CN108272780A - It is a kind of that there is the composition for inhibiting hepatoma cell proliferation effect and its application - Google Patents
It is a kind of that there is the composition for inhibiting hepatoma cell proliferation effect and its application Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/05—Phenols
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
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Abstract
There is the composition for inhibiting hepatoma cell proliferation effect and its application the invention discloses a kind of, belong to antitumor drug technical field.The composition includes the resveratrol using a concentration of 12.5 μm of ol/L, 200 μm of ol/L and the curcumin using 200 μm of ol/L of a concentration of 12.5 μm of ol/L.The present invention has found the composition of the resveratrol curcumin obtained under certain concentration ratio to human liver cancer HepG by the rational proportion to resveratrol and curcumin2The inhibiting rate of cell Proliferation has synergistic effect.Under conditions of using a concentration of 12.5 μm of ol/L, the inhibiting rate that resveratrol is used alone is 6.51%, and it is 9.86% that curcumin, which is used alone,.However, used at the same time in the two, inhibiting rate can then reach 48.85%, be 2.99 times of the sum of inhibiting rate of above two component 16.34%.It follows that resveratrol has extremely significant synergistic effect with curcumin on the inhibiting rate being proliferated to human hepatoma HepG2 cell, the technique effect is far beyond the range that those skilled in the art are expected.
Description
Technical field
There is the composition for inhibiting hepatoma cell proliferation effect and its application the present invention relates to a kind of, belong to antitumor drug
Technical field.
Background technology
Liver cancer is clinically common one of alimentary system malignant tumour, and in China, incidence accounts for malignant tumour third position,
Case fatality rate accounts for second.Chemotherapy is still main treatment means at present, but liver cancer cells are to the refractoriness of chemotherapeutics, and
The drug resistance generated in the treatment causes liver cancer general curative effect bad.
More and more studies have shown that resveratrol has antitumaous effect, kinds cancer can be prevented and treated.White black false hellebore
Alcohol may act on the three phases of the starting of tumour, promotion, development, and can obviously inhibit the different cancerous cell lines of in vitro culture
Proliferation, such as lymphoid cell cancer, Huppert's disease, breast cancer, gastric cancer, colon cancer, cancer of pancreas, oophoroma and cervix cancer
Deng.
Curcumin is a kind of phenol sulfonate extracted from the rhizome of the woody plants such as turmeric, Radix Curcumae, curcuma zedoary and calamus,
With extensive pharmacological action, such as anti-inflammatory, anti-oxidant, removing free radical, resisting HIV, protection liver and kidney
The effects that dirty, anti-fiber and cancer-resisting, has the characteristics that safety, without notable adverse reaction.
In recent years, resveratrol or curcumin are combined with other chemotherapeutics, are enhanced antitumor action and are reduced chemotherapy
The effect of adverse drug reaction, has been to be concerned by more and more people.But the two drug combination treating cancer rarely has research.For example, existing
There are patent CN 105076486A to disclose a kind of reducing blood lipid peanut oil rich in curcumin and resveratrol.Although in the patent
Contain two kinds of ingredients of curcumin and resveratrol in product, but is only according to existing report for the use of both the above ingredient
Property added in the product to realize the diversification of product efficacy, the antitumous effect of product is not verified.Meanwhile
Since the product is the complex system containing Multiple components such as peanut, ox bone marrow, wheat embryos, can curcumin and resveratrol
Its effect is played completely not verify finally.
Invention content
To improve the inhibiting effect effect of resveratrol and curcumin to hepatoma cell proliferation, give full play to resveratrol and
Drug effect of the curcumin in treating liver cancer, the present invention provides a kind of composition for having and inhibiting hepatoma cell proliferation effect, institutes
The technical solution taken is as follows：
A kind of to have the composition for inhibiting hepatoma cell proliferation effect, the composition is including using a concentration of 12.5 μm of ol/
The curcumin of the resveratrol and a concentration of 12.5 μm of ol/L-200 μm of ol/L of use of L-200 μm of ol/L.
Preferably, a concentration of 12.5 μm of ol/L-50 μm of ol/L of the use of the resveratrol；The use of the curcumin is dense
Degree is 12.5 μm of ol/L-100 μm of ol/L.
It is highly preferred that the ratio between the concentration of the resveratrol and curcumin is：Resveratrol：Curcumin=1；(1-8).
It is highly preferred that the ratio between the concentration of the resveratrol and curcumin is：Resveratrol：Curcumin=1；(1-2.5).
It is highly preferred that a concentration of 12.5 μm of ol/L~30 μm ol/L of the use of the resveratrol.
The composition of any description above can be applied in preparing treatment medicines resistant to liver cancer.
Compared with the existing technology, the advantageous effect that the present invention obtains is：
The present invention has found the white lamb's-quarters obtained under certain concentration ratio by the rational proportion to resveratrol and curcumin
The composition of reed alcohol-curcumin is to human liver cancer HepG2The inhibiting rate of cell Proliferation has synergistic effect.Using a concentration of 12.5
Under conditions of μm ol/L, the inhibiting rate that resveratrol is used alone is 6.51%, and it is 9.86% that curcumin, which is used alone,.So
And it is used at the same time in the two, inhibiting rate can then reach 48.85%, be the sum of the inhibiting rate of above two component
2.99 times of 16.34%.It follows that resveratrol has with curcumin on the inhibiting rate being proliferated to human hepatoma HepG2 cell
There is extremely significant synergistic effect, the technique effect is far beyond the range that those skilled in the art are expected.
Description of the drawings
Fig. 1 is growth inhibition ratio (x ± SD, n=6) of the resveratrol of various concentration to hepatoma Hep G 2 cells for 24 hours.Its
In, administration group compared with blank control group, * P<0.05；#P<0.01.
Fig. 2 is that mtt assay detects growth inhibition ratio (x ± SD, n after various concentration resveratrol acts on HepG2 cells for 24 hours
=6).Wherein, administration group compared with blank control group, * P<0.05；#P<0.01.
Fig. 3 is that mtt assay detects growth inhibition after various concentration resveratrol joint curcumin acts on HepG2 cells for 24 hours
Rate (x ± SD, n=6).Wherein, administration group compared with blank control group, #P<0.01.
Specific implementation mode
Following embodiment material therefor, reagent, device, equipment, method and control process are this without specified otherwise
Field common material, reagent, device, equipment, method and control process, those skilled in the art can be obtained by commercial channel
, or conventional arrangement is carried out according to specific needs, without involving an inventive effort.
Human hepatoma cell strain HepG2Purchased from Chinese Academy of Sciences's Shanghai biochemistry and Institute of Cell Biology；Resveratrol, ginger
Flavine is purchased from Chinese drug and food identification research institute；MEM culture mediums, trypsase, fetal calf serum are purchased from Gibco companies of the U.S..
The dissolving of resveratrol and curcumin uses dimethyl sulfoxide (DMSO), is diluted using culture solution.
With reference to specific embodiment, the present invention will be further described, but the present invention should not be limited by the examples.
Embodiment 1
1.HepG2 cell strain cultures
HepG2 cell lines cryopreservation tube is taken out into cryopreservation tube, is immersed in 37 DEG C of water-baths, constantly shakes and it is enabled to melt as early as possible
Change, notices that nozzle have upstream face hereinafter, in order to avoid polluting；It waits for that cell melts, cryopreservation tube, 75% second is taken out from water-bath
Alcohol wiping freezes pipe surface, is put into superclean bench, cell suspension is transferred in 15mL centrifuge tubes, and 6-8mL is added and contains
The MEM fresh cultures of 10% fetal calf serum, 1500rpm centrifuge 2min；It discards supernatant, cell is resuspended with fresh culture, is turned
Static gas wave refrigerator in culture bottle is moved to, the temperature of cell incubator is 37 DEG C, CO2A concentration of 5%, humidity 100% is replaced every other day
Fresh culture.
2.MTT methods measure inhibitory rate of cell growth
By HepG2Concentration of cell suspension is adjusted to 5 × 106A/mL is inoculated in 96 orifice plates per 200 μ L of hole, is separately added into not
With the resveratrol (12.5,25,50,100,200 μm of ol/L) or curcumin (12.5,25,50,100,200 μm of ol/ of concentration
L) or 200 μ of (12.5 μm of ol/L resveratrols, curcumin 12.5,25,50,100 μm of ol/L) MEM culture solutions is added in two medicines joint
L function cells, blank control group add 200 μ L complete mediums, are affected to measurement result because curcumin color is relatively deep, separately set
Liquid control group (only adds culture medium, various concentration liquid is acellular).Every group sets 6 parallel holes, continues culture and MTT is added for 24 hours
(5mg/mL) 20 μ L are placed and are abandoned supernatant in incubator after 4h, and 200 μ L DMSO are added, and 15min are shaken, with each hole of enzyme linked immunosorbent detection
Absorbance value (OD values) at 490nm.Take the mean of 6 hole OD values by formula：Inhibiting rate (%)=[ABlank control(AExperiment
ALiquid compares)/(AControl)] × 100%.
3. statistical procedures
Data withIt indicates, using SPSS19.0 softwares, more each group difference of one-way analysis of variance, P<0.05
Or P<0.01 indicates that difference has statistical significance.
4. result
4.1 mechanism of resveratrol inhibiting HepG2The inhibiting effect of cell growth
Such as table 1, shown in Fig. 1, various concentration resveratrol acts on human liver cancer HepG2Cell for 24 hours after, as administration concentration increases
Cell growth inhibition is added to enhance, and a concentration of 25,50,100,200 μm of ol/L resveratrols administration groups and blank control groups
Compared to there were significant differences (P<0.05).
1 mechanism of resveratrol inhibiting HepG of table2Cell growth inhibiting effect (N=6)
Table is noted：Compared with blank control group is administered in (1) 12.5 μm of ol/L, 25 μm of ol/L resveratrols,*P<0.05。
(2) 50,100,200 μm of ol/L resveratrol administration group compared with blank control groups,#P<0.01。
4.2 curcumins inhibit HepG2The inhibiting effect of cell growth
Such as table 2, shown in Fig. 2, after various concentration curcumin acts on human hepatoma HepG2 cell for 24 hours, as administration concentration increases
Cell growth inhibition enhances, and each administration group equal there were significant differences (P with blank control group compared with<0.05).
2 curcumin of table inhibits HepG2Cell growth inhibiting effect (N=6)
Table is noted：Compared with blank control group is administered in (1) 12.5 μm of ol/L resveratrol,*P<0.05。
(2) 25,50,100,200 μm of ol/L resveratrol administration group compared with blank control groups,#P<0.01。
4.3 resveratrols combine the inhibiting effect that curcumin inhibits HepG2 cell growths
Such as table 3, shown in Fig. 3, resveratrol combines curcumin administration, and 12.5 μm of ol/L of fixed resveratrol concentration are added
Various concentration curcumin (12.5,25,50,100 μm of ol/L), after acting on human hepatoma HepG2 cell for 24 hours, as administration concentration increases
Cell growth inhibition is added to enhance, and each administration group there were significant differences (P compared with blank control group<0.05).
3 resveratrol of table combines curcumin and inhibits HepG2Cell growth inhibiting effect (N=6)
Table is noted：Resveratrol is 1 according to concentration ratio with curcumin:1,1:2,1:4,1:8 administrations are compared with blank control group
Compared with,#P<0.01。
By testing above, it is found that resveratrol or curcumin or two medicines combine the liver cancer HepG in vitro culture2Cell
Inhibited proliferation.After individually giving resveratrol for 24 hours, MTT the results show that resveratrol medicament it is a concentration of 25,50,100,
When 200 μm of ol/L, compares cell growth inhibition with blank control group and significantly increase, and concentration is bigger, inhibiting rate is higher.Individually give
After giving curcumin for 24 hours, MTT is the results show that various concentration curcumin group (12.5,25,50,100,200 μm of ol/L), with blank pair
Cell growth inhibition is compared according to group significantly to increase, and 12.5,25,50,100 μm of ol/L groups, increased with curcumin drug concentration, suppression
Rate processed increases.Based on the above experiment basis, 12.5 μm of ol/L of resveratrol concentration are then prepared, combine various concentration curcumin
(12.5,25,50,100 μm of ol/L) acts on HepG2Cell, it is 1 to make resveratrol and curcumin drug concentration ratio:1,1:2,
1:4,1:8.MTT is the results show that two medicine synergy, and compared with blank control group, inhibitory rate of cell growth has notable raising,
Also, when two medicine administering drug combinations concentration are respectively 12.5 μm of ol/L, inhibiting rate 48.85% is administered alone far above the two
12.5 μm of ol/L groups (6.51%, 9.86%), 25 μm of ol/L groups (17.97%, 19.07%) and 50 μm of ol/L groups (39.90%,
35.56%).Wherein, under 12.5 μm of ol/L concentration, the technique effect of resveratrol and curcumin synergy is far longer than two
The sum of the technique effect of person's distribution effect, resveratrol and curcumin are on the proliferation inhibiting effect to human hepatoma HepG2 cell
With synergistic effect.The result shows that resveratrol is administered alone with administering drug combinations with curcumin to HepG2Cell Proliferation has suppression
It makes and uses, administering drug combinations concentration is far below being administered alone concentration and inhibiting effect is stronger.Above-mentioned technique effect is far beyond this
The intended scope of field technology personnel.
Embodiment 2
To further increase the function and effect of resveratrol and curcumin, the present embodiment is dense to the use of the two synergy
Degree has carried out further preferably.It is 1 in both fixed concentration ratio:In the case of 1, changes the use concentration of the two, exist respectively
12.5 μm of ol/L, 15 μm of ol/L, 17.5 μm of ol/L, 20 μm of ol/L, 22.5 μm of ol/L, 25 μm of ol/L and 30 μm of ol/L concentration under
It measures every group to be determined the proliferation inhibition rate of human hepatoma HepG2 cell, the results are shown in Table 4.
The joint component of 4 same ratio various concentration of table is to HepG2Cell growth inhibiting effect (N=6)
As shown in Table 4, in resveratrol and curcumin with 1：In the case that 1 fixed proportion is used in combination, with
Using being continuously increased for concentration, the proliferation inhibition rate of human hepatoma HepG2 cell is continuously improved in drug, when concentration increases to 20 μ
When mol/L or more, the effect of synergy is than in embodiment 11：The inhibition of 8 ratio is slightly higher, but when concentration increases
To 22.5 μm of ol/L or more, the improvement effect of resveratrol and curcumin synergy is not apparent relative to 22.5 μm of ol/L
It improves.But in the case where resveratrol and curcumin are used separately, even if concentration reaches 25 μm of ol/L, the function and effect of the two
The sum of be also only 37.04 μm of ol/L (referring to Tables 1 and 2), and reduce half even with concentration when the two is used in combination,
Effect is also much better than the sum of effect being used alone.And when concentration reaches 25 μm of ol/L, the effect being used in combination can reach
93.23%, it is 2.5 times or more when being used separately, this technique effect is far beyond the expection model of those skilled in the art
It encloses.
Embodiment 3
By the optimization of embodiment 2, the present embodiment is selected in the case where fixed resveratrol concentration is 20 μm of ol/L, right
The ratio that resveratrol and curcumin are used in combination is advanced optimized.Respectively to resveratrol：Curcumin=1:1、1:
1.4、1:1.8、1:2.0、1:2.5 and 1:3 ratio optimization, measure every group of proliferation inhibition rate to human hepatoma HepG2 cell into
Measurement is gone, the results are shown in Table 5.
The joint component of 5 various concentration ratio of table is to HepG2Cell growth inhibiting effect (N=6)
As known from Table 5, as the concentration ratio of curcumin is continuously improved, resveratrol-curcumin administering drug combinations group is to people
Liver cancer HepG2The inhibiting rate of cell Proliferation is continuously improved on the whole.Wherein, in concentration ratio (resveratrol：Curcumin) it is 1:
When 2.5, inhibiting rate is up to 93.45%, is improved to 1 in concentration ratio:When 3, inhibiting rate is declined slightly.
Although the present invention is disclosed as above with preferred embodiment, it is not limited to the present invention, any to be familiar with this
The people of technology can do various changes and modification, therefore the protection of the present invention without departing from the spirit and scope of the present invention
Range should be subject to what claims were defined.
Claims (6)
1. a kind of having the composition for inhibiting hepatoma cell proliferation effect, which is characterized in that including using a concentration of 12.5 μm of ol/
The curcumin of the resveratrol and a concentration of 12.5 μm of ol/L-200 μm of ol/L of use of L-200 μm of ol/L.
2. according to claim 1 have the composition for inhibiting hepatoma cell proliferation effect, which is characterized in that the white lamb's-quarters
A concentration of 12.5 μm of ol/L-50 μm of ol/L of use of reed alcohol；A concentration of 12.5 μm of ol/L-100 μm of ol/ of use of the curcumin
L。
3. according to claim 2 have the composition for inhibiting hepatoma cell proliferation effect, which is characterized in that the white lamb's-quarters
The ratio between the concentration of reed alcohol and curcumin is：Resveratrol：Curcumin=1；(1-8).
4. according to claim 3 have the composition for inhibiting hepatoma cell proliferation effect, which is characterized in that the white lamb's-quarters
The ratio between the concentration of reed alcohol and curcumin is：Resveratrol：Curcumin=1；(1-2.5).
5. according to claim 3 have the composition for inhibiting hepatoma cell proliferation effect, which is characterized in that the white lamb's-quarters
A concentration of 12.5 μm of ol/L~30 μm ol/L of use of reed alcohol.
6. any compositions of claim 1-5 are preparing the application in treating medicines resistant to liver cancer.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810304163.7A CN108272780B (en) | 2018-04-08 | 2018-04-08 | Composition with effect of inhibiting liver cancer cell proliferation and application thereof |
Applications Claiming Priority (1)
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Citations (2)
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| CN105267187A (en) * | 2014-06-17 | 2016-01-27 | 优普生物科技股份有限公司 | Medicinal composition for treating fatty liver and preparation method and application thereof |
| CN106668060A (en) * | 2017-01-22 | 2017-05-17 | 朱定祥 | Anti-cancer preparation containing sulforaphane and selenium and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105267187A (en) * | 2014-06-17 | 2016-01-27 | 优普生物科技股份有限公司 | Medicinal composition for treating fatty liver and preparation method and application thereof |
| CN106668060A (en) * | 2017-01-22 | 2017-05-17 | 朱定祥 | Anti-cancer preparation containing sulforaphane and selenium and preparation method thereof |
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| Title |
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| QIN DU等: "Synergistic anticancer effects of curcumin and resveratrol in Hepa1-6 hepatocellular carcinoma cells", 《ONCOLOGY REPORTS》 * |
| 杜琴等: "白藜芦醇联合姜黄素对SMMC-7721肝癌细胞作用", 《中国实验方剂学杂志》 * |
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Effective date of registration: 20231008 Address after: 212310 Life Science Industrial Park, Development Zone, Danyang City, Zhenjiang City, Jiangsu Province Patentee after: Huashi Haolin (Danyang) Medical Technology Co.,Ltd. Address before: 116000, No. 15, 9th floor, 2nd floor, Zone A, Xinghai Square, Shahekou District, Dalian City, Liaoning Province Patentee before: DALIAN YUANGU KANGJIAN TECHNOLOGY CO.,LTD. |