Application of luteolin in preparation of ovarian senescence resistance medicines
A technology of luteolin and ovarian aging, which is applied in the direction of drug combinations, pharmaceutical formulas, and medical preparations containing active ingredients, etc., to achieve significant proliferation, expand the scope of application, and increase the secretion level
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Embodiment 1
[0016] [Example 1] Proliferation of luteolin on isolated ovarian granulosa cells (MTT method)
[0017] Female rats aged 22-27 days were killed by cervical dislocation after 48 hours of routine feeding. The bilateral ovaries of the rats were aseptically removed, and the ovaries were crushed to release granulosa cells. They were treated with 2 mL of DMEM-F12 containing penicillin and streptomycin. Dilute in culture medium and gently pipette to disperse into single cells. Pellet cells were collected by centrifugation. A certain concentration of cell suspension was inoculated in a 24-well plate, 80 μl per well, and 120 μl of culture medium was added to each well to adjust the cell density to 200 μl / well, and 6 parallel wells were set up for each group. Put it in 5% CO 2 Cultivate in the incubator for 24h. Take CO 2 The granule cell suspension was cultivated in the incubator for 24 hours, and each well was successively added with cadmium chloride of modeling concentration and d...
Embodiment 2
[0021] [Example 2] The effect of luteolin on the estrogen and progesterone secretion of isolated ovarian granulosa cells (radioimmunization method)
[0022] Female rats aged 22-27 days were killed by cervical dislocation after 48 hours of routine feeding. The bilateral ovaries of the rats were aseptically removed, and the ovaries were broken to release granulosa cells. They were treated with 2ml of DMEM-F12 containing penicillin and streptomycin. Dilute in culture medium and gently pipette to disperse into single cells. Pellet cells were collected by centrifugation. A certain concentration of cell suspension was inoculated in a 24-well plate, 80 μl per well, and 120 μl of culture medium was added to each well to adjust the cell density to 200 μl / well, and 6 parallel wells were set up for each group. Put it in 5% CO 2 Cultivate in the incubator for 24h. Take CO 2 The granule cell suspension was cultivated in the incubator for 24 hours, each well was successively added with ...
Embodiment 3
[0026] [Example 3] Effect of luteolin on the secretion of estrogen and progesterone in natural aging rats (radioimmunization method)
[0027] Adopt the pre-aging model of natural aging. Select 14-month-old female SD rats with a body weight of (270±30) g. After the animals are fed for one week, do vaginal cytology smears and observe 4 estrous cycles continuously. Vaginal cytology shows that the estrous cycle is prolonged and then continues to be in estrus , Repeated pseudo-pregnancy cell phase, as a successful model of female aging. Natural aging rats were randomly divided into model group, luteolin (10mg / kg, 30mg / kg, 50mg / kg) group and positive drug group, with 10 rats in each group, and another 11 4-month-old female rats were used as The normal control group was intragastrically administered once a day, and the normal control group and the model group were given equal volumes of normal saline. After 30 days of continuous administration, blood was collected from the orbital v...
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