CN115919881A - Composition with effects of reducing blood sugar and blood fat and application thereof - Google Patents
Composition with effects of reducing blood sugar and blood fat and application thereof Download PDFInfo
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- CN115919881A CN115919881A CN202211474665.7A CN202211474665A CN115919881A CN 115919881 A CN115919881 A CN 115919881A CN 202211474665 A CN202211474665 A CN 202211474665A CN 115919881 A CN115919881 A CN 115919881A
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Abstract
The invention discloses a composition with the effects of reducing blood sugar and blood fat and application thereof, wherein the active ingredients of the composition comprise berberine and digitoxin D, and the mass ratio of the berberine to the digitoxin D is 30-50. The composition comprises two monomer components of berberine and digitoxin D, and is prepared by combining according to a specific dosage. Experiments show that the composition has better effect on reducing blood sugar and blood fat in a cell layer compared with the use effect of a single compound. The two monomers generate a certain synergistic effect, can be used as related active ingredients to synergistically slow down the course of disease caused by insulin resistance of type 2 diabetes, and has a certain medicinal prospect. In addition, the two monomer compounds shown in the combination have the advantages of safety, low toxicity, wide sources and the like, and are suitable for standardized popularization and use.
Description
Technical Field
The invention relates to the technical field of medicines, and particularly relates to a composition with effects of reducing blood sugar and blood fat and application thereof.
Background
According to the 8 th edition of the global diabetes map report published by the international diabetes union (IDF), about 4.25 hundred million adults suffer from diabetes in 2017 globally, and the worldwide diabetic patients may reach 6.29 hundred million by 2045 years.
Diabetes is a series of metabolic disorder syndromes of sugar, protein, fat, water, electrolyte and the like caused by hypofunction of pancreatic islets of the body, insulin resistance and the like caused by various pathogenic factors such as genetic factors, immune dysfunction, microbial infection, mental factors and the like, and is clinically characterized by hyperglycemia, and typical cases can show polyuria, polydipsia, polyphagia and the like. In addition, diabetes may also cause complications such as infection, heart disease, cerebrovascular disease, renal failure, and binocular blindness. The drug therapy for diabetes mainly takes oral hypoglycemic drugs matched with insulin as main drugs at present, and the commonly used oral hypoglycemic drugs comprise biguanides, sulfonylureas, alpha-glucosidase inhibitors, thiazolidinediones, carbamoylmethyl benzenesulfonic acid derivatives and the like, and the diabetes is treated by correcting glucose metabolism disorder, promoting islet function recovery, improving insulin resistance and the like. However, the drugs need to be added gradually when used for a long time, and most of the drugs have adverse reactions of different degrees.
In the prior art, a single compound has a plurality of traditional Chinese medicine active ingredients with the function of reducing blood sugar, but the effect of reducing blood sugar can be achieved well by using the single compound, and how to select the compound according to the types of diseases and reasonable proportion to improve the content of active ingredients and remove ineffective ingredients is very important.
In view of this, the invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a composition with the effects of reducing blood sugar and blood fat and application thereof, the composition can remarkably slow down the course of disease caused by insulin resistance of type 2 diabetes, and has the advantages of safety, low toxicity, wide source and the like.
The invention is realized by the following steps:
the invention provides a composition with the effects of reducing blood sugar and blood fat, and the active ingredients of the composition comprise berberine and digitoxin D;
the mass ratio of the berberine to the digitoxin D is 30-50.
In an alternative embodiment, the mass ratio of berberine to digitoxin D is 35-45.
In an alternative embodiment, berberine and digitoxin D are both alcohol extracts.
In an alternative embodiment, the preparation method of the composition comprises mixing berberine and digitoxin D according to a mass ratio.
The invention provides a medicinal preparation with the effects of reducing blood sugar and blood fat, which comprises the composition.
In alternative embodiments, the dosage form of the pharmaceutical formulation is a solid formulation or a liquid formulation.
In alternative embodiments, the solid formulation is a tablet, capsule or granule; the liquid preparation is syrup, suspension or injection.
The invention also provides application of the composition in preparing a medicine or health-care product for relieving insulin resistance, which comprises the regulation and control of the expression of PIK3R1 and AKT1 genes in cells.
The invention also provides application of the composition in preparing a medicine or health-care product for treating diabetes, which comprises regulating and controlling the expression of PIK3R1 and AKT1 genes in cells.
In an alternative embodiment, the diabetes is type 2 diabetes.
The invention has the following beneficial effects:
the composition comprises two monomer components of berberine and digitoxin D, and is prepared by combining according to a specific dosage. Experiments show that the composition has better effect on reducing blood sugar and blood fat in a cell layer compared with the use effect of a single compound. The two monomers generate a certain synergistic effect, can be used as related active ingredients to synergistically slow down the course of disease caused by insulin resistance of type 2 diabetes, and has a certain medicinal prospect. In addition, the two monomer compounds shown in the combination have the advantages of safety, low toxicity, wide sources and the like, and are suitable for standardized popularization and use.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 shows the cell activity of the composition of example 1 at various concentrations;
FIG. 2 is a graph showing glucose consumption of glucosamine-induced IR-HepG2 cells at different concentrations of drugs in the experimental examples;
FIG. 3 shows the expression of AKT1 at the gene level in R-HepG2 cells in the experimental examples;
FIG. 4 shows FOXO1 expression at the gene level of R-HepG2 cells in the experimental examples;
FIG. 5 shows NF- κ B expression at the gene level of R-HepG2 cells in experimental examples.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The traditional Chinese medicine considers that the pathogenesis of the diabetes is yin deficiency and dryness heat, the yin deficiency is taken as the basis, the dryness heat is taken as the secondary aspect, the upper elimination treatment on the lung, the middle elimination treatment on the stomach and the lower elimination treatment on the kidney are advocated, and the treatment is performed from the lung, the stomach and the kidney triple energizer; furthermore, sour-bitter can soften liver and promote the production of body fluid without lingering pathogen, so as to soften bitter taste to extinguish endogenous wind, and nourish yin slowly and sweet. The invention mixes berberine and digitoxin D according to a certain proportion to prepare a composition.
The invention provides a composition with the effects of reducing blood sugar and blood fat, and the active ingredients of the composition comprise berberine and digitoxin D; the mass ratio of the berberine to the digitoxin D is 30-50.
Berberine, also known as berberine, is a quaternary ammonium alkaloid separated from Chinese medicinal rhizoma Coptidis, and is the main antibacterial active ingredient of rhizoma Coptidis.
Berberine has antibacterial effect on hemolytic streptococcus, staphylococcus aureus, gonococcus, shigella flexneri and Shigella shigella, has effect in enhancing phagocytosis of leukocyte, has inhibitory effect on tubercle bacillus and plague bacillus to different extent, and has inhibitory effect on rat amoeba. Berberine has anti-curare toxic effect on animals, and has peripheral blood pressure lowering and antipyretic effects. Berberine hydrochloride (commonly called berberine hydrochloride) has been widely used for treating gastroenteritis, bacillary dysentery, etc., and also has certain curative effect on pulmonary tuberculosis, scarlet fever, acute tonsillitis and respiratory tract infection. In traditional Chinese medicine, coptis, phellodendron, barberry root, chinese mahonia and the like are commonly used as heat-clearing and detoxifying drugs, wherein the main effective component is berberine.
The rehmannia root glycoside D is an iridoid glycoside chemical component from fresh rehmannia root, radix rehmanniae, prepared rehmannia root, rehmannia root callus, rehmannia leaf and diseased rehmannia root, and is usually used as a quality control standard substance of rehmannia root in rehmannia root medicinal materials and preparations because the pharmacodynamic action of the rehmannia root glycoside D is consistent with that of traditional Chinese medicine, the content is higher and more stable. Meanwhile, as a traditional Chinese medicine extract, the digitonin D can obviously increase the white blood cell number, the platelet number, the reticulocyte number, the bone marrow DNA content and the body weight of a blood deficiency model mouse. In addition, the rehmannia root glycoside D also has the functions of nourishing yin, regulating immunity, enriching blood and the like.
The adding proportion of the berberine and the digitoxin D is further optimized by the inventor, and in a preferred embodiment, the mass ratio of the berberine to the digitoxin D is 35-45. More preferably, the mass ratio of berberine to digitoxin D is 40.
In some embodiments, berberine and digitonin D are both alcohol extracts, in particular berberine is an ethanol extract and digitonin D is a methanol extract. However, berberine and digitoxin D can also be obtained by other extraction methods, and are not limited to alcohol extraction. In addition, the sources of the above raw materials are not particularly limited in the present invention, and conventional commercially available products known to those skilled in the art may be used.
In some embodiments, the above composition is prepared by mixing berberine and digitoxin D at a mass ratio.
The invention provides a medicinal preparation with the effects of reducing blood sugar and blood fat, which comprises the composition and a pharmaceutically acceptable carrier.
The pharmaceutically acceptable carrier in the present invention refers to pharmaceutically acceptable carriers well known to those skilled in the art, and the pharmaceutically acceptable carriers of the present invention include, but are not limited to: fillers, wetting agents, binders, disintegrants, lubricants, binders, glidants, taste masking agents, surfactants, preservatives, and the like.
In the above pharmaceutical compositions, the proportion of active ingredient may vary and may represent any proportion other than zero by weight of a given unit dosage form.
In some embodiments, the dosage form of the pharmaceutical formulation may be a solid formulation or a liquid formulation.
Specifically, the solid preparation can be tablets, capsules or granules; the liquid preparation is syrup, suspension or injection.
The invention also provides application of the composition in preparing a medicament or a health-care product for relieving insulin resistance, which comprises the step of regulating and controlling the expression of PIK3R1 and AKT1 genes in cells.
Insulin resistance means that the efficiency of insulin in promoting glucose uptake and utilization is reduced by various reasons, and the body complementarily secretes too much insulin to produce hyperinsulinemia to maintain the stability of blood sugar.
The invention also provides application of the composition in preparing a medicine or health-care product for treating diabetes, which comprises regulating and controlling the expression of PIK3R1 and AKT1 genes in cells. In the present invention, the diabetes is type 2 diabetes.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
The invention provides a composition with the effects of reducing blood sugar and blood fat, and the active ingredients of the composition comprise berberine and digitoxin D; the mass ratio of the berberine to the digitoxin D is 40.
Example 2
The invention provides a composition with the effects of reducing blood sugar and blood fat, and the active ingredients of the composition comprise berberine and digitoxin D; the mass ratio of the berberine to the digitoxin D is 35.
Example 3
The invention provides a composition with the effects of reducing blood sugar and blood fat, and the active ingredients of the composition comprise berberine and digitoxin D; the mass ratio of the berberine to the digitoxin D is 38.
Example 4
The invention provides a composition with the effects of reducing blood sugar and blood fat, and the active ingredients of the composition comprise berberine and digitoxin D; the mass ratio of the berberine to the digitoxin D is 42.
Example 5
The invention provides a composition with the effects of reducing blood sugar and blood fat, and the active ingredients of the composition comprise berberine and digitoxin D; the mass ratio of the berberine to the digitoxin D is 45.
Examples of the experiments
Example 1 cellular assays for lowering blood glucose and blood lipid levels of the compositions prepared
The experimental method is as follows:
(1) HepG2 cell Activity assay
HepG2 cells are planted in a 96-well plate at a density of 8000 cells/well for about 24 hours, after the cells adhere to the wall, the culture solution is discarded, and the liquor of berberine, namely the digitoxin D which is equal to 40 is added to ensure that the final concentration is 0, 3.125, 6.25, 12.5, 25, 50, 100, 200 and 400 mu M, 6 parallel wells are arranged at each concentration, and a culture medium blank group and a control group are additionally arranged. After further culturing for 24 hours, the activity of the cells was measured by the CCK-8 method.
(2) Glucose consumption test
8000 HepG2 cells per well are planted in a 96-well plate, after 24 hours of incubation in a high-sugar complete culture medium incubator, the adherent HepG2 cells are exposed to a low-sugar complete culture medium containing 18mM glucosamine for 18 hours, and then the cells are divided into a blank control group (con), a model group (mod) and a berberine high, medium and low dose group (40, 20 and 10 mu M); high, medium and low dose groups of digitoxin D (40, 20, 10 μ M); and berberine digitoxin D equal to 40 in the high, medium and low dose group (40, 20, 10 μ M); rosiglitazone group (Rglt) 20. Mu.M, treatment was performed for 24 hours. And then discarding the supernatant, adding a low-sugar basal medium containing 10nM insulin, placing the mixture into an incubator for incubation for 6 hours, simultaneously measuring the activity of cells in each hole, taking 1.5 mu l of the supernatant, and detecting the glucose content in the supernatant by using a glucose kit, thereby calculating the glucose consumption of each group.
Glucose content of each group of supernatants = (sample well-blank well)/(standard well-blank well) × standard concentration (5.55 mM/L); glucose consumption per group = (5.55 mM/L-supernatant content); glucose consumption (corrected) = (5.55 mM/L-supernatant content)/cell viability.
(3) Real-time fluorescent quantitative PCR
HepG2 cells at 5X 10 5 One/well was seeded into 4 dishes per group in 6-well plates, and after 24 hours of incubation in a high-sugar complete medium incubator, adherent HepG2 cells were exposed to a low-sugar complete medium containing 18mM glucosamine for 18 hours, after which the cells were divided into a blank control group (con), a model group (mod), high-medium-low dose groups (H, M, L) each having a berberine: digitonin D equal to 40, and a rosiglitazone group (R), and treated for 24 hours. After two washes with PBS, cells were harvested, total RNA was extracted according to Trizol reagent instructions, dissolved In non-enzymatic water, reverse transcribed according to the instructions of the 5 × All-In-One RT-MasterMix kit, followed by amplification In a qPCR system using SYBR Green Mix reagent (TSINGKE) with actin as an internal reference for relative expression abundance.
(4) Statistical analysis
Data were analyzed using GraphPadPrism software 8.0 and expressed as mean ± SD. Statistical significance was determined using one-way analysis of variance (ANOVA), followed by multiple comparisons using the T μ key test, and P < 0.05 was considered statistically significant.
The results are as follows:
1. effect of the compositions on HepG2 cell Activity
The CCK-8 method measures the cell viability of the extraction site after 24 hours of administration, and as shown in FIG. 1, when the administration dose is 50. Mu.M or less, no significant cytotoxicity is observed, so that the doses of 40. Mu.M, 20. Mu.M, and 10. Mu.M were selected for the subsequent glucose consumption experiments.
2. Effect of the composition on glucose consumption by glucosamine-induced IR-HepG2 cells
As shown in FIG. 2, the IR-HepG2 cell model induced by glucosamine can be alleviated to some extent by the high, medium and low doses of 40. Mu.M, 20. Mu.M and 10. Mu.M drug solutions; compared with the single compound, the compound has better using effect, a certain synergistic effect is generated between the two monomers, and the insulin resistance of cells is improved by increasing the glucose consumption of the cells.
As shown in FIG. 3, FIG. 4 and FIG. 5, the mRNA expression of glycolipid metabolism related factors of IR-HepG2 cells is regulated, and after administration, the expression levels of PI3KR1 and AKT1 in the IR-HepG2 cells are changed, respectively, including the inhibition of the expression of FOXO1 gluconeogenesis transcription gene and NF-kappa B inflammation transcription gene, and the reduction of gluconeogenesis in the IR-HepG2 cell model induced by glucosamine and inflammatory factors generated by glucotoxicity in the cells.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A composition with the effects of reducing blood sugar and blood fat is characterized in that the active ingredients of the composition comprise berberine and digitoxin D;
the mass ratio of the berberine to the digitoxin D is 30-50.
2. The composition according to claim 1, wherein the mass ratio of berberine to digitoxin D is 35-45.
3. The composition according to claim 2, wherein berberine and digitoxin D are both alcohol extracts.
4. The composition according to claim 3, wherein the composition is prepared by mixing berberine and digitoxin D in a mass ratio.
5. A pharmaceutical preparation with effects of reducing blood sugar and blood lipid, wherein the composition of any one of claims 1-4 is contained in the pharmaceutical preparation.
6. The pharmaceutical formulation of claim 5, wherein the pharmaceutical formulation is in a solid or liquid form.
7. The pharmaceutical formulation of claim 6, wherein the solid formulation is a tablet, capsule or granule; the liquid preparation is syrup, suspension or injection.
8. Use of a composition according to any one of claims 1 to 4 in the manufacture of a medicament or nutraceutical for reducing insulin resistance, comprising modulating the expression of the PIK3R1 and AKT1 genes in a cell.
9. Use of a composition according to any one of claims 1 to 4 in the manufacture of a medicament or nutraceutical for the treatment of diabetes, comprising modulating the expression of the PIK3R1 and AKT1 genes in a cell.
10. The use according to claim 9, wherein the diabetes is type 2 diabetes.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202211474665.7A CN115919881A (en) | 2022-11-23 | 2022-11-23 | Composition with effects of reducing blood sugar and blood fat and application thereof |
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| CN202211474665.7A CN115919881A (en) | 2022-11-23 | 2022-11-23 | Composition with effects of reducing blood sugar and blood fat and application thereof |
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