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A kind of extraction method of gecko anticancer extract

An extraction method and extract technology, applied in the field of gecko anti-cancer extracts, can solve the problems that the tumor inhibitory effect is difficult to reach the tumor inhibitory level of fluorouracil and doxorubicin, and the extraction method is complicated, so as to achieve good anticancer effect and improve Immunity, the effect of simple preparation method

Inactive Publication Date: 2014-10-29
HENAN UNIV OF SCI & TECH
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  • Application Information

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Problems solved by technology

[0003] The above-mentioned large number of research results show that gecko extract has positive anti-tumor effect, and has a good application prospect in the treatment of solid tumors, but the current extraction method of gecko extract is relatively complicated, and gecko extract has a tumor-inhibiting effect on tumors. It is difficult to achieve the tumor inhibitory level of fluorouracil and doxorubicin

Method used

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  • A kind of extraction method of gecko anticancer extract
  • A kind of extraction method of gecko anticancer extract
  • A kind of extraction method of gecko anticancer extract

Examples

Experimental program
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Embodiment

[0017] 1. Drying and crushing of the gecko: After drying 1000g of the washed dry gecko without webs, crush it into a coarse powder with a pulverizer.

[0018] 2. Soaking, crushing and homogenization: Place in 4000mL Tris-HCl buffer solution (pH=7.4), soak overnight, ultrasonically break the cells, homogenize with a colloid mill and collect the homogenate.

[0019] 3. Centrifugation and alcohol extraction: Centrifuge at 7000g (11000rpm) for 20min to obtain 3800ml of supernatant, slowly add 4676ml of dehydrated ethanol (99.7%) to the supernatant solution to make the final concentration of ethanol 55%, let stand at 4°C for 3- After 4 hours, stir once every 30 minutes, then centrifuge at 7000 g (11000 rpm) for 20 minutes, and take the supernatant to obtain an aqueous solution of gecko ethanol extract.

[0020] 4. Concentration by rotary evaporation: Concentrate the aqueous alcohol extract solution to 200ml at 55°C and 0.085-0.1MPa.

[0021] 5. Salt dissolution and salting out: Sl...

experiment example

[0025] 1. Inhibitory effect of different concentrations of gecko anticancer extracts on the proliferation of human liver cancer cells HepG2

[0026] MTT method was used to detect the growth inhibition of gecko anticancer extracts on human hepatoma cell HepG2: the hepatoma cell HepG2 cultured in RPMI1640 medium was 1×10 per well. 4 / mL were inoculated in 96-well plates, and treated with different concentrations (0.625, 0.75, 1.5, 3mg / ml) of gecko anticancer extracts for 24, 48, and 72 hours, respectively. 4 hours before the end, add 20 μL of freshly prepared 5 mg / mL MTT solution to each well, continue to cultivate for 4 hours, discard the medium, add 150 μL of dimethyl sulfoxide (DMSO) to each well, and measure it with a microplate reader at a wavelength of 490 nm after it is completely dissolved. Absorbance A of each well, and calculate the inhibition rate according to the following formula. Inhibition rate (%)=[1-(A experimental group / A negative control)]×100%. The experime...

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Abstract

The invention discloses an anti-cancer extractive of wall gecko. A preparation method of the extractive comprises the following steps of: firstly, drying and smashing the wall gecko, soaking in a buffer solution with the pH of 7.4 for 12 hours, carrying out ethanol extraction by adding absolute ethyl alcohol, concentrating the extracting solution in an evaporation way, salting out by adding ammonium sulfate in the concentrated solution, obtaining white floccule precipitate in a dialyzing way, and drying the white floccule precipitate to obtain the anti-cancer extractive of the wall gecko. The anti-cancer extractive of the wall gecko disclosed by the invention is obtained by extracting the peptide molecules with the function of tumor resistance from the wall gecko by means of alcohol extraction, salt precipitation and dialysis, so that the extractive is very high in anti-cancer effect, thereby being capable of being taken as the active ingredients of high-efficiency and low-toxicity anti-cancer new drug. The test shows that the anti-cancer extractive of the wall gecko disclosed by the invention is very good in anti-cancer function, which reaches the anti-cancer level of the normal anti-cancer medicine such as fluorouracil and adriamycin, so that the immunity of the human body can be improved, and the extractive of the wall gecko contains the natural wall gecko polypeptide, so that the anti-cancer extractive has the advantages of being low in toxicity, and small in side effects.. Furthermore, the preparation method of the extractive is simple, thereby being easy to popularize and use.

Description

technical field [0001] The invention relates to a gecko anticancer extract. Background technique [0002] Gecko is a traditional Chinese medicinal material for treating tumors in my country. A large number of clinical reports have shown that gecko has a significant effect on treating malignant tumors. Dried gecko, fresh gecko, serum containing gecko, gecko compound preparation, gecko protease hydrolyzate, etc. all have good curative effects on tumors. Xie Shuang, Wang Xuemei, etc. (Xie Shuang, Wang Xuemei, Xie Dongze. Fresh Gecko Extract Inhibited C6 Glioma Cell Proliferation and Induced Cell Apoptosis. Tumor Prevention and Treatment Research, 2003, 30:458-461) used different concentrations of fresh gecko The extract (take the live gecko and freeze it to death, wash it with water, pulverize the homogenate, freeze and thaw it 10 times, get the supernatant by natural precipitation, centrifuge at 10000r / min at low temperature, centrifuge for 20min, take the supernatant) for C6...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/00A61K35/56A61K9/19A61P35/00A61K35/583
Inventor 王建刚宋莹崔朝初段冷昕李瑞芳
Owner HENAN UNIV OF SCI & TECH