CN117547522A - Application of intelligent monomer delta-juniper in preparation of diabetes medicine - Google Patents
Application of intelligent monomer delta-juniper in preparation of diabetes medicine Download PDFInfo
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- CN117547522A CN117547522A CN202311816869.9A CN202311816869A CN117547522A CN 117547522 A CN117547522 A CN 117547522A CN 202311816869 A CN202311816869 A CN 202311816869A CN 117547522 A CN117547522 A CN 117547522A
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/01—Hydrocarbons
- A61K31/015—Hydrocarbons carbocyclic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/906—Zingiberaceae (Ginger family)
- A61K36/9062—Alpinia, e.g. red ginger or galangal
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Diabetes (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Botany (AREA)
- Endocrinology (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
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- Bioinformatics & Cheminformatics (AREA)
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- Obesity (AREA)
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the technical field of south medicine, and discloses application of a nootropic monomer delta-juniper in preparation of a diabetes medicine, in particular application of nootropic monomer delta-juniper in preparation of a medicine for treating and/or preventing diabetes related diseases. The invention explores the effect of delta-juniper on improving IR and sugar metabolism by constructing an IR-HepG2 cell model. The result shows that after the IR-HepG2 cells are incubated with delta-juniper in different concentrations, the intake and consumption of glucose are obviously improved, the balance of glucose inside and outside the cells is maintained, and the effect that one of four southernwood is a nootropic monomer delta-juniper can play a role in reducing blood sugar by improving the effect of IR is proved; it was further creatively found that the effects of the administration concentrations of delta-juniper in 2.5, 10, 17.5 μm, respectively, were optimal.
Description
Technical Field
The invention belongs to the technical field of south medicines, relates to a south medicine nootropic monomer delta-juniper alkene, which can exert the effect of reducing blood sugar by improving IR effect, and in particular relates to an application of nootropic monomer delta-juniper alkene in preparing diabetes medicines.
Background
Fructus Alpinae Oxyphyllae (Latin school name: alpinia oxyphylla Miq.) is named as fructus Alpinae Oxyphyllae, fructus Alpiniae Oxyphyllae, and rhizoma Alpiniae Officinarum of Zingiberaceae. Is mainly distributed in the south China, is served as a genuine medicinal material of Hainan and is praised as one of four southern medicines in China. Intelligence-promoting and medicine-taking is carried in "De-Ji-Ben-Cao", has the functions of warming spleen, relieving diarrhea, taking saliva, warming kidney, reducing urination and stopping nocturnal emission, and is used for treating symptoms such as enuresis due to kidney deficiency, frequent urination, seminal emission, turbid urine, diarrhea due to cold in spleen, cold pain in abdomen, excessive saliva and the like. The intelligence-promoting plant is a medicinal and edible plant and has the effects of warming spleen, relieving diarrhea, controlling salivation, warming kidney, reducing urination and controlling nocturnal emission. Modern pharmacological research shows that fructus alpiniae oxyphyllae has various biological activities of regulating urination function, improving cognitive ability, resisting bacteria and tumors, improving diabetes and the like.
Diabetes Mellitus (DM) is a metabolic disease characterized by hyperglycemia, and is a complex endocrine disease caused by interaction of genetic and environmental factors, namely, a series of metabolic disorder diseases such as sugar, protein, fat, secondary water, electrolyte and the like caused by relative and/or absolute insulin secretion deficiency. Diabetes is the greatest chronic disease threatening human survival at present, the diabetes is treated by insulin in the medical field all the time, and insulin is one of human hormone, and can promote sugar in blood to enter into tissue cells such as liver and fat and realize anabolism of human body, so that the normal functions of heart, brain and muscle are maintained. Once the pancreatic cells of human endocrine insulin are damaged or the sensitivity of human body to insulin is reduced, glucose cannot be absorbed by the cells and can accumulate in blood, resulting in diabetes and various complications. Diabetics may develop impaired glucose tolerance, hyperglycemia, urinary glucose, and abnormal insulin release tests. Diabetes causes blood sugar rise, can accelerate aerobic oxidation of glucose, enhance non-enzymatic glycosylation of protein and cause lipid metabolism abnormality, and cause oxidative stress and lipid metabolism disorder to a certain extent in vivo.
At present, many researches on the intelligent chemical components and pharmacology of traditional Chinese medicines are reported, but no report exists on whether the intelligent monomer delta-juniper applied to the diabetes medicine has the efficacy of reducing blood sugar and the mechanism thereof.
Disclosure of Invention
The invention aims to provide an application of a nootropic monomer delta-juniper in preparing a diabetes medicine, research on the action of the nootropic monomer delta-juniper in the four-big south medicine in improving Insulin Resistance (IR), and provide scientific basis for the clinical application of delta-juniper in the adjuvant therapy of diabetes.
In order to achieve the above purpose, the technical scheme of the invention is as follows: provides an application of a nootropic monomer delta-juniper in preparing a medicament for treating diabetes mellitus, and the nootropic monomer delta-juniper is applied to preparing a medicament for treating and/or preventing diabetes mellitus-related diseases.
Further, the dose concentration of the intelligent monomer delta-juniper alkene is 1-200 mu M.
Further, the dosage concentration of the intelligent monomer delta-juniper alkene is 1-100 mu M.
Further, the dosage concentration of the intelligent monomer delta-juniper alkene is 1-50 mu M.
Further, the concentration of the nootropic monomer delta-juniper alkene administered is 2.5 mu M.
Further, the concentration of the intelligent monomer delta-juniper is 10 mu M.
Further, the concentration of the nootropic monomer delta-juniper is 17.5 μm.
The invention has the following beneficial effects:
the invention explores the effect of delta-juniper on improving IR and sugar metabolism by constructing an IR-HepG2 cell model. The result shows that after the IR-HepG2 cells are incubated with delta-juniper in different concentrations, the intake and consumption of glucose are obviously improved, the balance of glucose inside and outside the cells is maintained, and the effect that one of four southernwood is a nootropic monomer delta-juniper can play a role in reducing blood sugar by improving the effect of IR is proved; it was further creatively found that the effects of the administration concentrations of delta-juniper in 2.5, 10, 17.5 μm, respectively, were optimal.
Drawings
FIG. 1 is a graph comparing the effect of delta-juniper on HepG2 cell viability;
FIG. 2 is a graph comparing the effect of various concentrations of delta-juniper on glucose uptake in IR-HepG2 cells (note: CON as blank; MOD as model; ROSI as rosiglitazone (positive drug group); 2.5, 10, 17.5 as delta-juniper dosing concentration; # indicates p <0.05 compared to blank; x indicates p <0.05, p <0.01 compared to model);
FIG. 3 is a graph comparing the effect of different concentrations of delta-juniper on the glucose consumption of IR-HepG2 cells (note: CON as blank; MOD as model; ROSI as rosiglitazone (positive drug group); 2.5, 10, 17.5 as delta-juniper dosing concentration; # indicates p <0.01 compared to blank; x indicates p <0.05, p <0.01 compared to model).
Detailed Description
A further understanding of the nature and advantages of the present invention may be realized by the following detailed description. The examples provided are merely illustrative of the methods of the present invention and are not intended to limit the remainder of the disclosure in any way whatsoever. The experimental methods in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1
The delta-juniper monomer is applied to the preparation of a medicament for treating and/or preventing diabetes related diseases, wherein the delta-juniper monomer is administered at a concentration of 2.5 mu M.
Example 2
The delta-juniper alkene of the invention is applied to the preparation of a medicament for treating and/or preventing diabetes related diseases, wherein the delta-juniper alkene of the invention is administered at a concentration of 10 mu M.
Example 3
The delta-juniper monomer is applied to the preparation of a medicament for treating and/or preventing diabetes related diseases, wherein the delta-juniper monomer is administered at a concentration of 17.5 mu M.
The experimental process comprises the following steps:
1. experimental materials
1. Experimental instrument
Spectra Max19 full wavelength microplate reader (U.S.); novo-Cyte3130 flow cytometer (USA); CI-191C/. Times.carbon dioxide incubator (USA); thermo Micro 21R refrigerated high speed centrifuge (USA), L420cen medical centrifuge (Hunan instruments laboratory instruments Co., ltd.).
2. Materials and reagents
Delta-juniper (Shanghai derived leaf Biotechnology Co., # 483-76-1); hepG2 cells (Qiao Xin boat biotechnology limited in Shanghai); fetal bovine serum (ExCell Bio, FSP 500); cell Counting Kit-8 (CCK 8, APExBIO, K1018); penicillin-streptomycin (neosaimei, C125C 5); 2-NBDG (fluorescent glucose analog, APExBIO, B6035); glucose assay kit (south Beijing built bioengineering institute); rosiglitazone (Shanghai source leaf biotechnology limited, Y05F9C 54480); DMEM medium (Gibco, C11885500 BT); PBS buffer (biosharp, BL 302A); trypsin-EDTA digests (Solarbio, T1300); trypsin-free of EDTA, phenol red (Biosharp, BL 526A).
2. Experimental method
1. Cell culture
HepG2 cells were cultured in DMEM complete medium (containing 10% fetal calf serum and 1% penicillin-streptomycin) at 37℃with 5% CO 2 Is cultured in a cell culture vessel. After the number of the cells reaches about 90%, discarding the culture solution, adding 1mL of PBS solution for washing for 2 times, adding 0.25% of trypsin digestion solution for digestion, stopping digestion with a complete culture medium after 1-2 min, carrying out cell passage, and taking the cells in the logarithmic phase for subsequent experiments.
2. CCK-8 detection of cell viability
Blowing and mixing HepG2 cells to obtain cell suspension, adding into 96-well plate, and adding 1×10 cells per well 5 The individual cells were placed in a cell incubator for culturing for 24 hours. When cells were grown to 70% or more, each well was replaced with 100 μl of delta-juniper medium (1, 2.5, 5, 10, 25, 50, 100, 200 μΜ) at different concentrations and incubated for 24h, while control (zero well) and blank groups were established, each with 6 multiplex wells. After the incubation, 10. Mu.LCCK-8 reagent was added to each well and incubated in an incubator protected from light for 1h. OD values were measured at 450nm wavelength using a microplate reader and cell viability was calculated for each group.
Cell viability (%) = [ (experimental OD value-blank OD value)/(control OD value-blank OD value) ]x100%.
3. Glucose uptake assay
HepG2 cells were divided into 6 groups, each consisting of a blank (CON), model (MOD), positive control (rosiglitazone, ROSI; 25. Mu. Mol/L) and delta. -juniperazine (2.5, 10, 17.5. Mu.M), each with 3 replicates. Except that a blank group adopts a 5.5mM low sugar culture medium, hepG2 cells are incubated by adopting a 30mM high sugar culture medium, an IR cell model is established, corresponding drugs are given according to cell groups for incubation for 24 hours, then the drug-containing culture medium is discarded, PBS is used for washing for 2 times, 25 mu mol/L of 2-NBDG solution is added, the mixture is incubated at 37 ℃ in a dark place for 30 minutes, and the flow type fluorescence intensity in the HepG2 cells is detected by utilizing a flow cytometry, so that the glucose intake is obtained.
4. Glucose consumption experiment
The experimental groups were divided into 6 groups with 6 duplicate wells per group, and cells of each dosing group were incubated in a cell incubator for 24h. After the incubation, the absorbance (OD) values of each group of cell culture media were measured in a microplate reader at 505nm wavelength using a glucose assay kit (glucose oxidase method).
3. Experimental results
1. Effect of delta-juniper on HepG2 cell viability the cell viability of HepG2 cells incubated with 0-200 μm delta-juniper was not significantly changed, as shown in table 1, figure 1. Thus, the present study determined that the delta-juniper dosing concentrations were 2.5, 10, 17.5 μm, respectively.
TABLE 1 determination of the Activity of delta-juniper (0-200. Mu.M) on HepG2 cells
2. Effect of delta-juniperazine on glucose uptake by IR-HepG2 cells
Compared with the CON group, the MOD group cells have obviously reduced glucose uptake, the difference has statistical significance (P is less than 0.05), and the insulin sensitivity is reduced, so that insulin resistance is shown to occur, and the modeling is successful. While delta-juniper and ROSI groups at different concentrations increased glucose uptake by IR-HepG2 cells, the differences were statistically significant (P < 0.05), as shown in table 2, figure 2.
TABLE 2 Effect of different concentrations of delta-juniper on glucose uptake by IR-HepG2 cells
3. Effect of delta-juniperazine on glucose consumption in IR-HepG2 cells
The MOD cells showed a statistically significant decrease in glucose consumption (P < 0.01) compared to the CON cells, while the IR-HepG2 cells showed an improvement in glucose consumption after treatment with delta-juniper at different concentrations (P < 0.05), as shown in Table 3 and FIG. 3.
TABLE 3 influence of delta-juniper on the glucose consumption of IR-HepG2 cells
4. Conclusion(s)
In summary, the present study explored the role of delta-juniper in improving IR and sugar metabolism by constructing an IR-HepG2 cell model. The results show that after IR-HepG2 cells are incubated with delta-juniper in different concentrations, the glucose intake and consumption are obviously improved, the balance of glucose inside and outside the cells is maintained, and the effect that one of four southernwood is a nootropic monomer delta-juniper can play a role in reducing blood sugar by improving the effect of IR is proved.
The foregoing disclosure is merely illustrative of the preferred embodiments of the present invention and is not intended to limit the scope of the claims herein, as equivalent variations of the claims herein will fall within the scope of the invention.
Claims (7)
1. An application of a nootropic monomer delta-juniper in preparing a diabetes medicine is characterized in that: the intelligent monomer delta-juniper can be applied to the preparation of medicines for treating and/or preventing diabetes related diseases.
2. Use of the nootropic monomer delta-juniper according to claim 1 for the preparation of a medicament for diabetes, characterized in that: the dose concentration of the intelligent monomer delta-juniper alkene is 1-200 mu M.
3. Use of the nootropic monomer delta-juniper according to claim 2 for the preparation of a medicament for diabetes, characterized in that: the dose concentration of the intelligent monomer delta-juniper alkene is 1-100 mu M.
4. Use of the nootropic monomer delta-juniper in the manufacture of a medicament for diabetes according to claim 3, characterized in that: the dose concentration of the intelligent monomer delta-juniper alkene is 1-50 mu M.
5. The use of the nootropic monomer delta-juniper in the manufacture of a medicament for diabetes according to claim 4, characterized in that: the concentration of the nootropic monomer delta-juniper alkene administered was 2.5 mu M.
6. The use of the nootropic monomer delta-juniper in the manufacture of a medicament for diabetes according to claim 4, characterized in that: the intelligent monomer delta-juniper alkene administration concentration is 10 mu M.
7. The use of the nootropic monomer delta-juniper in the manufacture of a medicament for diabetes according to claim 4, characterized in that: the nootropic monomer delta-juniper is administered at a concentration of 17.5 μm.
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