CN114903886A - Application of tectorigenin in preparing medicine for preventing and treating diabetic cardiomyopathy - Google Patents
Application of tectorigenin in preparing medicine for preventing and treating diabetic cardiomyopathy Download PDFInfo
- Publication number
- CN114903886A CN114903886A CN202210601815.XA CN202210601815A CN114903886A CN 114903886 A CN114903886 A CN 114903886A CN 202210601815 A CN202210601815 A CN 202210601815A CN 114903886 A CN114903886 A CN 114903886A
- Authority
- CN
- China
- Prior art keywords
- tectorigenin
- diabetic cardiomyopathy
- artificial sequence
- preventing
- mice
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
Abstract
The invention provides application of tectorigenin in preparing a medicament for preventing and treating diabetic cardiomyopathy. According to the invention, through researching the influence of tectorigenin on a diabetic cardiomyopathy model mouse, the tectorigenin is confirmed to be capable of reducing hyperglycemia and hyperinsulinemia and cardiac dysfunction caused by the hyperglycemia and the hyperinsulinemia, wherein the cardiac dysfunction comprises the obvious reduction of fasting glycemia insulin, the obvious improvement of glucose tolerance and insulin, the obvious change of pathology and the improvement of cardiac function such as reversing left ventricular ejection fraction, and the like, and the tectorigenin can be used for preparing the medicine for preventing and treating diabetic cardiomyopathy.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to application of tectorigenin in preparing a medicine for preventing and treating diabetic cardiomyopathy.
Background
Diabetic cardiomyopathy is a special form of heart disease, a cardiac disease that is facilitated by resistance to insulin metabolism in heart tissue (e.g., insulin resistance), compensatory hyperinsulinemia, and the progression of hyperglycemia, occurring independently of other cardiac risk factors, such as Coronary Artery Disease (CAD) and hypertension. Clinical studies have confirmed that diabetic patients develop heart failure symptoms, while framinm et al have confirmed that men and women with diabetes have a 2.4-fold increased risk of heart failure as compared to individuals without diabetes, characterized by elevated left ventricular end-diastolic pressure, reduced left ventricular compliance, and low left ventricular ejection fraction with diffuse motor hypofunction. In this case, the diabetic's cardiac function eventually progresses to heart failure with a decrease in ejection fraction as the condition progresses. Therefore, the medicine is particularly important for the targeted treatment of diabetic cardiomyopathy, currently, medicines for reducing blood sugar such as metformin and dapagliflozin aiming at diabetic patients are mostly adopted for the medicine treatment of the diseases, the adverse reactions of the medicines are more, and the curative effect on the diabetic cardiomyopathy is not obvious, so that the medicine with small side effect is very important for searching for the medicine which can reduce blood sugar, improve insulin resistance and improve cardiac dysfunction.
Therefore, there is a need for the development of drugs for the prevention and treatment of diabetic cardiomyopathy.
Disclosure of Invention
The invention aims to provide application of tectorigenin in preparing a medicament for preventing and treating diabetic cardiomyopathy, wherein the tectorigenin can reduce hyperglycemia and hyperinsulinemia and cardiac dysfunction caused by the hyperglycemia and the hyperinsulinemia, such as obvious reduction of fasting glycemia and insulin, obvious improvement of glucose tolerance and insulin, obvious change of pathology and improvement of cardiac function such as reversal of left ventricular ejection fraction, and the like, and the application of the tectorigenin in preparing the medicament for preventing and treating diabetic cardiomyopathy is indicated.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides application of tectorigenin in preparing a medicament for preventing and treating diabetic cardiomyopathy. Including lowering blood sugar, improving insulin resistance and preventing and treating cardiac dysfunction caused by hyperglycemia and hyperinsulinemia.
The tectorigenin comprises a forming preparation containing active ingredients of tectorigenin, and a pharmaceutical composition synthesized by the tectorigenin and a pharmaceutically acceptable carrier or diluent.
The molding preparation comprises one of powder, tablets, granules, pills, capsules, oral liquid and injection.
The final concentration of tectorigenin is 5-100 μ g/ml.
Tectorigenin has molecular weight of 300.26 and molecular formula of C 16 H 12 O 6 The structure is shown as the following formula.
Because the tectorigenin has difference in pharmacological action dosage in various diseases, the invention also detects the safe dosage of the tectorigenin in the anti-diabetic cardiomyopathy, and mainly detects the influence of the tectorigenin on the vitality and the survival rate of the in vitro conventional culture myocardial cells and the safe dosage of an in vitro high-sugar induced diabetic cardiomyopathy model. Experiments prove that the tectorigenin with the concentration of 5-100 mu g/ml has no obvious drug toxicity effect on in vitro cultured myocardial cells and can inhibit pathological change of the myocardial cells stimulated by high sugar (HG, 33mmol/L), so that the tectorigenin has a full dose range for resisting the diabetes cardiomyopathy.
The invention proves that in a mouse experiment of tectorigenin for resisting diabetes cardiomyopathy, the weight of 50mg/kg of tectorigenin is administrated by intragastric administration to a mouse.
The invention mainly uses male C57/BL6 mice as experimental objects to confirm the application of tectorigenin in preparing the medicine for preventing and treating diabetic cardiomyopathy, and the specific experimental scheme is that a mouse model for treating diabetic cardiomyopathy is constructed by combining high-fat diet and Streptozotocin (STZ), a high-fat diet is firstly given for 3 months, then STZ (35mg/kg) is continuously injected for 5 days, fasting blood glucose measurement is carried out after one week, fasting blood glucose is more than or equal to 16.6mmol/L and is taken as the standard for successful construction of the model for treating diabetic cardiomyopathy, then grouping administration is carried out for 8 weeks, ultrasound and hemodynamic detection are carried out at 9 weeks of administration, and the left ventricle is taken to carry out detection of pathology and meristem level respectively so as to verify the application of tectorigenin in preparing the medicine for preventing and treating diabetic cardiomyopathy. The result proves that the tectorigenin can obviously inhibit cardiac function deterioration caused by hyperglycemia and hyperinsulinemia, and the expression is as follows: 1) the mice in the DCM + drug group had a reduced heart/body weight ratio compared to the diabetic cardiomyopathy group (DCM), and the administration group also improved the weight loss associated with diabetes; 2) fasting blood glucose and insulin levels of the administration group are both obviously reduced, and meanwhile, sugar tolerance (GTT) and insulin sensitivity (ITT) experiments are verified, so that the administration group can obviously improve glucose intolerance and insulin resistance of diabetic cardiomyopathy mice; 3) the ultrasonic results show that the cardiac function of the mice with diabetic cardiomyopathy is obviously improved; 4) pathological hematoxylin and eosin HE, PSR and Masson staining all showed improvement in myocardial and interstitial remodeling. In conclusion, tectorigenin can relieve cardiac dysfunction caused by hyperglycemia and hyperinsulinemia, and can be used for preparing medicines for preventing and treating diabetic cardiomyopathy.
One or more technical solutions in the embodiments of the present invention at least have the following technical effects or advantages:
the invention provides application of tectorigenin in preparation of a medicament for preventing and treating diabetic cardiomyopathy, which confirms that the tectorigenin can reduce hyperglycemia and hyperinsulinemia and cardiac dysfunction caused by the hyperglycemia and the hyperinsulinemia through researching the influence of the tectorigenin on a diabetic cardiomyopathy model mouse, wherein the cardiac dysfunction comprises the improvement of obviously reducing fasting glycemia and insulin, obviously changing pathology, reversing left ventricular ejection fraction and the like, and the application indicates that the tectorigenin is used for preparing the medicament for preventing and treating diabetic cardiomyopathy.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the description below are some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on the drawings without creative efforts.
FIG. 1 is a graph showing the absorbance values at 450nm of each dose group measured by purchasing a CCK-8 kit for determining the optimum drug dose of irigenin, and further analyzing the cell viability to determine the optimum non-toxic dose of irigenin according to example 1.
FIG. 2 is the change in body weight of the mice during feeding in example 1. FIGS. 2A-D show changes in body weight, heart weight, and food intake of mice.
FIG. 3 is a graph of the effect of the drug on mouse glucose tolerance and insulin sensitivity in example 3. FIGS. 3A-C are the results of fasting plasma glucose and insulin in mice and steady state model assessment of insulin resistance (HOMA-IR); FIGS. 3D-G are graphs of sugar tolerance GTT experiments and insulin resistance ITT experiments.
FIG. 4 shows the results of the in vivo cardiac function test in example 4. FIGS. 4A-D are statistics of left ventricular ejection fraction, short axis shortening rate, left ventricular end-diastolic diameter, and diastolic ventricular septal thickness, respectively; FIGS. 4E-F are hemodynamic pressure-volume measurements.
Figure 5 is a pathological staining and associated index detection map of example 5. FIGS. 4A-D are graphs of sirius red (PSR), Masson stain, and overall collagen content and collagen area; FIGS. 5E-F are the pathological results hematoxylin-eosin (HE) staining and cardiomyocyte area measurements.
FIG. 6 is the level of change of the central function-associated molecular marker of example 6. FIGS. 6A-C are in vivo transcript levels of ANP, BNP, and β -MHC; FIGS. 6D-G show the in vivo transcript levels of α -SMA, CTGF, COL-1, and COL-3.
FIG. 7 is an immunofluorescent staining and associated index detection map of high glucose stimulated isolated cardiomyocytes. FIG. 7A shows immunofluorescence and cardiomyocyte area measurements; FIGS. 7B-E are ex vivo transcript levels of ANP, BNP, and β -MHC; FIG. 7F is an immunofluorescence staining pattern for α -SMA.
Detailed Description
The present invention will be described in detail below with reference to specific embodiments and examples, and the advantages and various effects of the present invention will be more clearly apparent therefrom. It will be understood by those skilled in the art that these specific embodiments and examples are for the purpose of illustrating the invention and are not to be construed as limiting the invention.
Throughout the specification, unless otherwise specifically noted, terms used herein should be understood as having meanings as commonly used in the art. Accordingly, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is a conflict, the present specification will control.
Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be obtained by an existing method.
Isolated cardiomyocyte culture: the experiment mainly adopts H9C2 cells, and DMEM medium containing 20% fetal calf serum is placed at 37 ℃ in 5% CO 2 In the incubator, the culture is performed. After the cells are full to about 80 percent, digesting with 0.125 percent of pancreatin and passaging the cells according to the proportion of 10 4 The density of/ml was seeded in 6-well or 24-well plates for subsequent experiments.
Feeding experimental animals: the invention mainly adopts C57/BL6 with the age of 8 weeks, is purchased from the institute of medical laboratory animals of Chinese academy of science and medicine, and is raised in the center of laboratory animals of the institute of cardiovascular diseases of Wuhan university.
The source of the drug is as follows: the medicine used in the invention is purchased from Shanghai Huo medical science and technology development Limited company, with the number of 548-77-6, and is powder with the purity of more than 98%.
Preparing the medicine: in vitro cell assay configuration: dissolving tectorigenin with 0.1% dimethyl sulfoxide (DMSO, Sigma # D2650) to prepare 20mM stock solution, storing in a refrigerator at-20 ℃, diluting the medicament to the prior safe dose during the experiment so that the final concentration of the medicament is 5-100 mu g/ml, and carrying out in vitro experiment; in vivo mouse gavage drug configuration: tectorigenin was dissolved in 0.1% DMSO in physiological saline to prepare a drug concentration of 5 mg/ml.
The administration mode comprises the following steps: the prepared diabetic cardiomyopathy model mouse is subjected to intragastric administration according to the weight of 50mg/kg, the medicine is used as it is, and the administration is started 1 time/day after one week of STZ injection and lasts for 8 weeks.
Example 1 toxicity assay of tectorigenin on isolated cardiomyocytes
1. H9C2 cardiomyocyte culture
Culturing H9C2 cells which have been passaged for 3-5 times and have good state in a DMEM medium containing 20% fetal calf serum, and placing the cells at 37 ℃ in the DMEM medium 2 The culture box is digested by pancreatin and inoculated in a six-well plate or a 24-well plate for carrying out corresponding cell experiments.
2. Myocardial cell dosing and counting
1) Grouping: the total groups were 7, including mainly blank control group: medium containing only the same dose is cell free; control group without drug: the group of cells was not stimulated by tectorigenin; different dosages of tectorigenin, such as 20. mu.g/mL, 30. mu.g/mL, 50. mu.g/mL, 100. mu.g/mL, 200. mu.g/mL, 300. mu.g/mL, 400. mu.g/mL, were added to the cells and incubated with the control group for 24 hours.
2) Cell activity assay: after administration of the corresponding drug dose of H9C2 cells seeded in a 96-well plate, 10. mu.l of CCK-8 solution was added to each well, and after incubation in an incubator for 2 hours, absorbance (A) at 450nm was measured with a microplate reader, and cardiomyocyte activity was calculated as follows: cell activity (%) ═ treatment group a/control group a × 100%.
FIG. 1A shows the results: compared with a control group, the tectorigenin dose has no significant influence on the myocardial cell activity when being 20 mu g/mL, 30 mu g/mL, 50 mu g/mL, 100 mu g/mL, 200 mu g/mL, 300 mu g/mL and 400 mu g/mL, and the myocardial cell activity is significantly reduced when the tectorigenin dose is 200 mu g/mL or more, and the results show that the tectorigenin dose has no obvious toxic effect on the myocardial cell when being 5-100 mu g/mL and is a safe dose range of the tectorigenin.
3. High glucose post stimulation of myocardial cells dosing and counting
1) Grouping: the method mainly comprises six groups, namely, a blank control group: culturing the cells in cell culture medium for 24h without any stimulation; high sugar stimulation group: culturing the cells in a cell culture medium for 24h, adding high sugar to make the final concentration 33Mm, and stimulating for 24 h; high carbohydrate stimulation + tectorigenin in different dose groups, using doses within the above-identified safe range of tectoridin doses, including four doses of 25. mu.g/mL, 50. mu.g/mL, 100. mu.g/mL, 200. mu.g/mL.
2) Cell activity assay: as before. FIG. 1B shows the results: after stimulation with high sugar, the effect on myocardial cell protection was most pronounced when the irigenin dose was 50 μ g/mL compared to the control. Thus, this example demonstrates that the optimal concentration of irigenin applied to ex vivo H9C2 cardiomyocytes after high glucose stimulation is 50 μ g/mL.
Example 2 tectorigenin changes in body weight and Heart weight in mice
1. Establishing mouse diabetic cardiomyopathy model and tectorigenin intervention
Four groups of C57/BL6 mice were designed for this experiment, including: (ii) control group (Con); ② tectoridin (Tec); ③ diabetic cardiomyopathy group (DCM); diabetic cardiomyopathy + tectoridin (DCM + Tec).
Mice of 8-week age are randomly divided into two groups, namely a control group and a model building group, the model building group is fed with high fat for 12 weeks, STZ5 is continuously injected in the 13 th week, then adaptive feeding is carried out for 1 week, fasting blood glucose is measured, the mice with the blood glucose concentration meeting the standard are brought into the experiment, and the mice are randomly divided into iriside groups, diabetic cardiomyopathy and iriside groups.
After the components are separated, the stomach irrigation administration is started, the tectorigenin is dissolved in 0.1 percent DMSO normal saline, and the final concentration of the tectorigenin solution is 5 mg/mL; tectoridin and diabetic cardiomyopathy + tectoridin mice were dosed at 50mg/kg body weight, 0.2 ml/dose, 1 dose/day, for 8 weeks.
2. The body weight of the mice was measured weekly since the administration, and the growth rate was compared and counted for 8 weeks.
3. Food intake measurements were recorded weekly.
4. Mouse heart tissue was taken and evaluated at the histological level for myocardial and interstitial remodeling.
5. Weighing the weight of the mice after 8 weeks of administration, killing the mice, taking the materials, opening the chest of the mice after death immediately, taking out the hearts which are not stopped, quickly putting the hearts into a 10% KCl solution, keeping the hearts in the diastole, trimming the rest involved tissues, gently pressing the tissues to drain the extravasated blood, and weighing (mg); lung tissue was removed for weight recording (mg); one side of the tibia was exposed and its length (cm) was measured. And (3) completely putting the weighed heart of the mouse into formalin for fixation for 24-48 hours, then trimming and putting the heart into an embedding frame for dehydration, transparency and embedding, and preparing pathological sections for subsequent experiments.
Results figures 2A-B show that tectorigenin has a remitting effect on weight loss caused by diabetes, during which there was no significant difference in food intake; FIGS. 2D-C show that tectorigenin is resistant to hyperinsulinemic induced weight gain (P < 0.05);
example 3 Effect of irigenin on mouse glucose tolerance and insulin resistance
Glucose Tolerance Tests (GTTs) and Insulin Tolerance Tests (ITTs) were performed separately in mice treated with tectorigenin for 8 weeks. Briefly, mice were fasted overnight, then were injected intraperitoneally with glucose for the GTT assay or with insulin for the ITT assay. Glucose levels were then determined at 0, 15, 30, 60 and 120 minutes post intravenous injection. Specifically, a Roche glucometer and mouse tail vein blood sampling are adopted for blood sugar measurement.
The results in FIGS. 3A-B show that irigenin can lower blood glucose and hyperinsulinemia levels (P <0.05) in diabetic cardiomyopathy mice;
figures 3C-G show that tectorigenin can improve glucose intolerance and insulin resistance (P <0.05) in diabetic cardiomyopathy mice.
Example 4 Effect of tectorigenin on cardiac function in diabetic cardiomyopathy mice
1. Establishing mouse cardiac muscle reconstruction model and tectorigenin intervention
The contents are the same as in example 2.
2. Echocardiography for evaluating cardiac function of mice
Firstly, setting various basic parameters of an ultrasonic instrument; grabbing the mouse for weight measurement and recording, removing the hair of the precordial region of the mouse, carrying out anesthesia by using 1.5-2.5% isoflurane, smearing an ultrasonic coupling agent when the mouse enters an anesthesia state, horizontally placing the mouse in a lying position, detecting and recording the heart short axis shortening rate, the left ventricle ejection fraction, the left ventricle diastolic end internal diameter and the diastolic interval thickness of the mouse in the left ventricle papillary muscle short axis section level.
3. Hemodynamics detection and evaluation of mouse cardiac function
Hemodynamics were tested by inserting a Millar 1.4F microcatheter into the left ventricle along the common carotid artery. Pressure signals and heart rate were continuously recorded using a Millar pressure volume catheter (SPR-839; Millar Instruments) connected to a Powerlab system (AD Instruments Ltd., UK), the resulting data and related images were stored to a computer, and finally the collected data was analyzed using PVAN data analysis software.
FIG. 4 shows the results: the left ventricular end-diastolic inner diameter and diastolic ventricular septal thickness of the diabetic cardiomyopathy group were increased compared to the control group, but both the left ventricular ejection fraction and the short axis shortening rate of the mice were decreased (P < 0.05); tectorigenin treated groups restored increased left ventricular end-diastolic inner diameter and diastolic ventricular septal thickness in diabetic cardiomyopathy mice and improved reductions in ejection fraction and short axis shortening (P <0.05) (fig. 4A-C). FIGS. 4D-E show the results: compared with the control group, the maximal rising rate (dp/dtmax) and the falling rate (dp/dtmin) of the left ventricular pressure of the mice in the diabetic cardiomyopathy group are both reduced (P is less than 0.05), and the hemodynamics of the mice are prompted to be in disorder; and the tectorigenin treatment group improves left ventricular compliance of diabetic cardiomyopathy mice, and reduces dp/dtmax and dp/dtmin (P <0.05) of the diabetic cardiomyopathy resistant mice. The diabetic cardiomyopathy is characterized by decreased ventricular wall compliance, hemodynamic disturbance and the like, changes of cardiac functions are evaluated by ultrasonic and hemodynamic detection, and the tectorigenin is found to improve the cardiac dysfunction of diabetic cardiomyopathy mice and is mainly characterized by improvement of ventricular wall compliance and hemodynamic.
Example 5 Effect of tectorigenin on Heart pathologies and related indices
1. Heart tissue from mice was collected and evaluated at the histological level for the degree of cardiac dysfunction
The contents are the same as in example 2.
2. Sirius red PSR staining: baking the 5-micron paraffin section in a 65-degree oven for more than 30min to enable the tissue section to be tightly attached to the glass slide; carrying out xylene dewaxing and 100-70% gradient alcohol hydration; the trypsin method antigen repairing section, PBS rinse will slice, will slice put into 0.2% phosphomolybdic acid for 1-5 min, 0.1% picric acid Tianlang red staining solution drop on the tissue, then will slice put into a moist dyeing box to dye for 1.5h, later in 0.01N hydrochloric acid under 4, 70% -100% gradient alcohol dehydration, xylene transparent, finally use the neutral resin to seal the piece, will be placed in the fume hood and dried, can take a picture. And (4) dyeing and photographing: cells with at least 6 well-defined nuclei in the center of the cells were required for each Image, and the collagen area was measured using Image-Pro Plus 6.0 Image analysis software after photographing.
(1) Masson staining: 1) dewaxing: baking the slices at 65 ℃ for more than 30min, simultaneously preheating the bouin stationary liquid, and performing xylene dewaxing and 100-70% gradient alcohol hydration; 2) fixing: adding the bouin stationary liquid dropwise, and heating in an incubator at 37 ℃ for 1 h; 3) ddH2O washed until the yellow color disappeared; 4) hematoxylin staining for 5min → Lichun fuchsin 5-10min → pure water washing in a jar until the water is colorless → 2% glacial acetic acid is dripped and dyed for 3-5min → 5.1% phosphomolybdic acid is differentiated for 3-5min, 6.2% aniline blue is washed in a jar until the water is colorless → 0.2% glacial acetic acid is dripped and dyed for 5min → 70% -100% gradient alcohol is dehydrated, the xylene is transparent, and finally, a neutral resin is used for sealing the piece, and the piece is placed in a ventilation drying cabinet for shooting.
(2) Hematoxylin-eosin (HE) staining for myocardial cell area: baking slices, dewaxing and hydrating; then, performing hematoxylin staining solution, 1% hydrochloric acid alcohol differentiation, performing Scott solution bluing, and rinsing with eosin staining solution and distilled water; mounting and observing under microscope. HE staining photograph: the requirements are as before.
The results of FIG. 5 show: compared with the control group, the total volume of the heart in the diabetic cardiomyopathy group is increased, the cross sectional area of myocardial cells is increased, the arrangement of myocardial interstitium is disordered, the collagen content is increased, interstitial obvious edema is caused, and a large amount of inflammatory cell infiltration (P <0.05) can be seen; tectorigenin can reduce total volume of heart and cross-sectional area of myocardial cells, and relieve interstitial edema, inflammatory cell infiltration and myocardial interstitial arrangement disorder degree (P < 0.05). The results prove that the tectorigenin can obviously relieve myocardial and interstitial reconstruction caused by high glucose and high insulin.
Example 6 Effect of tectorigenin on meristematic indicators associated with in vivo cardiac function
1. RT-PCR detection of mouse in vivo myocardial lesion related index transcription level
After 8 weeks of dosing, mice were weighed, sacrificed and the material was harvested, the heart was squeezed clean of blood and trimmed and weighed (mg); grinding heart tissues of mice to extract total RNA, and detecting the in vivo transcription level of ANP, BNP, beta-MHC, alpha-SMA, CTGF, COL-1 and COL-3. The specific method adopts TRIzol to extract total RNA of the myocardial cells, carries out reverse transcription to obtain cDNA, and utilizes a Light-Cycler 480SYBR instrument to detect. The primer sequences are as follows:
TABLE 1
The results of FIG. 6 show: compared with a control group, the transcriptional levels of ANP, BNP, beta-MHC, alpha-SMA, CTGF, COL-1 and COL-3 in the diabetic cardiomyopathy group are all increased, and the difference has statistical significance (P is less than 0.05); while tectorigenin can inhibit the increased transcription level of the myocardial damage marker in diabetic cardiomyopathy groups (P < 0.05). The result of the damage of myocardial cells caused by hyperglycemia and hyperinsulinemia indicates that the damage of the myocardial cells caused by hyperglycemia and hyperinsulinemia can be reversed.
Example 7 detection of the Effect of tectorigenin on isolated hyperglycosylated cardiomyocytes
1. H9C2 cardiomyocyte culture
The contents are the same as in example 2.
2. High-sugar stimulation of cardiomyocytes and intervention with tectorigenin
Grouping (Con) according to experimental requirements; ② tectoridin (Tec); high glucose stimulation group (HG); high sugar stimulation and tectoridin (HG + Tec) wherein the high sugar stimulation concentration is 33mmol/L, and the tectorigenin concentration is 50 mu g/mL.
3. H9C2 cell immunofluorescent staining to observe cell changes
Taking out the 24-well plate with the cells from the incubator, washing the 24-well plate with PBS for 3 times, and fixing the plate with 4% paraformaldehyde for 10min at room temperature; 0.2% Triton permeabilized cardiomyocytes; sealing the goat serum for 1 h; alpha-actin (TACCAM, TAC68167) and 1% sheep serum were diluted 1:100 to formulate primary antibody and overnight at 4 ℃. Alexa Fluor 488 secondary antibody (secondary antibody: PBS 1: 200) was diluted with PBS daily, dropped on a slide, incubated at 37 ℃ for 1h, rinsed with PBS three times, dropped with DAPI to develop nuclei, and photographed by observation under an upright microscope.
The results of FIGS. 7A-B show: high sugars can induce changes in cardiomyocyte volume and collagen content, as evidenced by increased cardiomyocyte cross-sectional area (P < 0.05); the addition of tectorigenin can obviously inhibit the pathological change of myocardial cells (P < 0.05).
4. Marker for detecting hypertrophy of isolated cardiac muscle cells by RT-PCR
After the corresponding stimulation of H9C2 cardiomyocytes seeded in six-well plates, total RNA of cardiomyocytes was extracted using TRIzol, reverse-transcribed to cDNA, and the transcription levels of hypertrophy markers such as Atrial Natriuretic Peptide (ANP), Brain Natriuretic Peptide (BNP), and β -type myosin heavy chain (β -MHC) in cardiomyocytes were detected using Light-Cycler 480SYBR instrument. The primer sequences are as follows:
TABLE 2
The results of FIGS. 7C-D show: compared with the control group without the drug, the mRNA expressions of ANP, BNP and beta-MHC in the hyperglycemia group are all increased, and the difference has statistical significance (P is less than 0.05); and the transcriptional level of the cardiac muscle damage marker of the tectoridin group is obviously reduced compared with that of the high-sugar stimulation group, and the difference has statistical significance (P < 0.05). Thus, this example partially defines that tectorigenin has protective effects on isolated cardiomyocytes after high-sugar stimulation.
Conclusion
In conclusion, the medicament tectorigenin has obvious inhibition effect on hyperglycemia and hyperinsulinemia of diabetic cardiomyopathy, can relieve cardiac dysfunction caused by hyperglycemia and hyperinsulinemia, and can be obtained by pathological staining and in-vivo and in-vitro myocardial injury related molecular index detection.
Finally, it should also be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Sequence listing
<110> Wuhan university
Application of tectorigenin in preparation of medicine for preventing and treating diabetic cardiomyopathy
<160> 24
<170> SIPOSequenceListing 1.0
<210> 1
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
tggctctgtg caagaagcgg at 22
<210> 2
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
aatctgggcg ttgtcactgc gt 22
<210> 3
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
<210> 4
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 4
gaagcacagg agcaggtgta ga 22
<210> 5
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 5
<210> 6
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 6
gaagcacagg agcaggtgta ga 22
<210> 7
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 7
atcctgtcca aactaaggct cg 22
<210> 8
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 8
acctctttag catagtagtc cgc 23
<210> 9
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 9
<210> 10
<211> 25
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 10
ccttggctgt tatcttcggt accgg 25
<210> 11
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 11
gaggtcactc ctatcctctg g 21
<210> 12
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 12
gccatttcct ccgacttttc tc 22
<210> 13
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 13
ccgagtccca ggtcaacaa 19
<210> 14
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 14
cttcacgggc acccttgga 19
<210> 15
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 15
actccactca cggcaaattc 20
<210> 16
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 16
<210> 17
<211> 24
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 17
aaagcaaact gagggctctg ctcg 24
<210> 18
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 18
ttcggtaccg gaagctgttg ca 22
<210> 19
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 19
acaatccacg atgcagaagc t 21
<210> 20
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 20
gggccttggt cctttgaga 19
<210> 21
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 21
<210> 22
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 22
caaggctaac ctggagaaga tg 22
<210> 23
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 23
<210> 24
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 24
Claims (4)
1. Application of tectorigenin in preparing medicine for preventing and treating diabetic cardiomyopathy is provided.
2. The use according to claim 1, wherein the tectorigenin comprises a shaped formulation comprising an active ingredient of tectorigenin, and the tectorigenin is combined with a pharmaceutically acceptable carrier or diluent to form a pharmaceutical composition.
3. The use of claim 1, wherein the shaped form comprises one of a powder, a tablet, a granule, a pill, a capsule, an oral liquid, and an injection.
4. The use according to claim 1, wherein tectorigenin is administered to a final concentration of 5 to 100 μ g/ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210601815.XA CN114903886A (en) | 2022-05-30 | 2022-05-30 | Application of tectorigenin in preparing medicine for preventing and treating diabetic cardiomyopathy |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210601815.XA CN114903886A (en) | 2022-05-30 | 2022-05-30 | Application of tectorigenin in preparing medicine for preventing and treating diabetic cardiomyopathy |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114903886A true CN114903886A (en) | 2022-08-16 |
Family
ID=82770826
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210601815.XA Pending CN114903886A (en) | 2022-05-30 | 2022-05-30 | Application of tectorigenin in preparing medicine for preventing and treating diabetic cardiomyopathy |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114903886A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE10123503A1 (en) * | 2001-05-15 | 2002-11-21 | Bionorica Ag | Use of Belamcanda chinensis extract, or its active ingredient, for treating sex hormone-dependent urogenital disorders and prostatic hyperplasia |
| CN109078011A (en) * | 2018-10-01 | 2018-12-25 | 东北师范大学 | The application of iris aglycone and its derivative in prevention and treatment insulin resistance disease medicament |
| CN109549938A (en) * | 2018-10-22 | 2019-04-02 | 烟台大学 | Purposes of the procyanidin compounds in the product for preventing and/or treating insulin resistance |
| CN113304149A (en) * | 2021-06-23 | 2021-08-27 | 广州医科大学 | Application of compound in preparation of medicine for treating type 2 diabetic cardiomyopathy |
-
2022
- 2022-05-30 CN CN202210601815.XA patent/CN114903886A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE10123503A1 (en) * | 2001-05-15 | 2002-11-21 | Bionorica Ag | Use of Belamcanda chinensis extract, or its active ingredient, for treating sex hormone-dependent urogenital disorders and prostatic hyperplasia |
| CN109078011A (en) * | 2018-10-01 | 2018-12-25 | 东北师范大学 | The application of iris aglycone and its derivative in prevention and treatment insulin resistance disease medicament |
| CN109549938A (en) * | 2018-10-22 | 2019-04-02 | 烟台大学 | Purposes of the procyanidin compounds in the product for preventing and/or treating insulin resistance |
| CN113304149A (en) * | 2021-06-23 | 2021-08-27 | 广州医科大学 | Application of compound in preparation of medicine for treating type 2 diabetic cardiomyopathy |
Non-Patent Citations (4)
| Title |
|---|
| KYUNG-TAE LEE 等: "Hypoglycemic and Hypolipidemic Effects of Tectorigenin and Kaikasaponin III in the Streptozotocin-lnduced Diabetic Rat and their Antioxidant Activity In vitro", ARCH PHARM RES, vol. 23, no. 5, pages 461 - 466 * |
| SANG HOON JUNG 等: "Isoflavonoids from the Rhizomes of Belamcanda chinensis and Their Effects on Aldose Reductase and Sorbitol Accumulation in Streptozotocin Induced Diabetic Rat Tissues", ARCH PHARM RES, vol. 25, no. 3, pages 306 - 312, XP053006897, DOI: 10.1007/BF02976631 * |
| 李增玉 等: "醛糖还原酶与炎症性疾病", 中国当代医药, vol. 19, no. 16, pages 17 - 18 * |
| 王金凤 等: "鸢尾苷元胃内漂浮缓释片兔体内药动学及其体内外相关性研究", 中草药, vol. 48, no. 2, pages 266 - 271 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20160129077A1 (en) | Use of recombinant ganoderma immunoregulatory protein (rLZ-8) in preparation of drug for treating melanoma | |
| CN117323323A (en) | Activation of AMPK by Phoebe cheilis component | |
| CN108434127B (en) | Application of myricanol and/or myricetin in preparation of medicine for preventing and/or treating inflammatory bowel disease | |
| JP6180417B2 (en) | Use of compounds for the manufacture of therapeutic agents for heart failure or neuronal damage | |
| KR20180100152A (en) | Use of triacetyl-3-hydroxyphenyl adenosine in the manufacture of a medicament for the treatment or prevention of nonalcoholic fatty liver disease | |
| CN114903886A (en) | Application of tectorigenin in preparing medicine for preventing and treating diabetic cardiomyopathy | |
| CN110038002B (en) | Application of salvianolic acid A in preventing and treating muscular atrophy, myopathy and musculoskeletal complications | |
| CN107157980B (en) | Application of oridonin in preparation of anti-myocardial remodeling drugs | |
| CN114209810B (en) | Application of lycium barbarum glycopeptide in preparation of medicine for preventing or treating inflammatory bowel disease | |
| CN103356630B (en) | Containing pentoxifylline and the pharmaceutical composition of prucalopride and medical usage thereof | |
| CN112107599B (en) | Application of stem cell exosome in preparation of medicine for treating overactive bladder syndrome | |
| CN114306304A (en) | Application of 4-hydroxybenzoic acid in preparing medicine for improving myocardial infarction complication and medicine | |
| CN103735550A (en) | Medicinal composition for treating CKD (chronic kidney disease) myocardial injury and application thereof | |
| CN106860449A (en) | Purposes of the matrine derivative in diabetes are treated | |
| CN114588212A (en) | New use of traditional Chinese medicine water lettuce for resisting heart failure | |
| CN114917220A (en) | Application of tectorigenin in preparing medicine for preventing and treating myocardial remodeling | |
| CN115227700B (en) | Application of esculentoside A in preparation of myocardial infarction protection medicine | |
| CN109820903A (en) | A kind of Chinese medicine composition and the preparation method and application thereof for treating diabetic nephropathy | |
| CN108653401A (en) | A kind of compound medicinal formulation and preparation method thereof for treating ulcerative colitis | |
| CN114288301B (en) | Application of DTQ in preparation of medicines for treating acute myocardial infarction and related products | |
| CN115364142B (en) | Application of Qihuangyi heart oral liquid in preparation of medicine for treating diabetic cardiomyopathy | |
| CN114712379B (en) | Application of astragaloside IV in preparing medicine for preventing and treating peritoneal dialysis intestinal complications | |
| CN110051671B (en) | Application of purslane amide E in preparation of medicine for treating ischemic heart disease | |
| CN111632059B (en) | New application of swertiamarin | |
| CN116421610A (en) | Application of wogonin in preparation of antihypertensive products |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |