CN109288826A - Application of the aurantio-obtusin in preparation anti-inflammatory and pre- ageing prod against sunshine - Google Patents
Application of the aurantio-obtusin in preparation anti-inflammatory and pre- ageing prod against sunshine Download PDFInfo
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- CN109288826A CN109288826A CN201811335671.8A CN201811335671A CN109288826A CN 109288826 A CN109288826 A CN 109288826A CN 201811335671 A CN201811335671 A CN 201811335671A CN 109288826 A CN109288826 A CN 109288826A
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- obtusin
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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
- A61K31/12—Ketones
- A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
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Abstract
The invention discloses application of the aurantio-obtusin in preparation anti-inflammatory and pre- ageing prod against sunshine, it is target spot that the present invention induces RAW264.7 cell to discharge NO and IL-6 by LPS, aurantio-obtusin is demonstrated with good antiinflammation, it was demonstrated that aurantio-obtusin has a good application prospect in terms of preparing anti-inflammatory product；It is target spot that the present invention induces HaCaT cell to secrete IL-6 and IL-1 α by UVB, and demonstrating aurantio-obtusin has prevention light aging effect, it was demonstrated that aurantio-obtusin has a good application prospect in terms of preparing pre- ageing prod against sunshine.
Description
Technical field
The present invention relates to aurantio-obtusin fields, and specifically aurantio-obtusin is in preparation anti-inflammatory and pre- ageing prod against sunshine
Application.
Background technique
Cassia seed first recorded in Shennong's Herbal, be legume cassia (Cassia Obtusifolia L.) or it is small certainly
The dry mature seed of bright (Cassia tora L.), sweet in flavor, bitter, salty, cold nature, effect is heat-clearing improving eyesight, relax bowel and defecation.
Modern pharmacological studies have shown that cassia seed has lipid-loweringing, strengthen immunity, anti-oxidant, liver protection, cholagogue, relievings asthma, presses down
Bacterium, antiatherosclerosis and other effects.Aurantio-obtusin is ingredient exclusive in cassia seed, and character is that yellow needles fear quinones
Compound, the compound have that multiple biological activities are for example hypoglycemic, reducing blood lipid, antiestrogenic and inhibit collagen-induced blood small
Plate aggregation etc..
Currently, there has been no aurantio-obtusins for the report in terms of anti-inflammatory and pre- aging against sunshine.
Summary of the invention
The purpose of the present invention is to provide application of the aurantio-obtusin in preparation anti-inflammatory and pre- ageing prod against sunshine, with solution
Certainly the problems mentioned above in the background art.
To achieve the above object, the invention provides the following technical scheme:
Application of the aurantio-obtusin in preparation anti-inflammatory product.
As a further solution of the present invention: the aurantio-obtusin is when preparing anti-inflammatory product, the use of aurantio-obtusin
Amount is 5-20 μm of ol/L.
As a further solution of the present invention: the aurantio-obtusin is when preparing anti-inflammatory product, the use of aurantio-obtusin
Amount is 5 μm of ol/L.
As a further solution of the present invention: the aurantio-obtusin is when preparing anti-inflammatory product, the use of aurantio-obtusin
Amount is 20 μm of ol/L.
Aurantio-obtusin is preparing the application in pre- ageing prod against sunshine.
As a further solution of the present invention: the aurantio-obtusin is when preparing pre- ageing prod against sunshine, orange Cassia
The dosage of element is 10-20 μm of ol/L.
As a further solution of the present invention: the aurantio-obtusin is when preparing pre- ageing prod against sunshine, orange Cassia
The dosage of element is 10 μm of ol/L.
As a further solution of the present invention: the aurantio-obtusin is when preparing pre- ageing prod against sunshine, orange Cassia
The dosage of element is 20 μm of ol/L.
Compared with prior art, the beneficial effects of the present invention are:
First, the present invention induces RAW264.7 cell to discharge NO and IL-6 for target spot by LPS, demonstrates aurantio-obtusin
With good antiinflammation, it was demonstrated that aurantio-obtusin has a good application prospect in terms of preparing anti-inflammatory product；
Second, the present invention induces HaCaT cell to secrete IL-6 and IL-1 α for target spot by UVB, demonstrates aurantio-obtusin
With prevention light aging effect, it was demonstrated that aurantio-obtusin has a good application prospect in terms of preparing pre- ageing prod against sunshine.
Detailed description of the invention
Fig. 1 is influence diagram of the aurantio-obtusin to Raw264.7 cell viability.
Fig. 2 is the influence diagram that aurantio-obtusin induces LPS Raw264.7 cell secretion NO.
Fig. 3 is the influence diagram that aurantio-obtusin induces LPS Raw264.7 cell secretion IL-6.
Fig. 4 is the influence diagram that aurantio-obtusin secretes IL-6 to HaCaT cell.
Fig. 5 is the influence diagram that aurantio-obtusin secretes IL-1 α to HaCaT cell.
Specific embodiment
The technical solution of the patent is explained in further detail With reference to embodiment.
The RAW264.7 mouse macrophage of lipopolysaccharides (lipopolysaccharide, LPS) induction is that common research is scorching
The model of disease reaction.Macrophage lipopolysaccharides LPS stimulation under, can promote inflammatory factor such as nitric oxide (nitric oxide,
) and the secretion of interleukin-6 (interleukin 6) NO.The present invention discharges NO and IL-6 with LPS induction RAW264.7 cell
Target spot investigates the antiinflammation of aurantio-obtusin.
Skin aging includes natural aging and light aging, and wherein light aging removes the color, texture, elasticity, thickness for causing skin
Outside the variation of degree etc., in some instances it may even be possible to so that skin various benign or malignant tumours is occurred, influence serious.Skin photoage and UV irradiate
In close relations, ultraviolet B radiation UVB mainly acts on the keratinocyte in skin epidermis, skin appearance occurs above-mentioned
Variation and corresponding molecular biology change, cause the apoptosis of cell and secrete inflammatory factor IL-6 etc..HaCaT is common people
Epidermal keratinocyte system, the present invention investigate aurantio-obtusin using UVB induction HaCaT cell secretion IL-6 and IL-1 α as target spot
Prevent light aging effect.
Material and reagent: RAW264.7 cell strain is purchased from Chinese Academy of Sciences Shanghai cell institute；HaCaT is purchased from the Chinese Academy of Sciences
Shanghai cell institute, aurantio-obtusin, dexamethasone and DMSO (dimethyl sulfoxide) be purchased from Sigma company, DMEM high glucose medium,
Fetal calf serum FBS, pancreatin etc. are purchased from Gibco company, and CCK-8 is purchased from Shanghai past Biotechnology Co., Ltd；The cells such as IL-6
Factor ELISA kit is purchased from U.S. eBiosience company.
Drug solution preparing: the mother liquor that aurantio-obtusin is configured to 10mmol/L with DMSO is spare, with DMEM culture medium when dosing
It is diluted to the concentration of needs.
Cell culture: the DMEM high glucose medium culture of RAW264.7 cell 10%FBS containing mass fraction is placed in quality
Score 5%CO2, 37 DEG C, cultivate in the cell incubator of saturated humidity.Logarithmic growth phase is grown to cell to be passed on, 2~
3d is passed on 1 time.Cell after taking at least 3 generations is for testing.
The DMEM high glucose medium culture for being 10%FBS containing mass fraction of HaCaT cell, is placed in mass fraction 5%
CO2, 37 DEG C, cultivate in the cell incubator of saturated humidity.It grows to logarithmic growth phase to cell to be passed on, 2~3d passage 1
It is secondary.Cell after taking at least 3 generations is for testing.
The influence that drug and composition grow RAW264.7 cell: the RAW264.7 cell quality of logarithmic growth phase
Score is 0.25% pancreatin and mass fraction is after 0.02%EDTA digests 2min, pancreatin to be abandoned, with mass fraction 10%FBS's
It is acted in DMEM high glucose medium with pancreatin, gently blows and beats into single cell suspension, supernatant is abandoned in centrifugation, is resuspended with complete medium thin
Born of the same parents and after counting, adjustment cell suspension is to 8 × 105/ mL is seeded to 96 well culture plates, 37 DEG C, mass fraction by every 100 μ L of hole
5%CO2, cultivate for 24 hours under the conditions of saturated humidity, exhaust every hole supernatant, every hole is added 100 μ L DMEM high glucose mediums, random point
(contain mass fraction 5% for negative control group (the DMEM high glucose medium of the 0.1%DMSO containing mass fraction), toxicity control group
The DMEM high glucose medium of DMSO), medicine group；After relative medicine is added, every group sets 4 multiple holes, mass fraction 5%CO2、37
DEG C, cultivate for 24 hours under the conditions of saturated humidity after, CCK-8 is added by every 10 μ L of hole in 4h in advance, microplate reader detection 450nm place's suction after 4h
Shading value (A450).In Fig. 1 as can be seen that compared with blank control group, various concentration aurantio-obtusin is to RAW264.7 cell
Growth have no significant effect, i.e., aurantio-obtusin is acted in 5-20 μm of ol/L concentration acellular poison.
Inflammatory factor detection: the RAW264.7 cell mass fraction of logarithmic growth phase is 0.25% pancreatin and quality point
Number is after 0.02%EDTA digests 2min, to abandon pancreatin, is to make in 10%FBS DMEM high glucose medium with pancreatin with mass fraction
With gently blowing and beating into single cell suspension, supernatant is abandoned in centrifugation, and after cell is resuspended and is counted with complete medium, adjustment cell is outstanding
Liquid is to 8 × 105/ mL is seeded to 96 well culture plates, 37 DEG C, mass fraction 5%CO by every 100 μ L of hole2, train under the conditions of saturated humidity
Supporting 8h or so keeps cell adherent, exhausts every hole supernatant, and every hole is added 100 μ L serum-free DMEM high glucose mediums, is randomly divided into
Blank control group, LPS (1 μ g/mL) group, LPS+ positive controls (5 μm of ol/L Dex), LPS (1 μ g/mL)+medicine group are added
After relative medicine, every group sets 4 multiple holes, mass fraction 5%C02, 37 DEG C, cultivate for 24 hours under the conditions of saturated humidity after, take supernatant
Detection of the liquid for inflammatory factors such as NO, interleukin-6s (IL-6).As can be seen that various concentration aurantio-obtusin in Fig. 2
The NO secretion of the RAW264.7 cell of bacteria lipopolysaccharide (LPS) induction can be significantly inhibited, and is in dosage dependent effect.In Fig. 3
As can be seen that various concentration aurantio-obtusin can significantly inhibit the IL- of the RAW264.7 cell of bacteria lipopolysaccharide (LPS) induction
6 secretions, wherein the aurantio-obtusin antiphlogistic effects of 20 μM of concentration are best.
UVB light aging inflammatory factor detection: the Hacat cell mass fraction of logarithmic growth phase be 0.25% pancreatin and
Mass fraction is after 0.02%EDTA digests 2min, to abandon pancreatin, in mass fraction 10%FBS DMEM high glucose medium and pancreas
Enzyme effect gently blows and beats into single cell suspension, and supernatant is abandoned in centrifugation, and after cell is resuspended and is counted with complete medium, adjustment is thin
Born of the same parents' suspension is to 8 × 104/ mL is seeded to 24 well culture plates, 37 DEG C, mass fraction 5%CO by every 400 μ L of hole2, saturated humidity item
It is cultivated under part for 24 hours, exhausts every hole supernatant, after 500 μ L PBS of every hole addition are washed twice, added a small amount of PBS covering cell, use
UVB irradiation instrument irradiating cell (exposure intensity 8mJ/cm2), it has irradiated and has been washed once using PBS, blank group and UVB model group are added
The culture medium of aurantio-obtusin containing various concentration is added in 400 μ L culture mediums, administration group.After continuing culture 24 hours, Aspirate supernatant
Detection for IL-6 and IL-1 α.It can be seen that various concentration aurantio-obtusin in Fig. 4 and can significantly inhibit UVB and irradiate and cause
Hacat cell IL-6 secretion, and be in certain dosage dependent effect.As can be seen that the orange Cassia of 20 μm of ol/L in Fig. 5
Element can significantly inhibit the IL-1 α secretion of Hacat cell caused by UVB irradiates, and effective inhibiting rate reaches 22.4%.
In conclusion the RAW264.7 mouse macrophage induced using LPS, it has been found that aurantio-obtusin is to inflammation
The factor has good inhibiting effect.Secondly, we have detected what aurantio-obtusin was induced in UVB on the basis of anti-inflammatory result again
The protective effect of people's epidermal keratinocytes system HaCaT light aging, studies have shown that aurantio-obtusin can inhibit aging caused by UVB
Relevant inflammatory factors secretion.
It is obvious to a person skilled in the art that invention is not limited to the details of the above exemplary embodiments, Er Qie
In the case where without departing substantially from spirit or essential attributes of the invention, the present invention can be realized in other specific forms.Therefore, no matter
From the point of view of which point, the present embodiments are to be considered as illustrative and not restrictive, and the scope of the present invention is by appended power
Benefit requires rather than above description limits, it is intended that all by what is fallen within the meaning and scope of the equivalent elements of the claims
Variation is included within the present invention.Any reference signs in the claims should not be construed as limiting the involved claims.
In addition, it should be understood that although this specification is described in terms of embodiments, but not each embodiment is only wrapped
Containing an independent technical solution, this description of the specification is merely for the sake of clarity, and those skilled in the art should
It considers the specification as a whole, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art
The other embodiments being understood that.
Claims (6)
1. application of the aurantio-obtusin in preparation anti-inflammatory product.
2. application of the aurantio-obtusin according to claim 1 in preparation anti-inflammatory product, which is characterized in that described orange
The dosage of obtusin is 5-20 μm of ol/L.
3. application of the aurantio-obtusin according to claim 2 in preparation anti-inflammatory product, which is characterized in that described orange
The dosage of obtusin is 20 μm of ol/L.
4. aurantio-obtusin is preparing the application in pre- ageing prod against sunshine.
5. aurantio-obtusin according to claim 4 is preparing the application in pre- ageing prod against sunshine, which is characterized in that institute
The dosage for stating aurantio-obtusin is 10-20 μm of ol/L.
6. aurantio-obtusin according to claim 5 is preparing the application in pre- ageing prod against sunshine, which is characterized in that institute
The dosage for stating aurantio-obtusin is 20 μm of ol/L.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811335671.8A CN109288826B (en) | 2018-11-11 | 2018-11-11 | Application of aurantio-obtusin in preparing anti-inflammatory and anti-photoaging products |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811335671.8A CN109288826B (en) | 2018-11-11 | 2018-11-11 | Application of aurantio-obtusin in preparing anti-inflammatory and anti-photoaging products |
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| CN109288826A true CN109288826A (en) | 2019-02-01 |
| CN109288826B CN109288826B (en) | 2021-01-05 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111888346A (en) * | 2019-05-05 | 2020-11-06 | 上海医药集团股份有限公司 | Application of aurantio-obtusin |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101077341A (en) * | 2006-05-23 | 2007-11-28 | 成都中医药大学 | Application of aurantio-obtusifolin or its derivates in preparing hypolipidemic drug |
| CN101967090A (en) * | 2010-06-28 | 2011-02-09 | 南京泽朗农业发展有限公司 | Technology for extracting aurantio-obtusin |
| CN106581431A (en) * | 2016-12-10 | 2017-04-26 | 济南昊雨青田医药技术有限公司 | Medicine composition for preventing and treating amygdalitis |
| CN106727859A (en) * | 2016-12-10 | 2017-05-31 | 济南昊雨青田医药技术有限公司 | It is a kind of to treat pharmaceutical composition of pharyngitis and preparation method thereof |
-
2018
- 2018-11-11 CN CN201811335671.8A patent/CN109288826B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101077341A (en) * | 2006-05-23 | 2007-11-28 | 成都中医药大学 | Application of aurantio-obtusifolin or its derivates in preparing hypolipidemic drug |
| CN101967090A (en) * | 2010-06-28 | 2011-02-09 | 南京泽朗农业发展有限公司 | Technology for extracting aurantio-obtusin |
| CN106581431A (en) * | 2016-12-10 | 2017-04-26 | 济南昊雨青田医药技术有限公司 | Medicine composition for preventing and treating amygdalitis |
| CN106727859A (en) * | 2016-12-10 | 2017-05-31 | 济南昊雨青田医药技术有限公司 | It is a kind of to treat pharmaceutical composition of pharyngitis and preparation method thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111888346A (en) * | 2019-05-05 | 2020-11-06 | 上海医药集团股份有限公司 | Application of aurantio-obtusin |
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| CN109288826B (en) | 2021-01-05 |
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