Application of fagomine in preparation of anti-ovarian senescence drug
A technology of ovarian aging and buckwheat alkaloid, which is applied in the application field of buckwheat alkaloid in the preparation of anti-ovarian aging drugs, and achieves the effects of improving secretion level, significantly proliferating and increasing the secretion of progesterone.
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Embodiment 1
[0017] [Example 1] Proliferation effect of buckwheat base on isolated ovarian granulosa cells (MTT method)
[0018] 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. Dilute in F12 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 h, and each well was successively added with the concentration of cadmium chloride and samples of different concentrations of buckwheat alkali (2.5 mg·mL -1 , 0.5mg·mL ...
Embodiment 2
[0020] [Example 2] The effect of buckwheat on the secretion of estrogen and progesterone in isolated ovarian granulosa cells (radioimmunization method)
[0021] Female rats aged 22-27 days were sacrificed 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 2ml of DMEM containing penicillin and streptomycin- Dilute in F12 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 hole was added with cadmium chlo...
Embodiment 3
[0023] [Example 3] Effect of Buckwheat Alkaline on Estrogen and Progesterone Secretion in Natural Aging Rats (Radioimmunoassay)
[0024]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. The natural aging rats were randomly divided into model group, buckwheatine (10mg / kg, 30mg / kg, 50mg / kg) group and positive drug group, with 10 rats in each group, and another 11 female rats aged 4 months were taken as normal The 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 orbi...
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