CN104887724A - Method for preparing extract of corchorus capsularis seeds and application of extract - Google Patents
Method for preparing extract of corchorus capsularis seeds and application of extract Download PDFInfo
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- CN104887724A CN104887724A CN201510230874.0A CN201510230874A CN104887724A CN 104887724 A CN104887724 A CN 104887724A CN 201510230874 A CN201510230874 A CN 201510230874A CN 104887724 A CN104887724 A CN 104887724A
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Abstract
The invention relates to a method for preparing the extract of corchorus capsularis seeds and the application of the extract. According to the method, corchorus capsularis seeds are crushed, and the petroleum ether 5 times the weight of corchorus capsularis seeds is added at the room temperature for extracting and degreasing. After the degreasing process, corchorus capsularis seeds are subjected to reflux extraction for 1-3 times in the ethanol aqueous solution 5-10 times the weight thereof, and the reflux extraction process lasts for 1-3 hours each time. The obtained extract is collected as the ethanol extract of corchorus capsularis seeds and is stored at 20 DEG C. The extract can be used for inhibiting the cell proliferation of human bladder cancer cells. Compared with the prior art, the extract is more efficient in growth proliferation effect for the human bladder cancer cell line T24 and has less influence on normal bladder epithelial cells. Therefore, the extract has an important significance on the treatment of bladder cancers, the comprehensive utilization of corchorus capsularis seed resources in China, and the increment of incomes for corchorus capsularis growers.
Description
Technical field
The invention belongs to medical art, especially relate to a kind of preparation method and application of corchorus capsularis seed extract.
Background technology
Bladder cancer (Bladder Cancer) is the modal malignant tumor of urinary system, and sickness rate occupies world's malignant tumor the 9th, occupies China's urinary system malignant tumor first.Although the onset bladder cancer of 70-80% is non-infiltration (also referred to as the shallow property of table), postoperative easy recurrence, its relapse rate up to 70%, and have an appointment 15% patient evolution for invasive bladder cancer.Mainly adopt clinically at present transurethral resection of bladder tumor (TUR-BT) afterwards the chemotherapeutics such as regular intravesical mitomycin (MMC) to prevent recurrence and the progress of non-infiltration bladder cancer, but have efficiency low (being only 30-50%) and the problem such as toxic and side effects is large.Therefore, in the urgent need to developing the anti-bladder cancer drug of new high-efficiency low-toxicity to improve the chemotherapy effect of bladder cancer and to reduce toxic and side effects.
The cancer therapy drugs such as the paclitaxel extracted from plant, camptothecine, vincristine, due to the drug of first choice that mechanism of action is unique, anticancer therapeutic has become many treatments of cancer significantly, account for the market share of anticarcinogen 30%.Therefore from plant resources, find new antitumor drug or lead compound, become one of focus of current treatment of cancer research field.From poisonous plants, find PTS be regarded as a shortcut.
Corchorus capsularis (Corchorus capsularis) belongs to Tiliaceae Corchorus phloem fiber crop, for annual herb plant, another name Urtica cannabina L., green fiber crops, network fiber crops, water network fiber crops, wild mestha, circle fruit Corchorus olitorius L., circle capsule Corchorus olitorius L., bitter Folium Cannabis, cattle mud thatch, three pearl grass, sky Folium Perillae and boon are first-class.Corchorus capsularis originates in Tropical Asian, and drought is extensively cultivated, and its phloem fiber has the features such as moisture pick-up properties is good, the moisture that scatters and disappears is fast, is mainly used in weave gunnysack, burlap etc.Corchorus capsularis seed is poisonous, has by as treatment diarrhoea dysentery and furuncle toxic swelling, has clearing away summer-heat, the effect of detumescence of drawing out pus by applying a plaster to the affected part among the people.
By literature search, there is not yet corchorus capsularis seed extract so far for suppressing the report of proliferation of human bladder cancer cells and relevant disease aspect thereof.
Summary of the invention
Object of the present invention is exactly provide a kind of better efficacy to overcome defect that above-mentioned prior art exists and normal Urothelial Cell affected to preparation method and the application of little corchorus capsularis seed extract.
Object of the present invention can be achieved through the following technical solutions:
The preparation method of corchorus capsularis seed extract, adopts following steps:
(1) corchorus capsularis seed is pulverized, add the petroleum ether lixiviate defat at ambient temperature of its 5 times of weight;
(2) after defat completes, utilize the ethanol water of 5-10 times of weight to carry out reflux, extract, 1 ~ 3 time to corchorus capsularis seed, each 1 ~ 3h, collect the extractum obtained, be the ethanol extraction of corchorus capsularis seed, in-20 DEG C of preservations.
Corchorus capsularis seed after pulverizing is lixiviate defat 48h in petroleum ether.
Utilize the corchorus capsularis seed 2h of ethanol water at room temperature after airtight soak degreasing, then heating in water bath refluxes to reflux temperature, obtains extractum after filtration with Rotary Evaporators is concentrated.
The volume fraction of described ethanol water is 60-90%.
Described corchorus capsularis seed is wild or cultivation corchorus capsularis seed.
Prepare corchorus capsularis seed extract and can be applied to the suppression that human bladder cancer cell is bred, adopt following steps:
(1) cell of exponential phase is in: be inoculated in 96 orifice plates by human bladder cancer cell T24 with 10000, every hole cell concentration;
(2) spend the night adherent after, with 0.1,0.5,1,5 and the corchorus capsularis seed ethanol extraction administration of 10ug/ml concentration, and with 5,10,50 and 100ug/ml mitomycin for positive control, 6 repeating holes are established in each process, repeat experiment 3 times;
(3) after administration 24h, adopt mtt assay to detect human bladder cancer T24 cell proliferation rate of growth, and calculate IC50 value, and compare with the result of mitomycin, obtain the suppression result that corchorus capsularis seed extract is bred human bladder cancer cell T24.
Described human bladder cancer cell T24 buys from Chinese Academy of Sciences's cell bank.
Compared with prior art, the present invention is found by external MTT active anticancer evaluation, and corchorus capsularis seed extract is more efficient than clinical first-line drug mitomycin to the growth inhibited effect of human bladder cancer T24 cell, and little on normal Urothelial Cell impact.Therefore corchorus capsularis seed extract for the preparation of anti-bladder cancer drug, can have good development prospect.
The purposes of corchorus capsularis seed extract in preparation anti-bladder cancer drug belongs to first public, and corchorus capsularis seed ethanol extraction is more efficient than current clinical first-line drug mitomycin to the growth inhibition ratio of transitional cell bladder carcinoma cell line, and little on normal Urothelial Cell impact.Therefore corchorus capsularis seed ethanol extraction can for the preparation of the anti-bladder cancer drug of high-efficiency low-toxicity, there is good development prospect, to the comprehensive utilization of bladder cancer treatment, China's Corchorus olitorius L. germ plasm resource and to increase numb agriculture income all significant, in preparation process, petroleum ether lixiviate defat is adopted to make growth inhibition ratio have further raising.
Accompanying drawing explanation
Fig. 1 is the IC50 value after corchorus capsularis seed extract and mitomycin effect T24 cell 24h.
Fig. 2 is the suppression situation to T24 Growth of Cells after 5ug/ml corchorus capsularis seed extract and mitomycin effect 24h.
Detailed description of the invention
Below in conjunction with the drawings and specific embodiments, the present invention is described in detail.
Embodiment 1
Prepare the ethanol extraction of corchorus capsularis seed
Get 100g corchorus capsularis seed, pulverize, add the petroleum ether room temperature lixiviate defat 48h of 500ml, then use the ethanol water of 1000ml 70%, under room temperature after airtight immersion 2h, heating in water bath is to backflow, and each reflux, extract, 3h, is total to reflux, extract, 2 times.Collect the ethanol extraction of 2 times, after filtration, concentrate to obtain extractum with Rotary Evaporators.Collect extractum (being the ethanol extraction of corchorus capsularis seed) ,-20 DEG C save backup.
Embodiment 2
The inhibitory action experiment that corchorus capsularis seed ethanol extraction is bred human bladder cancer cell
Be in the cell of exponential phase: human bladder cancer cell T24 (buying from Chinese Academy of Sciences's cell bank) is inoculated in 96 orifice plates with 10000, every hole cell concentration.
Spend the night adherent after, with 0.1,0.5,1,5 and the corchorus capsularis seed ethanol extraction administration of 10ug/ml concentration, and with 5,10,50 and 100ug/ml mitomycin for positive control.6 repeating holes are established in each process, repeat experiment 3 times.
After administration 24h, adopt mtt assay to detect human bladder cancer T24 cell proliferation rate of growth, and calculate IC50 value.
As shown in Figure 1, after effect T24 cell 24h, IC50 (the growth half suppresses) dosage of corchorus capsularis seed extract is 2.44ug/ml, be 16.6% of mitomycin 14.70ug/ml, reach pole significant difference, the more efficient to the current clinical first-line drug mitomycin of the proliferate rejection ratio of transitional cell bladder carcinoma cell line T24 of corchorus capsularis seed extract is described, there is good development prospect.As shown in Figure 2, after effect 24h, 5ug/ml corchorus capsularis seed extract is 63.36% to the growth inhibition ratio of T24 cell, is 2.08 times of mitomycin 30.39%, reaches pole significant difference.These results suggest that the current clinical first-line drug mitomycin of the anti-bladder cancer specific activity of corchorus capsularis seed extract is more efficient, for the preparation of anti-bladder cancer drug, good development prospect can be had.
Embodiment 3
The preparation method of corchorus capsularis seed extract, adopts following steps:
(1) wild corchorus capsularis seed is pulverized, add the petroleum ether lixiviate defat 48h at ambient temperature of its 5 times of weight;
(2) after defat completes, utilize 5 times of weight, volume fraction is the corchorus capsularis seed 2h of 90% ethanol water at room temperature after airtight soak degreasing, then heating in water bath to reflux temperature carries out reflux, extract, 1 time, extraction time is 3h, collect the extractum obtained, be the ethanol extraction of corchorus capsularis seed, in-20 DEG C of preservations.
Embodiment 4
The preparation method of corchorus capsularis seed extract, adopts following steps:
(1) corchorus capsularis seed will be cultivated pulverize, add the petroleum ether lixiviate defat 48h at ambient temperature of its 5 times of weight;
(2) after defat completes, utilize 10 times of weight, volume fraction is the corchorus capsularis seed 2h of 60% ethanol water at room temperature after airtight soak degreasing, then heating in water bath to reflux temperature carries out reflux, extract, 1 time, extraction time is 3h, collect the extractum obtained, be the ethanol extraction of corchorus capsularis seed, in-20 DEG C of preservations.
Finally be necessary described herein: above embodiment is only for being described in more detail technical scheme of the present invention; can not be interpreted as limiting the scope of the invention, some nonessential improvement that those skilled in the art's foregoing according to the present invention is made and adjustment all belong to protection scope of the present invention.
Claims (8)
1. the preparation method of corchorus capsularis seed extract, is characterized in that, adopts following steps:
(1) corchorus capsularis seed is pulverized, add the petroleum ether lixiviate defat at ambient temperature of its 5 times of weight;
(2) after defat completes, utilize the ethanol water of 5-10 times of weight to carry out reflux, extract, 1 ~ 3 time to corchorus capsularis seed, each 1 ~ 3h, collect the extractum obtained, be the ethanol extraction of corchorus capsularis seed, in-20 DEG C of preservations.
2. the preparation method of corchorus capsularis seed extract according to claim 1, is characterized in that, the corchorus capsularis seed after pulverizing is lixiviate defat 48h in petroleum ether.
3. the preparation method of corchorus capsularis seed extract according to claim 1, it is characterized in that, utilize the corchorus capsularis seed 2h of ethanol water at room temperature after airtight soak degreasing, then heating in water bath refluxes to reflux temperature, obtains extractum after filtration with Rotary Evaporators is concentrated.
4. the preparation method of the corchorus capsularis seed extract according to claim 1 or 3, is characterized in that, the volume fraction of described ethanol water is 60-90%.
5. the preparation method of corchorus capsularis seed extract according to claim 1, is characterized in that, described corchorus capsularis seed is wild or cultivation corchorus capsularis seed.
6. preparation method as claimed in claim 1 prepares the application of corchorus capsularis seed extract, it is characterized in that, this extract is applied to the suppression to human bladder cancer cell's propagation.
7. the application of corchorus capsularis seed extract according to claim 6, is characterized in that, adopts following steps:
(1) cell of exponential phase is in: be inoculated in 96 orifice plates by human bladder cancer cell T24 with 10000, every hole cell concentration;
(2) spend the night adherent after, with 0.1,0.5,1,5 and the corchorus capsularis seed ethanol extraction administration of 10ug/ml concentration, and with 5,10,50 and 100ug/ml mitomycin for positive control, 6 repeating holes are established in each process, repeat experiment 3 times;
(3) after administration 24h, adopt mtt assay to detect human bladder cancer T24 cell proliferation rate of growth, and calculate IC50 value, and compare with the result of mitomycin, obtain the suppression result that corchorus capsularis seed extract is bred human bladder cancer cell T24.
8. the application of corchorus capsularis seed extract according to claim 7, is characterized in that, described human bladder cancer cell T24 buys from Chinese Academy of Sciences's cell bank.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104857038A (en) * | 2015-05-07 | 2015-08-26 | 同济大学 | Preparation method of corchorus olitorius seed extract and application |
| CN105498708A (en) * | 2015-12-16 | 2016-04-20 | 湖南广播电视大学 | Supercritical extraction method for jute polysaccharide and application of jute polysaccharide |
| CN105536712A (en) * | 2015-12-16 | 2016-05-04 | 湖南广播电视大学 | Ephedran prepared through steam explosion method and use thereof |
| CN104887724B (en) * | 2015-05-07 | 2018-12-18 | 同济大学 | The preparation method and application of corchorus capsularis seed extract |
| CN114847079A (en) * | 2022-05-17 | 2022-08-05 | 福建省农业科学院亚热带农业研究所（福建省农业科学院蔗麻研究中心） | Application of fructus cannabis sativa seeds in edible fungus cultivation |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104887724B (en) * | 2015-05-07 | 2018-12-18 | 同济大学 | The preparation method and application of corchorus capsularis seed extract |
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| CN102416019A (en) * | 2011-11-18 | 2012-04-18 | 桂林医学院 | Application of cedilanid in preparation of anti-liver cancer and anti-bladder cancer medicament |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104857038A (en) * | 2015-05-07 | 2015-08-26 | 同济大学 | Preparation method of corchorus olitorius seed extract and application |
| CN104857038B (en) * | 2015-05-07 | 2018-10-26 | 同济大学 | The preparation method and application of corchorus olitorius seed extract |
| CN104887724B (en) * | 2015-05-07 | 2018-12-18 | 同济大学 | The preparation method and application of corchorus capsularis seed extract |
| CN105498708A (en) * | 2015-12-16 | 2016-04-20 | 湖南广播电视大学 | Supercritical extraction method for jute polysaccharide and application of jute polysaccharide |
| CN105536712A (en) * | 2015-12-16 | 2016-05-04 | 湖南广播电视大学 | Ephedran prepared through steam explosion method and use thereof |
| CN105536712B (en) * | 2015-12-16 | 2018-03-06 | 湖南广播电视大学 | Jute polysaccharide and its application made from steam explosion method |
| CN105498708B (en) * | 2015-12-16 | 2018-03-06 | 湖南广播电视大学 | The means of supercritical extraction of jute polysaccharide and its application |
| CN114847079A (en) * | 2022-05-17 | 2022-08-05 | 福建省农业科学院亚热带农业研究所（福建省农业科学院蔗麻研究中心） | Application of fructus cannabis sativa seeds in edible fungus cultivation |
| CN114847079B (en) * | 2022-05-17 | 2023-12-12 | 福建省农业科学院亚热带农业研究所（福建省农业科学院蔗麻研究中心） | Application of jute seeds with round fruits in edible fungus cultivation |
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