Method of inducing apoptosis and compositions therefor
A composition and apoptosis technology, applied in the field of induction of apoptosis, can solve the problems of difficult digestion of cell body components, inability to absorb macromolecular proteins and polysaccharides, etc.
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Embodiment 1
[0062] 2L of distilled water and 300g of Kyowa Brazil mushrooms were mixed, heated to boiling, and refluxed for 2 hours. The above solution is filtered to obtain filtrate (hot water extract) and residue. Then 2L of distilled water and residue were mixed, heated to boiling, and refluxed for 2 hours to prepare hot water extract and residue. The above steps can be repeated once. The obtained filtrates were combined and freeze-dried to prepare a dry product A [153 g: extraction rate (recovery) 51%].
[0063] 500 mL of distilled water and 50 g of dry product A were mixed and dialyzed in a dialysis tube (Spectra / PoreMembrane 50×31, inner diameter 8 mm, length 30 cm, FE-0526-65). The mixture was dialyzed against 3 liters of distilled water for 12 hours. The dialysate was lyophilized to obtain a dry product C (27 g: extraction rate 53%). The remaining solution in the dialysis tubing was further dialyzed against hot water for 30 hours, and then dialyzed against distilled water twic...
Embodiment 2
[0067] A hot water extraction similar to that described in Example 1 was performed to obtain 6 L of combined filtrates (hot water extracted solution). Concentrate the filtrate to 1 L under reduced pressure, add 1 L of ethanol and mix to separate the polysaccharide. The mixture was centrifuged, and the precipitate and supernatant were separated. 3 L of ethanol was further mixed with the supernatant, centrifuged to obtain a precipitate, which was dissolved in distilled water and dialyzed. The dialysate was lyophilized to obtain a powder called ABMK-22.
Embodiment 3
[0069] The cell growth inhibitory activity (test 1), cell differentiation induction activity (test 2) and apoptosis induction activity (test 3) of agaricus hot water extract (ABMK-22) lyophilized powder were observed.
[0070] The biological activity test was carried out by the following method.
[0071] 1. Cell growth inhibitory activity (Test 1): In the analysis, the culture-drug sensitivity test (CD-DST method) embedded in collagen gel drops was used, and the cells were counted.
[0072] The CD-DST test was performed referring to the method of H. Kobayashi et al. (International Journal of Cancer 11: 449-455, 1997).
[0073] Briefly, the method was performed as follows: first, a cell (HL-60 cell line) suspension was mixed with collagen (such as type I collagen (Cellmatrix Type CD, Nitta Gelatin Inc.)), reconstitution buffer, medium (such as concentrated F-12 medium) mixed on ice, the test cells coated in collagen. The above mixed solution was placed in a multi-well culture...
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