Cordyceps militaris protein extract and antitumor application thereof
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A technology of protein extract and Cordyceps militaris, applied in the direction of antineoplastic drugs, peptide/protein components, applications, etc.
Active Publication Date: 2013-09-04
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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The current research progress has not yet found the report of extracting protein sub
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Embodiment 1
[0031] Embodiment 1, the preparation of Cordyceps militaris protein extract
[0032] 1. Preparation of fruiting bodies of Cordyceps militaris
[0033] Inoculate the slant strain of Cordyceps militaris (Anhui) CFCC89535 into the primary medium for activation, culture at a constant temperature of 25°C, and inoculate it into the secondary medium after the mycelium is full of the test tube, 25 ℃, 160r / min fermentation and culture for 10 days, inoculate the secondary liquid seed into the canned bottle containing the cultivation medium for 20 days under the condition of 20 ℃ and avoid light, after the mycelium grows to fill the whole bottle, give light and use Cold stimulation at 10°C promotes color change and the formation of sub-seats. The fruiting condition is to keep the humidity between 80-90% and the temperature at 18-25°C. Collect fruiting bodies and obtain Cordyceps militaris (Anhui) CFCC89535 subentity.
[0034] Wherein, the culture medium used in this experiment is as fo...
Embodiment 2
[0042] Example 2, Cordyceps militaris protein extract inhibits tumor cell proliferation experiment
[0043] 1.1 Tested cell lines
[0044] Human liver cancer HepG2 cells (purchased from ATCC, USA), human breast cancer MCF-7 cells (purchased from ATCC, USA) and human lung adenocarcinoma A-549 cells (purchased from ATCC, USA).
[0045] 1.2 Experimental method
[0046] 1.2.1 Cell lines and cell culture
[0047] Human liver cancer HepG2 cells, human breast cancer MCF-7 cells and human lung adenocarcinoma A-549 cells were activated and passaged according to conventional culture methods. Among them, the subculture and activation medium of human liver cancer HepG2 cells is DMEM-high glucose + 1% double antibody + 10% FBS (in DMEM-high glucose (Hyclone, SH30022.01B) a mixture of penicillin and streptomycin (penicillin 10000U / mL, streptomycin 10000μg / mL) and fetal bovine serum (FBS, Hyclone, SV30087.02), so that the final volume percentage of the former two is 1%, and the volume per...
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Abstract
The invention discloses a cordyceps militaris protein extract and an antitumor application thereof. The cordyceps militaris protein extract is prepared by a method comprising the following steps of: (1) crushing cordyceps militaris cystocarps, soaking with water, and collecting a water-soluble substance to obtain a cordyceps militaris water-soluble extract; and (2) performing ammonia sulfate precipitation of which the saturation degree is 80% on the cordyceps militaris water-soluble extract, collecting a deposit, and dialyzing the deposit by using deionized water to obtain the cordyceps militaris protein extract. The IC50 values, after the cordyceps militaris protein extract is used for treating human hepatic carcinoma HepG2 cells, human mammary cancer MCF-7 cells and human lung adenocarcinoma A-549 cells for 72 hours, respectively are 10 microgram/mL, 14 microgram/mL and 8 microgram/mL, and the inhibition rates are increased along with the expansion of time and increase of concentration; and the changes of apoptotic forms can be observed under a light microscope.
Description
technical field [0001] The invention relates to a protein extract of Cordyceps militaris and its anti-tumor application. Background technique [0002] Cordyceps militaris (Cordyceps militaris), which belongs to Ascomycota, Sclerotinia, Ergotomycetes, Ergotaceae, Cordyceps, is a precious medicinal fungus. The chemical composition and pharmacological effects of Cordyceps militaris are similar to those of Cordyceps sinensis, and it is often used as medicine instead of Cordyceps sinensis in clinical practice. Cordyceps militaris has the functions of sterilization, anti-inflammation, and immunity enhancement. It is not only edible, but also can be made into health care products and medicines. When artificially cultivating Cordyceps militaris, the yield rate often decreases. Wang Hexiang et al. focused on the methods of isolating wild Cordyceps militaris strains and screening high-yielding strains to improve the commercial efficiency of artificial cultivation. Lin Jinsheng and o...
Claims
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