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Daylily antitumor active protein as well as preparation method and application thereof

A technology of anti-tumor activity and day lily, which is applied in the field of preparation and application of plant protein, can solve the problems of less anti-tumor active protein of day lily, insufficient systematic and in-depth chemical and pharmacological research of day lily, and achieve good anti-tumor cells active effect

Active Publication Date: 2021-03-26
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the chemical and pharmacological research on day lily at home and abroad is not systematic and in-depth.
And there are few domestic and foreign research reports on the anti-tumor active protein of day lily

Method used

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  • Daylily antitumor active protein as well as preparation method and application thereof
  • Daylily antitumor active protein as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The preparation of day lily anti-tumor active protein comprises the following steps:

[0022] Step 1: Take 200g of dried day lily, cut it into pieces with scissors, add 1L of phosphate buffer (1L of phosphate buffer contains 8g NaCl, 0.2g KCl, 3.63g NaCl 2 HPO4.12H 2 O, 0.24g KH 2 PO 4 , pH7.4), soaked and stirred overnight at 4°C, centrifuged at 11000rpm for 30min at 4°C, and took the supernatant, which was the crude day lily protein extract. The centrifuged filter residue can be repeatedly extracted 1-2 times;

[0023] Step 2: Add 5 times the volume of acetone pre-cooled at -20°C to the crude daylily protein extract in the first step, stir while adding, put it in a refrigerator at -20°C for 2 hours to precipitate, and centrifuge at 4°C at 11,000rpm After 30 minutes, the supernatant was discarded, the precipitate was collected, washed with acetone for 3 times, and the precipitate was air-dried to obtain day lily crude protein.

[0024] The third step: dissolve the...

Embodiment 2

[0030] The effect of day lily anti-tumor active protein on the survival of tumor cells

[0031] Liver cancer HepG2 and Bel-7402 cells in logarithmic growth phase, normal liver cell HL7702, colorectal cancer DLD1 cells, breast cancer MCF-7 cells, cervical cancer HeLa cells and ovarian adenocarcinoma SKOV3 cells were divided into 5×10 3 per well into a 96-well culture plate. 37°C with 5% CO 2 After incubating in the incubator for 24 hours, the day lily anti-tumor active protein of Example 1 was added to make the final concentrations respectively 2.5 μg / mL, 5 μg / mL, 20 μg / mL, 50 μg / mL, 100 μg / mL, and 150 μg / mL. Six replicate wells were set up for each concentration, and the culture medium containing the same volume of Tris-Hcl buffer was used as a control. After incubation for 24 hours, 20 μL of MTT solution (5 mg / mL) was added to each well, and incubation was continued for 4 hours before the culture was terminated. Carefully aspirate and discard the culture supernatant in the...

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Abstract

The invention relates to the field of preparation and application of vegetable protein, and particularly belongs to daylily antitumor active protein as well as a preparation method and application thereof. The specific preparation method comprises the following steps: cutting daylily into pieces, adding a protein extracting solution, performing stirring overnight at 4 DEG C, adding precooled acetone to precipitate protein at low temperature, and carrying out dissolution with the protein extracting solution after centrifugation to obtain crude daylily protein; and separating and purifying antitumor active components in the daylily crude protein by adopting ammonium sulfate fractional precipitation and cation exchange chromatography to obtain the daylily antitumor active protein. The proteincan obviously inhibit the survival of liver cancer HepG2 and Bel-7402 cells, colorectal cancer DLD1 cells, breast cancer MCF-7 cells, cervical cancer HeLa cells and ovarian adenocarcinoma SKOV3 cells, and can be applied to the preparation of antitumor drugs.

Description

technical field [0001] The invention relates to the field of preparation and application of plant protein, in particular to a day lily anti-tumor active protein and its preparation method and application. Background technique [0002] Day lily (Hemerocallis citrina Baroni), a perennial herb of the Liliaceae family, is also known as day lily, forget care grass, and day lily. It is a health-care vegetable with high nutritional value. Day lily can not only be tasted as a high-end dish, but more importantly, its unique nutrients can be made into a variety of supplements for brain, blood pressure, diuresis, nerves, and cancer prevention. Day lily contains carbohydrates, proteins, vitamins, inorganic salts and a variety of essential amino acids. Studies have found that day lily has a certain potential in anti-tumor. Day lily extracts such as 2-hydroxy chrysophanol and rhein can inhibit the proliferation of cancer cells. In addition, the colchicine contained in day lily has obvio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C07K1/14C07K1/18C07K1/30C07K1/34C07K1/36A61K38/16A61P35/00
CPCC07K14/415A61P35/00A61K38/00
Inventor 宋莉尤敏
Owner SHANXI UNIV
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