Application of stigmasterol to preparation of medicine for improving cardiac hypertrophy
A technique of myocardial hypertrophy and stigmasterol, which is applied in the field of biomedicine, can solve the problems of poor therapeutic effect of myocardial hypertrophy and increased risk of sudden death of patients, and achieves the goal of inhibiting myocardial hypertrophy, inhibiting oxidative stress of myocardial cells, and improving myocardial hypertrophy. Effect
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Embodiment 1
[0044] Cytotoxicity detection of embodiment 1 stigmasterol
[0045] 1. Culture and passage of H9c2 cells
[0046] In an incubator maintained at constant temperature and humidity, culture was carried out with an appropriate proportion of Dulbecco's Modified Eagle's Medium (DMEM). When the cells were in good condition and the cell density reached 80-90%, subculture was carried out, the culture supernatant was discarded, washed twice with high-pressure phosphate buffered saline (PBS) with a pipette gun, and 1 mL of trypsin was absorbed for digestion. When most of the cells were observed to shrink under the microscope, 2 mL of medium was added to the ultra-clean bench to stop the digestion, the adherent cells were blown off with a pipette gun, and transferred to a 5 mL EP tube for centrifugation at 1500 rpm / min for 5 min. Discard the supernatant, add 2mL medium to resuspend, and carry out passage according to the ratio of 1:3.
[0047] 2. Cell Viability Detection
[0048] Cell ...
Embodiment 2
[0053] Embodiment 2 in vitro exploration experiment
[0054] 1. Immunofluorescence of cells
[0055] H9c2 cells were pretreated with 0.5 μg / mL (low), 2.5 μg / mL (medium) and 5 μg / mL (high) stigmasterol (STG) for 30 min, and then stimulated with ISO for 48 hours to observe the effect of stigmasterol on ISO-induced H9c2 cells. The effect of cardiomyocyte surface area, the specific steps are as follows:
[0056] (1) Take H9c2 cardiomyocytes in a good growth state, digest and resuspend them into a cell suspension, adjust the cell density to 6000 per well, plant them in a 48-well plate (the well plate is equipped with a small round glass slide with a radius of 4mm), About 12 hours after the cells were completely adhered to the wall, they were divided into blank, model, and administration groups (stigmasterol was added at least 30 minutes in advance in the administration group), and 10 μM ISO was added to each well of the model and administration groups for 48 hours;
[0057] (2) A...
Embodiment 3
[0082] Example 3 Stigmasterol inhibits the oxidative stress of ISO-induced H9c2 mast cells
[0083] A large number of studies have shown that the process of cardiac hypertrophy is often accompanied by an increase in oxidative stress. Therefore, the present invention adopts malondialdehyde MDA kit (article number A003-2) and superoxide dismutase SOD kit (article number A001-3) produced by Nanjing Jiancheng Company to detect the impact of ISO on the oxidative stress of H9c2 cells respectively. As well as the intervention effect of stigmasterol on the oxidative stress caused by ISO, the specific steps are carried out with reference to the instructions. The result is as image 3 As shown in A and 3B, compared with the CON group, the level of MDA in the cells of the ISO group was significantly increased, while the level of SOD was significantly reduced; compared with the ISO group, the stigmasterol pretreatment group, especially the high-dose treatment group (5μg / mL), can signif...
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