Semen trigonellae protein, separation and extraction method thereof and application thereof in preparation of bone marrow HSC removal adjuvant
An extraction method and the technology of fenugreek, which are applied in the field of natural functional products, can solve the problems of undefined functions of fenugreek plant protein macromolecules, and achieve the effects of easy industrial promotion and application, easy quality control, and wide sources
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Embodiment 1
[0030] The separation and extraction method of fenugreek protein comprises the following steps:
[0031] (1) Grind fenugreek, first soak it in petroleum ether to degrease, then add KH 2 PO 4 -NaOH buffer solution (pH value is 7; the same below) is stirred and extracted for 4-12 hours, centrifuged, and the supernatant is taken; the above-mentioned extraction operation is repeated several times, and the obtained supernatant is combined to be the extract;
[0032] (2) Slowly add ammonium sulfate to 40% saturation in the extract, centrifuge (2000r / min, centrifugal 15min; the same below), collect the supernatant; continue to add ammonium sulfate to 50% saturation, centrifuge, collect precipitation;
[0033] (3) Transfer the precipitate to a dialysis bag, dialyze in deionized water (change the water every 12 hours on average), until the dialysate is colorless, and finally freeze-dry the components in the dialysis bag to obtain fenugreek protein;
[0034] Give the fenugreek protei...
Embodiment 2
[0042] The separation and extraction method of fenugreek protein is the same as in Example 1, and the difference is step (2):
[0043] (2) Slowly add ammonium sulfate to 50% saturation in the extract, centrifuge, and collect the supernatant; continue to add ammonium sulfate to 60% saturation, centrifuge, and collect the precipitate;
[0044] Give the fenugreek protein precipitated by the ammonium sulfate saturation of 50%-60% prepared in this embodiment, the dose is 3.0g / kg.d, and continue for 28 days, and observe the effect of pretreatment on bone marrow transplantation of thalassemia iron overload model mice Influence of bone marrow depletion.
[0045] Experimental method is the same as embodiment 1.
[0046] As can be seen from Table 1, compared with the Bu / Cy group, giving the fenugreek protein of this embodiment at a dose of 3.0g / kg.d for 28 days increased the effect on G in the LSK cell population. 0 scavenging effect, manifested as G in the LSK cell population 0 The pr...
Embodiment 3
[0048] The separation and extraction method of fenugreek protein is the same as in Example 1, and the difference is step (2):
[0049] (2) Slowly add ammonium sulfate to 60% saturation in the extract, centrifuge, and collect the supernatant; continue to add ammonium sulfate to 70% saturation, centrifuge, and collect the precipitate;
[0050] Give the fenugreek protein precipitated by the ammonium sulfate saturation of 60%-70% prepared in this embodiment, the dose is 2.0g / kg.d, continue for 28 days, and observe the effect of pretreatment on bone marrow transplantation of thalassemia iron overload model mice Influence of bone marrow depletion.
[0051] Experimental method is the same as embodiment 1.
[0052] As can be seen from Table 1, compared with the Bu / Cy group, giving the fenugreek protein of the present embodiment at a dose of 2.0g / kg.d for 28 days increased the effect on G in the LSK cell population. 0 scavenging effect, manifested as G in the LSK cell population 0 T...
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