CN111840350A - Artemisia selengensis alcohol extract with anti-inflammatory effect and preparation method and application thereof - Google Patents
Artemisia selengensis alcohol extract with anti-inflammatory effect and preparation method and application thereof Download PDFInfo
- Publication number
- CN111840350A CN111840350A CN202010624551.0A CN202010624551A CN111840350A CN 111840350 A CN111840350 A CN 111840350A CN 202010624551 A CN202010624551 A CN 202010624551A CN 111840350 A CN111840350 A CN 111840350A
- Authority
- CN
- China
- Prior art keywords
- extract
- alcohol
- alcohol extract
- inflammatory
- artemisia selengensis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/282—Artemisia, e.g. wormwood or sagebrush
-
- 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]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Rheumatology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pain & Pain Management (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Botany (AREA)
- Alternative & Traditional Medicine (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to the field of extraction of plant active components, in particular to an alcohol extract of artemisia selengensis with an anti-inflammatory effect, and a preparation method and application thereof. The invention adopts an ultrasonic extraction method, takes ethanol as an extraction solvent to obtain the alcohol extract of the cold fleabane, acts on an LPS-induced RAW264.7 cell model, and determines the anti-inflammatory property of the alcohol extract of the cold fleabane by detecting the influence of the extract on the cell survival rate and inflammatory medium.
Description
Technical Field
The invention relates to the field of extraction of plant active components, in particular to an alcohol extract of artemisia selengensis with an anti-inflammatory effect, and a preparation method and application thereof.
Background
Inflammation, a defense measure against infection or tissue damage, can occur in various tissues and organs of the human body, such as hepatitis, gastritis, pneumonia, etc. Wherein bacterial or viral infections, burns and cuts, strong acid and alkali attacks, and immune system disorders of the human body can cause inflammation of the body, and these factors are called inflammatory factors.
Artemisia frigida Willd is a perennial herb of Artemisia (Artemisia) of Compositae (Compositae). The total flavone extract of herba Artemisiae Scopariae has obvious antiinflammatory effect. However, the effective active components of the anti-inflammatory action of artemisia frigida and the action mechanism thereof are not clear.
Disclosure of Invention
The invention aims to provide an alcohol extract of artemisia selengensis with an anti-inflammatory effect.
The invention also aims to provide a preparation method of the artemisia selengensis alcohol extract.
The invention also aims to provide the application of the artemisia selengensis alcohol extract.
The alcohol extract of artemisia selengensis with anti-inflammatory effect is prepared by the following method,
pulverizing herba Artemisiae Scopariae and sieving;
adding 95% alcohol, placing into an ultrasonic extractor, stirring uniformly, and performing ultrasonic extraction with ultrasonic power of 500W, temperature of 50 deg.C for 30min, wherein the volume mass ratio of alcohol to Artemisia selengensis powder is 40:1 (mL/g);
after ultrasonic extraction, pouring the upper layer extracting solution into a centrifuge tube, collecting the centrifuged supernatant, and evaporating and concentrating to obtain the arteannuin extract.
The invention provides a preparation method of an alcohol extract of artemisia selengensis, which comprises the following steps:
pulverizing herba Artemisiae Scopariae and sieving;
adding 95% alcohol, placing into an ultrasonic extractor, stirring uniformly, and performing ultrasonic extraction with ultrasonic power of 500W, temperature of 50 deg.C for 30min, wherein the volume mass ratio of alcohol to Artemisia selengensis powder is 40:1 mL/g;
After ultrasonic extraction, pouring the upper layer extracting solution into a centrifuge tube, collecting the centrifuged supernatant, and evaporating and concentrating to obtain the arteannuin extract.
The invention also provides application of the artemisia selengensis alcohol extract in inhibiting inflammatory factors.
According to the use of the invention, the inflammatory factor is nitric oxide.
The invention provides application of the artemisia selengensis alcohol extract in preparing a preparation for inhibiting inflammatory factors nitric oxide.
The invention adopts an ultrasonic extraction method, takes ethanol as an extraction solvent to obtain the alcohol extract of the cold fleabane, acts on an RAW264.7 cell model induced by LPS, and determines the anti-inflammatory property of the alcohol extract of the cold fleabane by detecting the influence of the extract on the cell survival rate and inflammatory medium.
Drawings
FIG. 1 shows the effect of herbal alcohol extracts on cell survival;
FIG. 2 shows the effect of herbal alcohol extracts on cell survival;
FIG. 3 shows the effect of herbal alcohol extracts on cell survival;
FIG. 4 shows the effect of herbal alcohol extracts on cell survival;
FIG. 5 shows the result of ELISA kit for detecting the content of nitric oxide in the supernatant of RAW264.7 cells;
FIG. 6 shows the results of ELISA kit for detecting the content of interleukin-6 in the supernatant of RAW264.7 cells;
FIG. 7 shows the result of ELISA kit for detecting the content of interleukin-1 β in the supernatant of RAW264.7 cells;
FIG. 8 shows the result of ELISA kit for detecting TNF-alpha content in the supernatant of RAW264.7 cells.
Detailed Description
The invention takes 19 Chinese herbal medicines as raw materials, adopts an ultrasonic extraction method, takes ethanol as an extraction solvent, respectively obtains alcohol extracts of the 19 Chinese herbal medicines, and finds that different Chinese herbal medicines have different anti-inflammatory characteristics. Acting on RAW264.7 cell model induced by LPS, and determining the object of anti-inflammatory action research by detecting the influence of the extract on cell survival rate and inflammation mediators NO, IL-6, 1L-1 beta and TNF-alpha;
(2) determining effective active ingredients of the anti-inflammatory effect research object. Detecting the influence of the Chinese herbal medicine extract on the expression levels of an inflammation medium NO and inflammatory factors IL-6, IL-1 beta and TNF-alpha by using a mouse macrophage RAW264.7 inflammation model induced by LPS (low-cholesterol) to determine an effective anti-inflammatory Chinese herbal medicine extract; extracting the determined anti-inflammatory Chinese herbal medicine extract by using different organic solvents, investigating the influence of the extract on the cell survival rate and the expression amounts of inflammation mediator NO and inflammation factors IL-6, IL-1 beta and TNF-alpha, and determining effective components; fractionating by using a high performance liquid chromatography technology, examining the influence of each fraction on the cell survival rate and the expression levels of inflammation medium NO and inflammation factors IL-6, IL-1 beta and TNF-alpha, and determining effective fractions;
Example 1 preparation of an alcohol extract of Chinese herbal medicine
1. Pulverizing Chinese herbal medicine
Pulverizing herba Artemisiae Annuae, radix Pulsatillae, herba Artemisiae Annuae, cortex Dictamni Radicis, Oxytropis myriophylla, herba Viciae Fatuae, radix Saposhnikoviae, cortex astragali, folium astragali, Ganoderma stick, northeast flos Callicarpae Formosanae, rhizoma Atractylodis stem and leaf, rhizoma Atractylodis root and rhizome, herba Xanthii, radix Rumicis Japonici, folium Allii tuberosi, radix Rubiae, and exocarpium Benincasae with a miniature plant pulverizer, sieving with 60 mesh analytical sieve, and storing in sealed bag for use.
2. Preparation of alcohol extract
(1) Adding 800mL of 95% alcohol into an ultrasonic extractor, weighing 20g of artemisia annua coarse powder, putting the artemisia annua coarse powder into the ultrasonic extractor, uniformly stirring, and carrying out ultrasonic extraction under the ultrasonic conditions: the ultrasonic power is 500W, the temperature is 50 ℃, and the time is 30 min.
(2) After ultrasonic extraction, taking out a container, pouring an upper layer extracting solution into a centrifuge tube, centrifuging at 5000rpm for 20min, collecting or transferring a centrifuged supernatant into a rotary evaporation bottle, carrying out rotary concentration to about 5mL by using a rotary evaporator, transferring a liquid into a proper container by using a liquid transfer gun, wrapping the container with a preservative film, pricking a plurality of holes at the upper end of the container, putting the container into a refrigerator at minus 80 ℃, freezing the container into a solid, putting the solid into a freeze dryer, freeze-drying the solid into powder, taking down the preservative film, covering a cover of the centrifuge tube to serve as a sample to be detected, marking the sample, and putting the sample into the refrigerator at minus 80 ℃ for later use;
Preparing ethanol extracts of herba Artemisiae Scopariae, radix Pulsatillae, tarragon, cortex Dictamni Radicis, Oxytropis myriophylla, herba Viciae Fatuae, radix Saposhnikoviae, Astragalus membranaceus seed coat, Astragalus membranaceus leaf, Ganoderma stick, flos Rhododendri mollis, rhizoma Atractylodis stem and leaf, rhizoma Atractylodis root and rhizome, herba Xanthii, radix Rumicis Japonici, folium Allii Fistulosi, radix Rubiae, and exocarpium Pachyrhizi Erosi by the same method.
Weighing ethanol extract 10mg, placing in 1.5mL LEP tube, preparing into mother liquor with concentration of 10mg/mL in sterile environment, blowing and mixing with liquid transfer gun or inverting, dissolving, filtering with sterile 0.45 μm filter membrane to new sterile centrifuge tube, placing in-80 deg.C refrigerator, and storing for use.
EXAMPLE 2 anti-inflammatory action of herbal extracts
1. Effect of alcohol extracts on cell survival
The herb extracts of example 1 were determined to have an effect on cell survival. Each extract was applied to RAW264.7 cells at different concentrations (80. mu.g/mL, 100. mu.g/mL, 150. mu.g/mL, 200. mu.g/mL, 250. mu.g/mL) and the effect on cell viability was examined using CCK-8 reagent.
As can be seen from fig. 1 to 4, after the alcohol extracts of the 19 Chinese herbal medicines act on RAW264.7 cells, the eurotium cristatum alcohol extract, the saposhnikovia divaricata alcohol extract, the astragalus membranaceus leaf alcohol extract, the northeast rubus chingii alcohol extract, the cocklebur fruit stem leaf alcohol extract, the xanthium sibiricum alcohol extract, the rheum undulatum alcohol extract, the madder alcohol extract and the alcohol extract of the pachyrhizus peel all cause damage to the cells to different degrees.
2. Anti-inflammatory action of alcohol extract
Research on the anti-inflammatory effect of artemisia annua alcohol extract, artemisia frigida alcohol extract, pulsatilla chinensis alcohol extract, tarragon alcohol extract, cortex dictamni alcohol extract, echinocandin foliata alcohol extract, astragalus cochinchinensis alcohol extract, ganoderma lucidum fungus stick alcohol extract, atractylodes lancea rhizome root fibrous alcohol extract and Chinese chive alcohol extract without cytotoxicity.
The mouse mononuclear macrophage RAW264.7 induced by LPS is taken as a model, and a control group, an LPS group (LPS + culture medium) and an LPS + adding group are arranged. Treating RAW264.7 cells with the extract at a concentration of 100 μ g/mL for 2 hours, adding LPS at a final concentration of 1000 μ g/mL except for the control group, co-culturing for 24 hours, collecting cell supernatant, and detecting the content of anti-inflammatory factors.
And (3) detecting the influence of each alcohol extract on the expression quantity of NO, IL-6, IL-1 beta and TNF-alpha in cell supernatant by using an ELISA kit.
(2-1) influence of Chinese herbal medicine alcohol extract on NO content
As can be seen from fig. 5, the content of NO in the artemisia annua ethanol extract, the artemisia frigida ethanol extract, the radix pulsatillae ethanol extract and the artemisia tarragana ethanol extract treatment group is higher than that in the control group and lower than that in the LPS model group, which indicates that the artemisia annua ethanol extract, the artemisia frigida ethanol extract, the radix pulsatillae ethanol extract and the artemisia tarragana ethanol extract can inhibit the release of NO, which is an inflammation medium, and the capacity of inhibiting the release of NO is the artemisia annua ethanol extract, the artemisia frigida ethanol extract, the radix pulsatillae ethanol extract and the artemisia tarragana.
(2-2) influence of Chinese herbal medicine alcohol extract on IL-6 content
As can be seen from FIG. 6, compared with the LPS model group, the content of IL-6 is not significantly changed after the RAW264.7 cells are treated by the artemisia annua alcohol extract, the artemisia selengensis alcohol extract and the atractylodes rhizome root hair alcohol extract. After the radix pulsatillae alcohol extract and the alcohol extract of the ganoderma lucidum stick are used for treating RAW264.7 cells, the content of IL-6 in cell supernatant is not reduced, but is increased. The tarragon alcohol extract, the cortex dictamni alcohol extract, the echinocandin acutiloba alcohol extract, the astragalus membranaceus seed alcohol extract and the allium chinense alcohol extract obviously reduce the content of IL-6 in cell supernatant, which shows that the tarragon alcohol extract, the cortex dictamni alcohol extract, the astragalus membranaceus seed alcohol extract and the allium chinense alcohol extract can inhibit the release of IL-6 in the cell supernatant. The ability of inhibiting IL-6 is that the tarragon alcohol extract is white fresh pericarp alcohol extract and multi-leafy echinocandin bean alcohol extract and swertia chifolia alcohol extract is astragalus strictus seed extract.
(2-3) influence of Chinese herbal medicine alcohol extract on IL-1 beta content
As can be seen from FIG. 7, after the artemisia annua ethanol extract and the Chinese chive ethanol extract act on cells, the content of the inflammatory factor IL-1 beta is not reduced but increased, and the rest 8 ethanol extracts have no significant change compared with the LPS model group, which shows that the ethanol extracts of the 10 Chinese herbal medicines have no significant inhibition effect on the IL-1 beta in the cell supernatant.
(2-4) Effect of herbal alcohol extracts on TNF-alpha content
As can be seen from FIG. 8, the expression levels of TNF-alpha in the alcohol extracts of cortex Dictamni Radicis and folium Allii tuberosi are higher than those of the control group and LPS group, which indicates that both the alcohol extracts of cortex Dictamni Radicis and folium Allii tuberosi can inhibit the release and expression of TNF-alpha in the cell supernatant. The capability of inhibiting TNF-alpha is the Chinese chive alcohol extract, namely the alcohol extract of the white fresh pericarp. The other 8 alcohol extracts have no obvious inhibition effect on the release and expression of TNF-alpha.
In summary, as can be seen from fig. 5 to 8, the anti-inflammatory effects of the alcohol extracts are different, wherein the alcohol extract of artemisia selengensis can better inhibit the release of inflammatory mediator NO.
Claims (5)
1. The cold wormwood alcohol extract with anti-inflammatory effect is characterized by being prepared by the following method:
pulverizing herba Artemisiae Scopariae and sieving;
adding 95% alcohol, placing into an ultrasonic extractor, stirring uniformly, and performing ultrasonic extraction with ultrasonic power of 500W, temperature of 50 deg.C for 30min, wherein the volume mass ratio of alcohol to Artemisia selengensis powder is 40:1 mL/g;
after ultrasonic extraction, pouring the upper layer extracting solution into a centrifuge tube, collecting the centrifuged supernatant, and evaporating and concentrating to obtain the arteannuin extract.
2. The preparation method of the alcohol extract of artemisia selengensis is characterized by comprising the following steps:
Pulverizing herba Artemisiae Scopariae and sieving;
adding 95% alcohol, placing into an ultrasonic extractor, stirring uniformly, and performing ultrasonic extraction with ultrasonic power of 500W, temperature of 50 deg.C for 30min, wherein the volume mass ratio of alcohol to Artemisia selengensis powder is 40:1 mL/g;
after ultrasonic extraction, pouring the upper layer extracting solution into a centrifuge tube, collecting the centrifuged supernatant, and evaporating and concentrating to obtain the arteannuin extract.
3. Use of an alcohol extract of artemisia selengensis with anti-inflammatory effect as claimed in claim 1 for inhibiting inflammatory factors.
4. Use according to claim 3, wherein, according to the invention, said inflammatory factor is nitric oxide.
5. Use of an alcohol extract of artemisia selengensis with anti-inflammatory effect as claimed in claim 1 for the preparation of a preparation for inhibiting the inflammatory factor nitric oxide.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2020103748414 | 2020-05-07 | ||
| CN202010374841 | 2020-05-07 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111840350A true CN111840350A (en) | 2020-10-30 |
Family
ID=72484984
Family Applications (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010624537.0A Pending CN111700924A (en) | 2020-05-07 | 2020-07-01 | Artemisia frigida water extract and preparation method and application thereof |
| CN202010624540.2A Pending CN111686143A (en) | 2020-05-07 | 2020-07-01 | Artemisia frigida extract and preparation method and application thereof |
| CN202010624551.0A Pending CN111840350A (en) | 2020-05-07 | 2020-07-01 | Artemisia selengensis alcohol extract with anti-inflammatory effect and preparation method and application thereof |
Family Applications Before (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010624537.0A Pending CN111700924A (en) | 2020-05-07 | 2020-07-01 | Artemisia frigida water extract and preparation method and application thereof |
| CN202010624540.2A Pending CN111686143A (en) | 2020-05-07 | 2020-07-01 | Artemisia frigida extract and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (3) | CN111700924A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114152705B (en) * | 2021-12-06 | 2023-11-17 | 中国农业科学院饲料研究所 | HPLC fingerprint quality evaluation method for rhizoma atractylodis stem and leaf |
-
2020
- 2020-07-01 CN CN202010624537.0A patent/CN111700924A/en active Pending
- 2020-07-01 CN CN202010624540.2A patent/CN111686143A/en active Pending
- 2020-07-01 CN CN202010624551.0A patent/CN111840350A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 裴乐等: "蒙药小白蒿总黄酮物质提取工艺的优化", 《畜牧与饲料科学》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111700924A (en) | 2020-09-25 |
| CN111686143A (en) | 2020-09-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Rehmannia glutinosa: review of botany, chemistry and pharmacology | |
| AU2002348988B2 (en) | Composition comprising wenguanguo extracts, methods for preparing same and uses thereof | |
| JP5860577B2 (en) | Claudin production promoter and occludin production promoter | |
| AU2002348988A1 (en) | Composition comprising Wenguanguo extracts, methods for preparing same and uses thereof | |
| CN105031530A (en) | Anti-aging composition | |
| WO2019190161A1 (en) | Whole ginseng composition using ginseng roots, leaves and berries and method of preparing the same | |
| CN107998212A (en) | A kind of preparation method of glutinous rehmannia cyclic olefine ether terpinyl side like extractive | |
| CN110840924A (en) | Application of rhizoma atractylodis stem and leaf extract | |
| CN100482262C (en) | Herbal pharmaceutical compositions for treating immun logical disorders | |
| US6383525B1 (en) | Herbal compositions for treating immunological disorders | |
| CN111840350A (en) | Artemisia selengensis alcohol extract with anti-inflammatory effect and preparation method and application thereof | |
| CN113101310B (en) | Medicine for preventing and/or treating coronavirus infection and preparation method thereof | |
| CN103550129B (en) | Plant composite polysaccharide, Preparation Method And The Use | |
| CN1903241B (en) | Method for extraction and separation of pseudo-ginseng | |
| CN118078763A (en) | Granule for strengthening body resistance, eliminating dampness and resisting toxicity and preparation method thereof | |
| CN112494576A (en) | Oral liquid for treating fatty liver and alcoholic liver injury and preparation method thereof | |
| CN103316074B (en) | Medicine composite of halenia corni extract, astragalus extract and liquorice extract as well as preparation and application of medicine composite | |
| Nebedum et al. | Comparative effects of ethanolic extracts of Ficus carica and Mucuna pruriens leaves on haematological parameters in albino rats | |
| CN104435658B (en) | A kind of medicine for ischemia apoplexy prevention and treatment and preparation method thereof | |
| RU2642256C2 (en) | Application of composition in medical products or drug manufacture for prevention and treatment of leukopenia caused by radiation and chemotherapy | |
| CN1820780A (en) | Method for preparing Fuyankang extract | |
| CN113994980B (en) | Composition for preventing and treating pine wood nematode as well as preparation method and application thereof | |
| CN112386654B (en) | Traditional Chinese medicine composition, extract, preparation method and application thereof | |
| CN105535657A (en) | Traditional Chinese medicine composition for treating colon cancer | |
| CN106421715A (en) | Traditional Chinese medicinal composition for treating chronic colitis and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |