM low dose group, GGM medium dose group and GGM high dose group.
Animal administration
After grouping animals, each group of animals was gavaged with the corresponding test substance: injecting equal volume of purified water into the normal control group and the model control group according to the volume of 10 mL/kg; metformin hydrochloride with the dose of 200mg/kg is administrated in a metformin hydrochloride group (positive group) with the concentration of 10 mg/mL; GGM of which the low-dose group is administrated by the administration concentration of 3.75mg/mL according to the dose of 75 mg/kg; the GGM medium dose group is administrated with the GGM with the concentration of 7.5mg/mL according to the dose of 150 mg/kg; GGM with the concentration of 15mg/mL is administrated by the GGM high-dose group according to the dose of 300 mg/kg. For 30 consecutive days, 1 time per day.
2. Result detection
(1) Fasting blood sugar
the fasting blood glucose values when animals are grouped are taken as the fasting blood glucose values before administration to animals (A0), after animals are continuously administered for 30 days, fasting is carried out for 3-5 hours, blood is taken from tail veins for measuring blood glucose, and the fasting blood glucose values after administration to animals (A1). And calculating the fasting blood glucose change rate of each group of animals according to the following formula:
(2) Sugar tolerance test
after continuously administering to animals for 30 days, fasting for 3-5h, collecting blood from tail vein to measure blood sugar, and taking blood sugar value as blood sugar tolerance experimental animal 0h, after 15-20min, orally administering 2.5g/kg glucose to each group, and measuring blood sugar values of each group 0.5h and 2h after glucose administration as blood sugar value of 0.5h and 2h, respectively. And calculating the area under the blood glucose curve (AUC) for each group of animals according to the following formula:
(3) Cholesterol (TC) and Triglyceride (TG) contents
After the sugar tolerance experiment is completed, 2% sodium pentobarbital (40mg/kg, 2ml/kg) is used for carrying out intraperitoneal injection on the anesthetized animals, blood is taken from an abdominal aorta, after the blood is kept still for 0.5h, the blood is centrifuged for 15min at 3500rpm and 4 ℃, the blood serum is separated, and the TC and TG contents in the blood serum are detected according to the requirements of the kit specification.
(4) Glycogen content of liver
After the animal blood is taken, the liver is quickly removed and weighed, partial tissues of the liver large leaf are cut out, and 1: 9, centrifuging at 4 ℃ and 5000rpm for 20min, taking the supernatant, and detecting the glycogen content in the liver homogenate according to the requirements of a kit specification.
(5) Data analysis
All data are expressed as means plus minus standard deviation. The average numbers among the groups are compared by adopting One-Way ANOVA (One-Way ANOVA), the average numbers among the groups are compared pairwise, and an LSD (least squares) method is adopted when the variance is uniform; dunnett's T3 method is used when variance is irregular. Finished by SPSS l9.0 software, α ═ 0.05.
3. Results and analysis
(1) Effect of GGM on body weight and fasting plasma glucose of Normal animals
The results of the experiments on the effect of GGM on fasting plasma glucose in normal rats are shown in tables 1 and 2.
TABLE 1 Effect of GGM on body weight of Normal animals (n-10)
Note: compared with blank group, delta p is less than 0.05, and delta p is less than 0.01.
TABLE 2 Effect of GGM on fasting plasma glucose in Normal rats (n ═ 10)
note: compared with blank group, delta p is less than 0.05, and delta p is less than 0.01.
As can be seen from tables 1 and 2, there was no significant difference in the body weight change of the animals in the GGM group (p > 0.05) compared with the animals in the blank control group; there was also no significant difference in the fasting glucose values at the end of the experiment (p > 0.05).
(2) Effect of GMM on model animal weight
The results are shown in Table 3.
Table 3 animal weight change (n as 12)
Note: in contrast to the model set,*p＜0.05，**p＜0.01。
As can be seen from Table 3, the body weight of the model animals was significantly reduced before the end of the experiment compared with the normal control animals, and the difference was statistically significant (p < 0.01). Compared with model animals, the weight of animals in low, medium and high dose groups of GGM is obviously increased before the experiment is finished, and the difference statistical significance (p is less than 0.05 or p is less than 0.01) shows that the weight of rats with glycolipid metabolic disturbance has obvious reduction trend, but the weight reduction effect of model animals can be improved by GGM.
(3) Effect of GGM on fasting plasma glucose in model animals
The results are shown in Table 4.
TABLE 4 fasting plasma glucose assay results for each group of animals (n ═ 12)
note: p < 0.05, p < 0.01, compared to model groups.
As can be seen from Table 4, the fasting blood glucose value of the mice after molding is significantly higher than that of the normal control animals (p < 0.01). After the animals in each group are treated by administration for 30 days, the blood sugar value of each administration group has a descending trend of different degrees: the blood sugar reduction rate of the positive control group is (36.34 +/-6.97)%, and the blood sugar reduction rate of the low, medium and high-dose GGM groups is (18.39 +/-5.64)%, (25.57 +/-3.69)%, and (39.62 +/-3.06)%, respectively. The results show that GGM has obvious hypoglycemic effect.
(4) Effect of GGM on oral glucose tolerance in model animals
30 days after the animal is administrated, 2.5g/kg of glucose is orally taken, the blood sugar values of the animal at 0h, 0.5h and 2h are measured, the area under the blood sugar curve (AUC) is calculated, and the smaller the AUC is, the faster the recovery capability of the blood sugar level is; the greater the AUC, the slower the glycemic recovery. The results of the glucose tolerance test are shown in table 5.
Table 5 results of sugar tolerance test (n ═ 12)
Note: in contrast to the model set,*p＜0.05，**p＜0.01。
as can be seen from Table 5, compared with the normal control animals, the blood glucose value of the model control animals rapidly rises and slowly recovers after the oral administration of glucose, the AUC is obviously increased, and the difference has statistical significance (p is less than 0.01). Compared with model control animals, after fasting glucose, the blood glucose recovery of animals of each dose group of GGM is faster, the AUC is obviously reduced, and the difference has statistical significance (p is less than 0.05 or p is less than 0.01). The results show that GGM can obviously improve the glucose tolerance of rats.
(5) Effect of GGM on blood lipids of model animals
the results are shown in Table 6.
table 6 serum TC and TG measurement results of each group of animals (n-12)
Note: in contrast to the model set,*p＜0.05，**p＜0.01。
As shown in Table 6, after the animal alloxan is injected into the abdominal cavity and fed with high-fat feed for molding, the TC and TG levels in the animal serum are obviously increased compared with those in the normal control animal (p is less than 0.01). Serum TC and TG levels were reduced in each group administered GGM: compared with a model control group, the TC and TG of the GGM low-dose group have a certain descending trend, but the difference has no statistical significance (p is more than 0.05); the TC and TG of the dose group in the GGM are both obviously reduced, and the difference has statistical significance (p is less than 0.05 or p is less than 0.01); TC and TG in the GGM high-dose group are both reduced remarkably, and the difference has statistical significance (p is less than 0.01). The results show that GGM has obvious blood fat reducing effect on rats with glycolipid metabolism disorder.
(6) Effect of GGM on hepatic glycogen synthesis in model animals
The results are shown in Table 7.
Table 7 glycogen assay results of animal livers of each group (n-12)
Note: in contrast to the model set,*p＜0.05，**p＜0.01。
As can be seen from the results in Table 7, hepatic glycogen synthesis in the model control group animals was significantly decreased compared to the normal group animals, and the difference was statistically significant (p < 0.01); compared with model control animals, the glycogen content of the liver of animals in each dose administration group of GGM tends to increase, wherein the high and medium dose groups of GGM are obviously increased, and the difference has statistical significance (p is less than 0.05 or p is less than 0.01). The results show that GGM has the function of promoting hepatic glycogenesis of rats with glycolipid metabolic disorders.
4. conclusion
The composition of the grifola frondosa, the balsam pear and the ganoderma lucidum extract can obviously improve the glucose and lipid metabolism pathological state of a diabetes rat model induced by intraperitoneal injection of alloxan and feeding of high-fat feed, and has good application prospect in the development of functional products for reducing blood sugar and blood lipid.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and their concepts should be considered to be equivalent or modified within the technical scope of the present invention.
Claims (10)
1. The composition for reducing blood sugar of ganoderma lucidum and grifola frondosa is characterized by being prepared from the following raw materials in percentage by weight: 20-40% of lucid ganoderma, 40-60% of grifola frondosa and 10-30% of bitter gourd.
2. The composition as claimed in claim 1, wherein the grifola frondosa in the composition is treated by the following method: taking a grifola frondosa raw material, adding 8-20 times of water by weight, heating to 80-105 ℃, extracting for 1-3 times, wherein the single extraction time is 1-4 hours, filtering the extracting solutions, combining, concentrating until the solid content is 5% -20%, obtaining a grifola frondosa extract concentrated solution, performing microfiltration treatment on the grifola frondosa extract concentrated solution, collecting microfiltration permeating solution, performing ultrafiltration on the microfiltration permeating solution, performing interception purification on the extracted components by using an ultrafiltration membrane, wherein the pore size of the ultrafiltration membrane corresponds to the molecular weight intercepted by 1-10KDa, the operating pressure is 3-8MPa, the temperature is 25-40 ℃, retaining ultrafiltration intercepting solution which does not permeate through the membrane, obtaining grifola frondosa extract ultrafiltration intercepting solution, and drying the intercepting solution to obtain the grifola frondosa extract.
3. The composition according to claim 1, wherein the ganoderma lucidum in the composition is treated by the following method: adding 8-40 times of water into Ganoderma lucidum raw material, extracting at 80-105 deg.C for 1-3 times, each time for 2-6 hr, filtering the extractive solutions, mixing, concentrating to solid content of 5% -25% to obtain Ganoderma lucidum extract concentrate, and drying the Ganoderma lucidum extract concentrate to obtain Ganoderma lucidum extract.
4. A composition according to claim 1 wherein the goya in the composition is treated by a method comprising: taking a bitter gourd raw material, adding 8-20 times of water by weight, extracting for 1-3 times at 80-105 ℃, wherein the extraction time is 1-4 hours each time, filtering the extracting solution, combining, concentrating to the solid content of 5% -25% to obtain a bitter gourd extract concentrated solution, and drying the bitter gourd extract concentrated solution to obtain the bitter gourd extract.
5. the composition according to claim 1, wherein the ganoderma lucidum and momordica charantia in the composition are treated by the method comprising: mixing Ganoderma and fructus Momordicae Charantiae, adding 8-40 times of water, extracting at 80-105 deg.C for 1-3 times, each for 1-4 hr, filtering the extractive solutions, mixing, concentrating to solid content of 5% -25% to obtain Ganoderma and fructus Momordicae Charantiae extract concentrate, and drying to obtain Ganoderma and fructus Momordicae Charantiae extract.
6. A preparation method of a ganoderma lucidum and grifola frondosa hypoglycemic composition is characterized by comprising the following steps:
(1) Preparing a grifola frondosa extract: taking a grifola frondosa raw material, adding 8-20 times of water by weight, heating to 80-105 ℃, extracting for 1-3 times, wherein the single extraction time is 1-4 hours, filtering the extracting solutions, combining, concentrating until the solid content is 5% -20%, obtaining a grifola frondosa extract concentrated solution, performing microfiltration treatment on the grifola frondosa extract concentrated solution, collecting microfiltration permeating solution, performing ultrafiltration on the microfiltration permeating solution, performing interception purification on the extracted components by using an ultrafiltration membrane, wherein the pore size of the ultrafiltration membrane corresponds to the molecular weight intercepted by 1-10KDa, the operating pressure is 3-8MPa, the temperature is 25-40 ℃, retaining ultrafiltration intercepting solution which does not permeate through the membrane, obtaining grifola frondosa extract ultrafiltration intercepting solution, and drying the intercepting solution to obtain a grifola frondosa extract;
(2) The ganoderma lucidum and bitter gourd extract: mixing a ganoderma lucidum raw material and a bitter gourd raw material, adding 8-40 times of water by weight, extracting for 1-3 times at 80-105 ℃, wherein the extraction time is 1-4 hours each time, filtering the extracting solution, combining, concentrating until the solid content is 5% -25% to obtain a ganoderma lucidum bitter gourd extract concentrated solution, and drying the ganoderma lucidum bitter gourd extract concentrated solution to obtain a ganoderma lucidum bitter gourd extract;
(3) Mixing the extract of Ganoderma and fructus Momordicae Charantiae with the extract of Grifola frondosa to obtain composition;
Wherein the weight percentages of the prepared raw materials are as follows: 20-40% of lucid ganoderma, 40-60% of grifola frondosa and 10-30% of bitter gourd.
7. A preparation method of a ganoderma lucidum and grifola frondosa hypoglycemic composition is characterized by comprising the following steps:
(1) Preparing a ganoderma lucidum extract: adding 8-40 times of water into Ganoderma lucidum raw material, extracting at 80-105 deg.C for 1-3 times, each time for 2-6 hr, filtering the extractive solutions, mixing, concentrating to solid content of 5% -25% to obtain Ganoderma lucidum extract concentrate, and drying the Ganoderma lucidum extract concentrate to obtain Ganoderma lucidum extract;
(2) Preparing a bitter gourd extract: taking a bitter gourd raw material, adding 8-20 times of water by weight, extracting for 1-3 times at 80-105 ℃, wherein the extraction time is 1-4 hours each time, filtering the extracting solution, combining, concentrating to the solid content of 5% -25% to obtain a bitter gourd extract concentrated solution, and drying the bitter gourd extract concentrated solution to obtain a bitter gourd extract;
(3) Preparing a grifola frondosa extract: taking a grifola frondosa raw material, adding 8-20 times of water by weight, heating to 80-105 ℃, extracting for 1-3 times, wherein the single extraction time is 1-4 hours, filtering the extracting solutions, combining, concentrating until the solid content is 5% -20%, obtaining a grifola frondosa extract concentrated solution, performing microfiltration treatment on the grifola frondosa extract concentrated solution, collecting microfiltration permeating solution, performing ultrafiltration on the microfiltration permeating solution, performing interception purification on the extracted components by using an ultrafiltration membrane, wherein the pore size of the ultrafiltration membrane corresponds to the molecular weight intercepted by 1-10KDa, the operating pressure is 3-8MPa, the temperature is 25-40 ℃, retaining ultrafiltration intercepting solution which does not permeate through the membrane, obtaining grifola frondosa extract ultrafiltration intercepting solution, and drying the intercepting solution to obtain a grifola frondosa extract;
(4) Mixing Ganoderma extract, fructus Momordicae Charantiae extract and Grifola frondosa extract to obtain composition for lowering blood sugar;
wherein the weight percentages of the prepared raw materials are as follows: 20-40% of lucid ganoderma, 40-60% of grifola frondosa and 10-30% of bitter gourd.
8. The method of claim 6 or 7, wherein each of the extracts is prepared as a dry powder or as a granule; preferably, the prepared granules are prepared into granules by adding auxiliary materials and performing wet granulation or one-step granulation.
9. The method of claim 6 or 7, wherein the drying comprises one or more of spray drying, freeze drying, and vacuum drying.
10. Use of the composition of any one of claims 1 to 5 for the preparation of a health food or a pharmaceutical product for lowering or controlling blood glucose or treating diabetes; preferably, the diabetes is the treatment of type II diabetes; preferably, said lowering or controlling blood glucose is lowering or controlling blood glucose in a pre-diabetic population.
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