Application of hickory nut endocarp extract in the preparation of anti-ovarian cancer products
A hickory nut and endocarp technology, applied in the application field of hickory nut endocarp extract in the preparation of anti-ovarian cancer products, can solve the problem of anti-cancer and ovarian cancer of hickory nut endocarp that has not been seen, and achieve good anticancer activity , the effect of high selective toxicity
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Embodiment 1
[0041] Embodiment 1: the preparation of pecan endocarp extract
[0042] (1) Selection and processing of raw materials
[0043] Select Linan local hickory nuts that are fully mature, smooth, crack-free, and odor-free. Soak the hickory nuts in warm water at about 40°C for 30 minutes, so that the seeds and seed coats can be peeled off more conveniently. Remove the shell to separate the pulp from the shell (exocarp and mesocarp). After obtaining the hickory nut kernel, the seeds and the seed coat are manually peeled off. Mesh sieve to obtain hickory nut endocarp powder, seal it with a sealed bag, and store it at -80°C for later use.
[0044] (2) Preparation of Hickory nut endocarp extract
[0045] The hickory nut endocarp powder was dissolved with 80v / v% methanol aqueous solution according to the ratio of material to liquid 1g:10ml, and extracted with the aid of ultrasonic waves, the extraction power was 240W, the extraction temperature was 25°C, and the extraction time was 2h. ...
Embodiment 2
[0046] Example 2: Inhibitory Effect of Hickory Nut Endocarp Extract on Ovarian Cancer Cell Proliferation
[0047] Take 100uL of human ovarian cancer cell OVCAR-3 and normal ovarian cell IOSE 364, inoculate 10,000 cells / well in a 96-well plate (medium is RPMI-1640+10v / v% FBS), and incubate at 37°C, 5%CO 2Incubate for 16 h to attach the cells, remove the medium, and wash once with PBS buffer (0.01M, pH 7.4). The culture medium (the culture medium is RPMI-1640+10v / v% FBS) containing different concentrations of pecan endocarp extract was treated for 24h. Then remove the medium, wash twice with PBS buffer (0.01M, pH7.4), add 100 μL of freshly prepared AQueous One detection solution, incubate at 37°C for 1 h, and use a microplate reader to perform cell viability analysis at 490 nm.
[0048] The experiment used the AQueous One method to determine the inhibitory effect of hickory nut endocarp extract on human ovarian cancer cell OVCAR-3 and ovarian normal cell IOSE 364. Such as fi...
Embodiment 3
[0049] Example 3: Pecan endocarp extract induces apoptosis of ovarian cancer cells through p53-dependent endogenous pathway
[0050] Alexa Flour 488 Annexin (AnnexinV) Apoptosis Kit (Invitrogen) was used to detect the effect of hickory nut endocarp extract on apoptosis of ovarian cancer cells. OVCAR-3 and IOSE 364 cells were treated with 10 6 The concentration per dish was inoculated in a 60mm culture dish (the medium was RPMI-1640+10v / v% FBS), at 37°C, 5% CO 2 Incubate for 16 h, remove the medium. Then add the culture medium (the culture medium is RPMI-1640+10v / v%FBS) containing different concentrations of hickory nut endocarp extract at 37 ℃, 5% CO 2 After cultured for 24 hours, the cells were collected, stained by Annexin V / PI double staining method, and analyzed by flow cytometry after staining. Such as Figure 2-Figure 4 As shown, the increase in apoptosis of OVCAR-3 cells after hickory endocarp extract treatment was dose-dependent. When treated with 20μg / mL pecan en...
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