A medicament for delaying vascular aging
By utilizing the synergistic effect of gentian bitter sugar and Ganoderma lucidum polysaccharides, drugs or culture media were prepared, which solved the problem of endothelial cell aging caused by angiotensin II and achieved the effect of delaying vascular aging.
Patent Information
- Application Number
- CN202310292916.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-03-23
- Publication Date
- 2025-11-21
- Estimated Expiration
- 2043-03-23
AI Technical Summary
Vascular aging is a major pathogenic mechanism leading to atherosclerosis, heart failure, and hypertension. Current technologies are insufficient to effectively reduce endothelial cell aging caused by angiotensin II.
Gentian root extract and Ganoderma lucidum polysaccharide were used as active ingredients to prepare liquid drugs or culture media at concentrations of 20 μmol/L and 100 mg/L, respectively, which synergistically reduced endothelial cell senescence induced by angiotensin II.
It significantly reduces angiotensin II-induced vascular endothelial cell senescence, decreases Cav-1 protein expression, and slows down the vascular aging process.
Smart Images
Figure CN116407550B_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of cardiovascular and cerebrovascular diseases, and particularly relates to a medicine for delaying blood vessel aging. BACKGROUND
[0002] Aging is an inevitable progressive decline in the function of physiological organs, which increases the probability of disease and death. Among them, blood vessel aging is an important pathological and physiological basis for the aging of human organs and tissues, and is also the main pathogenesis of many old-age diseases such as atherosclerosis, heart failure and hypertension. The cause of blood vessel aging is the change of clinical and subclinical results and functions of blood vessel wall cells and extracellular matrix at the tissue and cell level.
[0003] Angiotensin II (Ang II) is an inflammation trigger that activates inflammatory factors such as NF-KB and TGF-β1 in the blood vessel wall, thereby promoting the inflammatory response of the blood vessel, and can stimulate cells to produce free radicals, leading to the aging of vascular endothelial cells, and further leading to the occurrence and development of atherosclerosis. Therefore, reducing the aging of endothelial cells caused by angiotensin II will help to reduce blood vessel aging. SUMMARY
[0004] The purpose of the present application is to provide a medicine for delaying blood vessel aging by reducing angiotensin-induced endothelial cell aging.
[0005] To achieve the above-mentioned purpose, the present application provides the following technical solutions:
[0006] The present application provides a medicine for delaying blood vessel aging, which is a liquid medicine, and the medicine comprises gentiopicroside, ganoderma lucidum polysaccharide and a pharmaceutically acceptable carrier.
[0007] Preferably, the concentration of gentiopicroside in the medicine is 20 μmol / L, and the concentration of ganoderma lucidum polysaccharide in the medicine is 100 mg / L.
[0008] Preferably, the medicine is used for endothelial cell aging caused by angiotensin II.
[0009] Secondly, the present application provides a medicine for reducing blood vessel endothelial cell aging, which is a liquid medicine, and the medicine comprises gentiopicroside, ganoderma lucidum polysaccharide and a pharmaceutically acceptable carrier.
[0010] Preferably, the concentration of gentiopicroside in the medicine is 20 μmol / L, and the concentration of ganoderma lucidum polysaccharide in the medicine is 100 mg / L.
[0011] Secondly, the present application provides a culture medium for reducing blood vessel endothelial cell aging, which contains gentiopicroside and ganoderma lucidum polysaccharide.
[0012] Preferably, the concentration of gentiopicroside in the culture medium is 20 μmol / L, and the concentration of ganoderma polysaccharide in the culture medium is 100 mg / L.
[0013] Secondly, the present application provides the use of gentiopicroside in the preparation of a medicine for delaying blood vessel aging.
[0014] Preferably, the medicine is a liquid medicine, and the concentration of gentiopicroside in the medicine is 20 μmol / L.
[0015] In addition, the present application provides the use of gentiopicroside in the preparation of a culture medium for reducing the aging of vascular endothelial cells, and the concentration of gentiopicroside in the culture medium is 20 μmol / L.
[0016] The present application has the following beneficial effects:
[0017] Firstly, the present application finds that gentiopicroside can reduce the aging of vascular endothelial cells caused by angiotensin II and increase the expression of Cav-1 protein caused by angiotensin II.
[0018] Secondly, the present application finds that gentiopicroside can produce a synergistic effect with ganoderma polysaccharide to greatly reduce the aging of vascular endothelial cells caused by angiotensin II, so that the two can be used in combination to prepare an efficient medicine for delaying blood vessel aging. BRIEF DESCRIPTION OF DRAWINGS
[0019] Figure 1 Effect of gentiopicroside on the up-regulation of Cav-1 protein (caveolin) caused by angiotensin II. DETAILED DESCRIPTION
[0020] In order to clearly illustrate the technical features of the present application, the present application will be described below through specific embodiments.
[0021] Gentiopicroside is a natural iridoid glycoside, which is one of the main pharmacodynamic components of Gentiana plants, and widely exists in the dried roots and rhizomes of plants such as Gentiana scabra Bunge, and its structure is as follows:
[0022]
[0023] The present application mainly discusses whether gentiopicroside can play a role in the aging of vascular endothelial cells.
[0024] Example 1: Effect of gentiopicroside on the activity of human vein endothelial cells (HUVEC) 3 The HUVEC cells in the logarithmic growth phase were inoculated in a 96-well plate at 3×10
[0025] The control group and the gentian bitter-sweet group were set up (10, 20, 40 μmol / L). The gentian bitter-sweet group was given the drug according to the drug concentration, while the control group was given the culture medium without the drug. Each group had 3 replicate wells.
[0026] After culturing for 24 hours, add 10 μL of CCK-8 working solution to each well. After incubation, use a microplate reader to detect the absorbance at 450 nm.
[0027] Example 2: Effects of Gentian Bitter-Sweet on Angiotensin II-Induced Cellular Senescence
[0028] HUVEC cells were seeded in 6-well plates and treated according to the following experimental groups;
[0029] Experimental Groups:
[0030] Control group: Cultured in culture medium containing 10% serum, without the addition of any drugs;
[0031] Angiotensin II group: Add 10 -6 Angiotensin II at mol / L was used, and the solution was replaced after 24 hours, for a total of 48 hours.
[0032] Gentianoside group: After adding 20 μmol / L for 1 h, add 10 -6 Angiotensin II at mol / L was used, and the solution was replaced after 24 hours, for a total of 48 hours.
[0033] After the treatment, the cell culture medium was removed, the cells were washed with PBS, and then fixed with glutaraldehyde for 10 min.
[0034] After washing three times with PBS, the PBS was removed, and 1 ml of X-gal staining solution was added to each well. After incubation at 37°C for 12 h, five fields of view were randomly selected from each group under a fluorescence microscope, and the percentage of positive cells was calculated.
[0035] Example 3: Effects of gentian bitterness and licorice on angiotensin II-induced upregulation of Cav-1 protein (foveolar protein).
[0036] Angiotensin II leads to increased expression of Cav-1 protein, and high expression of Cav-1 protein activates cellular senescence signals, leading to cellular senescence. Therefore, this invention studies the effect of gentian bitter licorice on the upregulation of Cav-1 protein caused by angiotensin II.
[0037] Control group: Cultured in culture medium containing 10% serum, without the addition of any drugs;
[0038] Angiotensin II group: 10⁻⁶ mol / L angiotensin II was added, and the solution was replaced after 24 hours of treatment, for a total of 48 hours.
[0039] Gentiopicroside group: 20 μmol / L gentiopicroside was added for 1 h, then 10-6mol / L angiotensin II was added, and the medium was replaced after 24 h, and the treatment was performed for 48 h in total;
[0040] After the treatment, the cell culture medium was removed, the cells were washed with PBS, and protein lysate was added to extract the protein sample;
[0041] The concentration of the protein sample in each group was detected by referring to the BCA protein detection kit;
[0042] After adding the loading buffer, boiling water was heated for denaturation for 5 min, and then placed at -20°C for standby;
[0043] A 12% protein electrophoresis gel was prepared, and 20 μg of sample was loaded per well. After the protein sample was electrophoresed on the 12% gel, it was transferred to the PVDF membrane by electrophoresis;
[0044] The PVDF membrane was blocked with TBST containing 5% skim milk at room temperature for 1 h with slow shaking;
[0045] Cav-1 primary antibody and GAPDH primary antibody were used for incubation at 4°C overnight;
[0046] After TBST washing, the secondary antibody was incubated at room temperature for 1 h. After washing the membrane, chemiluminescence solution was used for color development.
[0047] Example 4: Effect of gentiopicroside and lentinan on cell senescence caused by angiotensin II
[0048] Control group: cultured with culture medium containing 10% serum without adding drugs;
[0049] Angiotensin II group: 10-6mol / L angiotensin II was added, and the medium was replaced after 24 h, and the treatment was performed for 48 h in total;
[0050] Gentiopicroside group: 20 μmol / L gentiopicroside was added for 1 h, then 10-6mol / L angiotensin II was added, and the medium was replaced after 24 h, and the treatment was performed for 48 h in total; -6 mol / L angiotensin II, and the medium was replaced after 24 h, and the treatment was performed for 48 h in total;
[0051] Lentinan group: 100 mg / L lentinan was added for 1 h, then 10-6mol / L angiotensin II was added, and the medium was replaced after 24 h, and the treatment was performed for 48 h in total; -6 mol / L angiotensin II, and the medium was replaced after 24 h, and the treatment was performed for 48 h in total;
[0052] Gentiopicroside and lentinan group: 20 μmol / L gentiopicroside + 100 mg / L lentinan was added for 1 h, then 10-6mol / L angiotensin II was added, and the medium was replaced after 24 h, and the treatment was performed for 48 h in total;
[0053] After the treatment, the cell culture solution was removed, the cells were washed with PBS, and the cells were fixed with glutaraldehyde for 10 min;
[0054] After washing with PBS for 3 times, the PBS was removed, 1 ml of X-gal staining solution was added to each well, and after incubation at 37°C for 12 h, 5 fields of view were randomly selected under a fluorescence microscope in each group, and the percentage of positive cells was calculated.
[0055] The experimental results of Example 1 are shown in Table 1:
[0056] Table 1 Influence of gentiopicroside on the activity of HUVEC cells
[0057] Note: * represents P<0.05
[0058] As can be seen from Table 1, when the concentration of gentiopicroside is 10 and 20 μmol / L, there is no obvious influence on the activity of HUVEC cells, and when the concentration of gentiopicroside is 40 μmol / L, it produces a certain degree of inhibition on the activity of HUVEC cells, therefore, the concentration of gentiopicroside is selected as 20 μmol / L to continue the subsequent experiments.
[0059] The experimental results of Example 2 are shown in Table 2:
[0060] Table 2 Influence of gentiopicroside on cell aging caused by angiotensin II
[0061] Note: *** represents P<0.001, represents P<0.001
[0062] As can be seen from Table 2, there is basically no cell aging in the control group, the proportion of positive cells in the angiotensin II group is significantly increased, and compared with the angiotensin II group, the proportion of positive cells in the gentiopicroside group is decreased to a certain extent. It shows that gentiopicroside can reduce the HUVEC cell aging caused by angiotensin II to a certain extent.
[0063] The results of Example 3 are shown in Figure 1 .
[0064] As can be seen from Figure 1 , the protein expression of Cav-1 in the angiotensin II group is increased, and compared with the angiotensin II group, the protein expression of Cav-1 in the gentiopicroside group is decreased to a certain extent, but is still higher than that in the control group, which is consistent with the results of Example 2, indicating that gentiopicroside can reduce the increase of Cav-1 protein expression caused by angiotensin II to a certain extent.
[0065] Results of Example 4 are shown in Table 3:
[0066] Note: *** means P<0.001, ** means P<0.01, *** means P<0.001
[0067] As can be seen from Table 3, compared with the combination of Gastrodin and Ganoderma lucidum polysaccharides alone, the combination of the two drugs can effectively produce a synergistic effect, thereby greatly reducing the vascular endothelial cell aging caused by angiotensin II.
Claims
1. A drug for delaying angiotensin II-induced vascular aging, characterized in that, The drug is a liquid drug, and the drug includes gentian bitter licorice, Ganoderma lucidum polysaccharide and a pharmaceutically acceptable carrier; The concentration of gentian bitter licorice in the drug is 20 μmol / L, and the concentration of Ganoderma lucidum polysaccharide in the drug is 100 mg / L.
2. A drug for reducing angiotensin II-induced vascular endothelial cell senescence, characterized in that, The drug is a liquid drug, and the drug includes gentian bitter licorice, Ganoderma lucidum polysaccharide and a pharmaceutically acceptable carrier; The concentration of gentian bitter licorice in the drug is 20 μmol / L, and the concentration of Ganoderma lucidum polysaccharide in the drug is 100 mg / L.
3. A culture medium for reducing angiotensin II-induced vascular endothelial cell senescence, characterized in that, The culture medium contains gentiopicrin and Ganoderma lucidum polysaccharide; The concentration of gentian bitter licorice in the culture medium is 20 μmol / L, and the concentration of Ganoderma lucidum polysaccharide in the culture medium is 100 mg / L.