CN104740566B - A kind of preparation for mitigating scytitis, preparation method and applications - Google Patents
A kind of preparation for mitigating scytitis, preparation method and applications Download PDFInfo
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Abstract
The present invention relates to a kind of preparations for mitigating scytitis, are prepared by two kinds of medicinal materials of Sculellaria barbata and rhizoma zingiberis, and the weight ratio of the Sculellaria barbata and rhizoma zingiberis is 1:0.9‐1.1.The invention further relates to a kind of applications of preparation for mitigating scytitis and preparation method thereof.The preparation of the mitigation scytitis of the present invention has advantageous effect that can be by scytitis degree up to mitigation to normal level；The preparation method of the preparation of the mitigation scytitis of the present invention has extraction efficiency is high, prepares preparation that is simple, at low cost, preparing can significantly mitigate scytitis.
Description
Technical field
The present invention relates to biomedicine technical field, more particularly to a kind of preparation for mitigating scytitis, preparation method and
It is applied.
Background technology
Sculellaria barbata (scientific name：Scutellaria barbata D.Don), also known as：Sun plant, pine needle tree peony, dragon tree peony,
Spun gold cuckoo, foreign purslane, heronsbill, Portulaca grandiflora, are horse lamiaceae labiatae scutellaria herbaceos perennials.《Correct Compendium of Materia Medica》
It records " this grass opens purple white flowers, careless aubergine, to peering leaf, use in seven Augusts ".The form is close with this kind.Traditional herbal medicine
In, it is a lot of with the kind that sun plant or " half " run after fame.《Hundred careless mirrors》It carries " various half have 72 kinds ".Zhao Xuemin exists《This
Careless detailed outline is picked up any lost article from the road》In recorded half branch of mouse tooth half, dog tooth half and protruding canine teeth etc., be the several plant of Crassulaceae Sedum.《In
Magnificent people's republic's pharmacopeia》Version in 1985, a sun plant recorded is this product.The product are the dry of labiate Sculellaria barbata
Dry herb.Summer, two season of autumn cauline leaf it is luxuriant when excavate, clean, dry.Have clearing heat and detoxicating, promoting blood circulation and removing blood stasis, swelling and pain relieving, anticancer it
Effect；It can appendicitis curing, hepatitis, stomachache, early liver cancer, lung cancer, cervix cancer, mastitis etc.；The treatment of pain furuncle is controlled in external application, bruise swells
The diseases such as pain.This medicine dosage among the people is larger.
Rhizoma zingiberis is the dry product of ginger.Ginger system perennial herb, high 40~100cm.Leaf 2 arranges, wire lanceolar, long by 15~
30cm, wide about 2cm, Glabrous.Scape bears from rhizome, high about 20cm；Spike is avette to oval；Bract pale green
Color, oval；Corolla yellow green, sliver lanceolar；The long round shape obovate of lip center sliver, it is short compared with corolla lobe, have pale purple
Vitta line and faint yellow spot；The micro- purple of stamen.Mainly produced in Sichuan, Guizhou and Luoping of Yunnan.Winter excavate, remove cauline leaf and
Fibrous root, clean dry or low temperature dry after, rhizoma zingiberis.Rhizoma zingiberis《Sheng Nong's herbal classic》It is classified as middle product, ginger《Mingyi Bielu》Row
For middle product.Tao Hongjing is called：It is " all to make rhizoma zingiberis method：Water logging three days, peeling are put in flowing water six, then more random peeling dries, puts porcelain
It makes three, is into cylinder." Li Shizhen (1518-1593 A.D.) meaning rhizoma zingiberis it is made with female ginger, using fair and clear solid person as good, it is all be used as medicine and Yi Pao use.Control waist
Cold, cold air, blood-breaking are ached in kidney, wind, logical extremities joint is removed, opens the vital organs of the human body, going the cold numbness of wind toxin and night mostly just.It controls and coughs, main middle benefit gas,
Cholera is more than, abdominal pain, dissipate-swelling expire cold dysentery and control blood and close.Patient is empty and cold, preferably adds and uses it.On the concocting method of rhizoma zingiberis, each family
Book on Chinese herbal medicine is recorded slightly different.But any is identical, is exactly " day exposure ".Other the step of, mainly have two point values that must discuss：
One, if peeling.《Mingyi Bielu》In i.e. have " water logging three days, peeling put in flowing water six, more scrape peeling, then dry, put
Make three, be into Porcelain Jar " record.It is known that CORTEX ZINGIBERIS RHIZOMAE nature and flavor are pungent, cool, spleen row water of advocating peace.The Ming Dynasty《Book on Chinese herbal medicine covers an aromatic plant metioned in ancient books》One
Have in book and " remove the peel heat, stay the record of Pi Liang ".Therefore, the property of the pungent-warm of CORTEX ZINGIBERIS RHIZOMAE and ginger is got along well, more with rhizoma zingiberis it is hot it
Property is not allowed, when not Wei two medicines, when ginger and rhizoma zingiberis are used as medicine, preferably remove the peel and be preferred.Modern pharmacology research is found, in all ginger
Processed product in, the volatile oil content in CORTEX ZINGIBERIS RHIZOMAE is minimum, this can also illustrate peeling whether the problem of.Secondly, if bubble.
《It is other to record name doctor》《Figure is through book on Chinese herbal medicine》Solarization is the record of rhizoma zingiberis after grade pharmaceutical texts have ginger bubble, some medicine and pharmacology works will
The step for be known as " water wine ".For Chinese Traditional Medicine theory, by bubble, the property of the pungent-warm of ginger can be weakened, make solution
Table power weakens, and work(is specially in.But the step for it is whether necessary, still up for further inquiring into.The ether extract and water of rhizoma zingiberis
Extract has apparent anti-inflammatory effect.Rat paw edema is without bright caused by rhizoma zingiberis preserved material and ginger ketenes gavage Carrageenan
Aobvious effect, ginger ketenes dosage, which increases by 1 times, then apparent anti-inflammatory effect, and intensity is weaker than the effect of indocin 5mg/kg.
The Chinese patent application of Application No. 200810030714 discloses a kind of prevention and treatment cardiovascular inflammation (poison
Element) Chinese medicine composition, by Sculellaria barbata, Radix Notoginseng, rhizoma atractylodis macrocephalae, Radix Codonopsis, Chinese cassia tree, cordate houttuynia, Radix Glycyrrhizae, the fruit of Chinese wolfberry, bittersweet, sealwort,
Jujube, the fleece-flower root, rhizoma zingiberis and oldenlandia diffusa are made, that is, disclose a kind of Chinese medicine composition containing Sculellaria barbata and ginger, have
The effect of clearing heat and detoxicating, promoting blood circulation and removing blood stasis, carbuncle that disappears dissipating bind, help consolidate has preferable prevention to the cardiovascular inflammation triggered by toxin
And therapeutic effect, to the hepatitis, pneumonia, gastritis, coronary heart disease, hypertension and the osteoproliferation that are triggered by cardiovascular inflammation and some
The diseases such as difficult and complicated cases also have good therapeutic effect；In addition, also there is the effect of certain for the internal organs tumour such as liver tumour, lung neoplasm.
The Chinese patent application of Application No. 200910028850 discloses a kind of medicine for treating acute gastritis, by Radix Salviae Miltiorrhizae,
Radix Paeoniae Alba, Sculellaria barbata, Radix Astragali, Poria cocos, dried orange peel, Radix Glycyrrhizae, green peel, rhizoma zingiberis, dragon's blood are made of raw material.Disclose raw material containing Sculellaria barbata and
The drug of rhizoma zingiberis is for treating acute gastritis, and with effect is good, beneficial effect of the curative for effect, high efficiency up to more than 80%
Fruit.
In the prior art without recording Sculellaria barbata and rhizoma zingiberis as raw material as preparation and for mitigating appointing for scytitis
What information and report.
The content of the invention
The technical problems to be solved by the invention are：A kind of mitigation dermatitis using Sculellaria barbata with rhizoma zingiberis as bulk pharmaceutical chemicals is provided
The preparation of disease, preparation method and applications.
In order to solve the above-mentioned technical problem, the technical solution adopted by the present invention is：
A kind of preparation for mitigating scytitis, is prepared by two kinds of medicinal materials of Sculellaria barbata and rhizoma zingiberis, the Sculellaria barbata and dry
The weight ratio of ginger is 1:0.9‐1.1.
Invention additionally discloses a kind of preparations for mitigating scytitis, are prepared including following medicinal material：Sculellaria barbata and rhizoma zingiberis,
The weight ratio of the Sculellaria barbata and rhizoma zingiberis is 1:0.9-1.1, the medicinal material respectively successively by alcohol is molten, concentration, it is lyophilized and
The extracting solution of back dissolving processing.
Invention additionally discloses a kind of preparation methods of the preparation of above-mentioned mitigation scytitis, and the medicinal material is placed respectively
It is impregnated in the ethyl alcohol for being 45-55% in volumetric concentration, acquisition soak, the volume ratio of the medicinal material and ethyl alcohol is respectively 1:
Soak when the soaking time is 65-80 small, is concentrated into medicinal material original weight amount, then vacuum freeze-drying by 15-25 under vacuum condition
To powdery, powder is obtained, the powder is subjected to back dissolving using pure water, by powder back dissolving made from medicinal material difference
To identical mass concentration, back dissolving liquid is obtained, back dissolving liquid made from medicinal material difference is mixed, obtains the preparation.
Invention additionally discloses a kind of preparation answering in mitigation scytitis product is prepared of above-mentioned mitigation scytitis
With.
The advantageous effect of the preparation of the mitigation scytitis of the present invention is：
(1) Sculellaria barbata and rhizoma zingiberis are prepared into pharmaceutical formulation for crude drug, so as to obtain the advantageous effect for mitigating scytitis；
(2) the more individual Sculellaria barbata of effect for the mitigation scytitis being formulated associated with Sculellaria barbata and rhizoma zingiberis or rhizoma zingiberis tool
There is significant enhancing, compared with the extracting solution generally containing Sculellaria barbata or rhizoma zingiberis, scytitis degree can be reached and mitigated to normal
It is horizontal.
The advantageous effect of the preparation method of the preparation of the mitigation scytitis of the present invention is：
(1) ethyl alcohol of the concentration for 45-55% is used, the volume ratio of the medicinal material and ethyl alcohol is respectively 1:15-25, the leaching
Technological means of the time for 65-80 when small is steeped, has the advantages that extraction efficiency is high；
(2) preparation method of the invention has the advantages that prepare simple, at low cost；
(3) preparation prepared can significantly mitigate scytitis.
Description of the drawings
Fig. 1 for no IL-17 binding signals chip scanning device scanning figure；
Fig. 2 is the chip scanning device scanning figure for having IL-17 binding signals；
Fig. 3 is the scanning figure of the experiment reactive tank of Radix Salviae Miltiorrhizae；
Fig. 4 is the scanning figure of the experiment reactive tank of Sculellaria barbata；
Fig. 5 is the scanning figure of the experiment reactive tank of rhizoma zingiberis；
Fig. 6 is the contrast table of the TNF-α mRNA level in-site of each group experiment；
Fig. 7 is the contrast table for the inhibitory action that different herbal extracts induce IL-17 TNF-α up-regulation；
Fig. 8 is the TNF-α mRNA level in-site table of each group experiment；
Fig. 9 is the HaCaT cell number scales of each experiment.
Specific embodiment
For the technology contents that the present invention will be described in detail, the objects and the effects, below in conjunction with embodiment and coordinate attached
Figure is explained.
The design of most critical of the present invention is:It is combined by Sculellaria barbata and rhizoma zingiberis, accelerates wound so as to which conspicuousness improves it
The speed of healing.
A kind of preparation of mitigation scytitis of the present invention, is prepared by two kinds of medicinal materials of Sculellaria barbata and rhizoma zingiberis, and described half
The weight ratio of branch lotus and rhizoma zingiberis is 1:0.9‐1.1.
Invention additionally discloses a kind of preparations for mitigating scytitis, are prepared including following medicinal material：Sculellaria barbata and rhizoma zingiberis,
The weight ratio of the Sculellaria barbata and rhizoma zingiberis is 1:0.9-1.1, the medicinal material respectively successively by alcohol is molten, concentration, it is lyophilized and
The extracting solution of back dissolving processing.
The advantageous effect of the preparation of the mitigation scytitis of the present invention is：
(1) Sculellaria barbata and rhizoma zingiberis are prepared into pharmaceutical formulation for crude drug, so as to obtain the advantageous effect for mitigating scytitis；
(2) the more individual Sculellaria barbata of effect for the mitigation scytitis being formulated associated with Sculellaria barbata and rhizoma zingiberis or rhizoma zingiberis tool
There is significant enhancing, compared with the extracting solution generally containing Sculellaria barbata or rhizoma zingiberis, scytitis degree can be reached and mitigated to normal
It is horizontal.
Further, the medicinal material further includes at least one of Radix Salviae Miltiorrhizae, stone reed, safflower, spina gleditsiae and dried pinellia, and described half
The weight ratio of branch lotus and rhizoma zingiberis is 1:1, it is preferred that the Sculellaria barbata and Radix Salviae Miltiorrhizae, stone reed, safflower, spina gleditsiae, the weight of dried pinellia
Ratio is respectively 1:1.
Invention additionally discloses a kind of preparation methods of the preparation of above-mentioned mitigation scytitis, and the medicinal material is placed respectively
It is impregnated in the ethyl alcohol for being 45-55% in volumetric concentration, acquisition soak, the volume ratio of the medicinal material and ethyl alcohol is respectively 1:
Soak when the soaking time is 65-80 small, is concentrated into medicinal material original weight amount, then vacuum freeze-drying by 15-25 under vacuum condition
To powdery, powder is obtained, the powder is subjected to back dissolving using pure water, by powder back dissolving made from medicinal material difference
To identical mass concentration, back dissolving liquid is obtained, back dissolving liquid made from medicinal material difference is mixed, obtains the preparation.
The advantageous effect of the preparation method of the preparation of the mitigation scytitis of the present invention is：
(1) ethyl alcohol of the concentration for 45-55% is used, the volume ratio of the medicinal material and ethyl alcohol is respectively 1:15-25, the leaching
Technological means of the time for 65-80 when small is steeped, has the advantages that extraction efficiency is high；
(2) preparation method of the invention has the advantages that prepare simple, at low cost；
(3) preparation prepared can significantly mitigate scytitis.
Further, the temperature of the concentration is 50-70 DEG C.
Further, the concentration of the ethyl alcohol is 50%, and the volume ratio of the medicinal material and ethyl alcohol is 1:20, during the immersion
Between for 72 it is small when.
Invention additionally discloses a kind of preparation answering in mitigation scytitis product is prepared of above-mentioned mitigation scytitis
With.
Further, the mitigation scytitis product is drug, health products, food, food additives or cosmetic products.
The embodiment of the present invention one is：
A kind of preparation of mitigation scytitis of the present embodiment is prepared by two kinds of medicinal materials of Sculellaria barbata and rhizoma zingiberis, described
The weight ratio of Sculellaria barbata and rhizoma zingiberis is 1:0.9.
A kind of preparation method of the preparation of above-mentioned mitigation scytitis of the present embodiment, the medicinal material is respectively placed in
Concentration is to be impregnated in 45% ethyl alcohol, acquisition soak, and the volume ratio of the medicinal material and ethyl alcohol is respectively 1:15, the leaching
When the bubble time is 65 small, soak is concentrated into medicinal material original weight amount under vacuum condition, then vacuum freeze-drying obtains powdery to powdery
The powder is carried out back dissolving by object using pure water, by the medicinal material respectively made from powder back dissolving to identical mass concentration,
Back dissolving liquid is obtained, back dissolving liquid made from medicinal material difference in same volume ratio is mixed, obtains the preparation.The concentration
Temperature be 50 DEG C.
The embodiment of the present invention two is：
Only " weight ratio of the Sculellaria barbata and rhizoma zingiberis is 1:1, the concentration of the ethyl alcohol is 50%, the medicinal material and second
The volume ratio of alcohol is respectively 1:20, the soaking time for 72 it is small when, the temperature of the concentration is 60 DEG C " different, remaining and reality
It is identical to apply example one.
The embodiment of the present invention three is：
Only " weight ratio of the Sculellaria barbata and rhizoma zingiberis is 1:1.1, the concentration of the ethyl alcohol is 55%, the medicinal material with
The volume ratio of ethyl alcohol is respectively 1:25, the soaking time for 80 it is small when, the temperature of the concentration is 70 DEG C " it is different, remaining with
Embodiment one is identical.
The embodiment of the present invention four is：
A kind of preparation of mitigation scytitis of the present embodiment, is prepared including following medicinal material：Sculellaria barbata and rhizoma zingiberis, institute
The weight ratio for stating Sculellaria barbata and rhizoma zingiberis is 1:, 0.9, the medicinal material respectively successively by alcohol is molten, concentration, at lyophilized and back dissolving
The extracting solution of reason.The medicinal material further includes at least one of Radix Salviae Miltiorrhizae, stone reed, safflower, spina gleditsiae and dried pinellia.
A kind of preparation method of the preparation of above-mentioned mitigation scytitis of the present embodiment, the medicinal material is respectively placed in
Concentration is to be impregnated in the ethyl alcohol of 45-55%, acquisition soak, and the volume ratio of the medicinal material and ethyl alcohol is respectively 1:15-25,
The soaking time for 65-80 it is small when, soak is concentrated into medicinal material original weight amount under vacuum condition, then vacuum freeze-drying is to powder
Shape obtains powder, and the powder is carried out back dissolving using pure water, by powder back dissolving made from medicinal material difference to phase
Homogenous quantities concentration obtains back dissolving liquid, and back dissolving liquid made from medicinal material difference in same volume ratio is mixed, obtains the system
Agent.The temperature of the concentration is 50-70 DEG C.
The embodiment of the present invention five is：
Only " mitigate the preparation of scytitis, be prepared including following medicinal material：Sculellaria barbata and rhizoma zingiberis, the Sculellaria barbata and
The weight ratio of rhizoma zingiberis is 1:1, the medicinal material respectively passes through molten alcohol, concentration, the extracting solution of lyophilized and back dissolving processing successively.Institute
State medicinal material and further include Radix Salviae Miltiorrhizae, stone reed, safflower, spina gleditsiae and dried pinellia " it is different, remaining is identical with example IV.
The embodiment of the present invention six is：
Only " mitigate the preparation of scytitis, be prepared including following medicinal material：Sculellaria barbata and rhizoma zingiberis, the Sculellaria barbata and
The weight ratio of rhizoma zingiberis is 1:1.1, the medicinal material respectively passes through molten alcohol, concentration, the extracting solution of lyophilized and back dissolving processing successively.
The medicinal material further includes any one in Radix Salviae Miltiorrhizae, stone reed, safflower, spina gleditsiae and dried pinellia " different, remaining and example IV phase
Together.
The embodiment of the present invention seven is：
Only " mitigate the preparation of scytitis, be prepared including following medicinal material：Sculellaria barbata and rhizoma zingiberis, the Sculellaria barbata and
The weight ratio of rhizoma zingiberis is 1:1, the medicinal material respectively passes through molten alcohol, concentration, the extracting solution of lyophilized and back dissolving processing successively.Institute
State medicinal material and further include arbitrary more than two or three in Radix Salviae Miltiorrhizae, stone reed, safflower, spina gleditsiae and dried pinellia " different, remaining and implementation
Example four is identical.
Effect analysis
Embodiment one to the medicinal material described in embodiment seven using containing the Chinese-herbal medicine chip of the medicinal material as raw material sources,
All Chinese-herbal medicine chips derive from Yunnan Paiyao Group Corp., Ltd.
Table 1 is the Category List of the Chinese-herbal medicine chip hybridized with Interleukin-17.All Chinese-herbal medicine chips are by Yunnan
Baiyao Group Plc provides.The biotinylated Interleukin-17 (IL-17) that 4ng concentration is 200ng/mL
It is separately added into table 1 in the experiment reactive tank of 16 kinds of (each each 10) Chinese-herbal medicine chips and is hybridized.Simultaneously the solution of equivalent
It is separately added into table 1 in the blank control reactive tank of 16 kinds of (each each 10) Chinese-herbal medicine chips.Hybridize 3 it is small when after, use phosphate
Buffered saline embathes chip 4 times, 15 minutes every time.Add 20-200ng STREPTAVIDIN Cy5 CONJUGATE (Life
Technologies) (10 μ g/mL) carries out secondary hybridization.Hybridize 3 it is small when after, embathe chip 4 times with phosphate buffered saline (PBS), often
Secondary 15 minutes.Finally using chip scanning device (GenePix 4100A) in the item of 600nm excitation wavelengths and 670nm launch wavelengths
It is scanned under part, obtains Fig. 1 and Fig. 2.
Table 1
| Sequence number | Title | Sequence number | Title | Sequence number | Title | Sequence number | Title |
| 1 | Radix Salviae Miltiorrhizae | 5 | Safflower | 9 | Fructus toosendan | 13 | Radix Polygalae |
| 2 | The radix paeoniae rubrathe | 6 | Spina gleditsiae | 10 | Radix scutellariae | 14 | Cicada slough |
| 3 | The seed of cowherb | 7 | Dandelion | 11 | Sculellaria barbata | 15 | Fructus Forsythiae |
| 4 | Stone reed | 8 | Field pennycress | 12 | Dried pinellia | 16 | Rhizoma zingiberis |
Fig. 1 is the chip scanning device scanning figure of no IL-17 binding signals, and Fig. 1 tops are the scanning of blank control reactive tank
Figure, Fig. 1 lower sections are the scanning figure of experiment reactive tank, and as shown in Figure 1, the experiment reactive tank of Fig. 1 does not generate IL-17 and combines letter
Number, that is, the medicinal material tested in reactive tank cannot be combined with IL-17.
Fig. 2 is the chip scanning device scanning figure for having IL-17 binding signals, is the scanning of blank control reactive tank above Fig. 2
Scheme, be the scanning figure of experiment reactive tank below Fig. 2, circled is that target combination is believed in the positive control for be printed on biotin in Fig. 2
Number, the generation of experiment false positive phenomenon can be excluded.As shown in Figure 2, the experiment reactive tank of Fig. 2 generates IL-17 binding signals, i.e.,
Medicinal material in experiment reactive tank can be combined with IL-17.
From 16 kinds of Chinese-herbal medicine chips, Sculellaria barbata, rhizoma zingiberis, Radix Salviae Miltiorrhizae, stone reed, safflower, the experiment reactive tank of spina gleditsiae and dried pinellia
Scanning figure as shown in Fig. 2, having binding signal, above-mentioned 7 kinds of medicinal materials have the property that can be combined with IL-17, remaining 9 kinds of grass
For the scanning figure of the experiment reactive tank of medicine core piece as shown in Figure 1, not having binding signal, above-mentioned 9 kinds of medicinal materials do not have energy and IL-
17 properties combined.In the scanning figure for the medicinal material that 9 kinds can be combined with VEGF, with the combination of Sculellaria barbata and rhizoma zingiberis and IL-17 most
To be apparent, reference can be made to Fig. 3-5.
Fig. 3 is the scanning figure of the experiment reactive tank of Radix Salviae Miltiorrhizae；Fig. 4 is the scanning figure of the experiment reactive tank of Sculellaria barbata；Fig. 5 is dry
The scanning figure of the experiment reactive tank of ginger.
Immortalization epidermal cell system (HaCaT) cell is pressed 2 × 104/ hole kinds in 6 porocyte culture plates.Second day, in
Radix Salviae Miltiorrhizae, stone reed, safflower, spina gleditsiae, Sculellaria barbata, dried pinellia and the herbal extract of rhizoma zingiberis are separately added into culture solution, culture 24 is small
When；Concentration is separately added into the culture solution of IL-17 and 10 μM of epithelical cell growth factor (EGF) containing 10ng/mL as 10 μ
Radix Salviae Miltiorrhizae, stone reed, safflower, spina gleditsiae, Sculellaria barbata, dried pinellia and the herbal extract of rhizoma zingiberis of g/mL, when culture 24 is small；It sets simultaneously
The experiment of the IL-17 of 10ng/mL is only added in one culture solution and one does not add the blank control of any reagent and medicinal material experiment.
16 groups of experiments are carried out altogether.Extract RNA according to the step of offer in RNAzol instrument cases, and using MMLV reverse transcriptases and
RNA reverse transcriptions into cDNA, are used tumor necrosis factor-alpha (TNF- by Oligo dT (poly thymidine, T repeat oligonucleotides)
α) exclusive primer 5'-AGTGACAAGCCTGTAGCC-3' and 5'-AGGTTGACTTTCTCCTGG-3' carry out PCR amplification (PCR
Amplification is：94 DEG C 1 minute；60 DEG C 1 minute；72 DEG C 1 minute, 30 cycle), using the gene expression dose of GAPDH for weigh
Figureofmerit obtains Fig. 6 and Fig. 7.
Fig. 6 is the contrast table of the TNF-α mRNA level in-site of each group experiment；In Fig. 6, ordinate is TNF-α mRNA (* radixes),
The experiment that abscissa is corresponding in turn to is：Blank control, IL-17, Radix Salviae Miltiorrhizae, IL-17+ Radix Salviae Miltiorrhizaes, stone reed, IL-17+ stone reeds, safflower,
IL-17+ safflowers, spina gleditsiae, IL-17+ spina gleditsiaes, Sculellaria barbata, IL-17+ Sculellaria barbatas, dried pinellia, IL-17+ dried pinellias, rhizoma zingiberis and IL-
17+ rhizoma zingiberis.
Fig. 7 is the contrast table for the inhibitory action that different herbal extracts induce IL-17 TNF-α up-regulation；In Fig. 7, indulge and sit
The inhibitory action of IL-17 induction TNF-α up-regulations is designated as, the extracting solution that abscissa is corresponding in turn to addition is：Radix Salviae Miltiorrhizae, stone reed, safflower,
Spina gleditsiae, Sculellaria barbata, dried pinellia and rhizoma zingiberis.
From Fig. 6 and Fig. 7, Sculellaria barbata and rhizoma zingiberis extracting solution induce IL-17 the inhibitory action of TNF-α up-regulation than remaining
The extracting solution of five kinds of herbal medicine that can be combined with IL-17 is height.
So carrying out compatibility using Sculellaria barbata and rhizoma zingiberis extracting solution, and Compatibility Results are tested with identical embodiment step.
That is, immortalization epidermal cell system (HaCaT) cell is pressed 2 × 104/ hole kinds in 6 porocyte culture plates.Second day, in culture
The herbal extract of Sculellaria barbata, rhizoma zingiberis and Sculellaria barbata+rhizoma zingiberis is separately added into liquid, when culture 24 is small；In containing 10ng/mL's
Sculellaria barbata, the rhizoma zingiberis that concentration is 10 μ g/mL are separately added into the culture solution of IL-17 and 10 μM of epithelical cell growth factor (EGF)
With the herbal extract of Sculellaria barbata+rhizoma zingiberis, culture 24 it is small when；The IL-17 that 10ng/mL is only added in one culture solution is set simultaneously
Experiment and one plus the blank control of any reagent and medicinal material experiment.8 groups of experiments are carried out altogether.According to RNAzol instrument cases
The step of interior offer, extracts RNA, and utilizes MMLV reverse transcriptases and Oligo dT (poly thymidine, T repeat oligonucleotides)
RNA reverse transcriptions into cDNA, the exclusive primer 5'-AGTGACAAGCCTGTAGCC-3' of tumor necrosis factor-alpha (TNF-α) is used
Carrying out PCR amplification with 5'-AGGTTGACTTTCTCCTGG-3', (PCR amplification parameter is：94 DEG C 1 minute；60 DEG C 1 minute；72℃1
Minute, 30 Xun Huans), using the gene expression dose of GAPDH as measurement index, obtain Fig. 8.
Fig. 8 is the TNF-α mRNA level in-site table of each group experiment；In Fig. 8, ordinate be TNF-α mRNA (* radixes), abscissa
The experiment being corresponding in turn to is：Blank control, IL-17, Sculellaria barbata, IL-17+ Sculellaria barbatas, rhizoma zingiberis, IL-17+ rhizoma zingiberis, Sculellaria barbata+dry
Ginger and IL-17+ Sculellaria barbatas+rhizoma zingiberis.
As shown in Figure 8, the formula for the back dissolving extracting solution of Sculellaria barbata and rhizoma zingiberis being mixed and being prepared by same components, energy
IL-17 induction TNF-α up-regulation effects are reduced to the horizontal almost identical of blank control.
HaCaT cells are pressed 6 × 103/ hole kinds in 96 porocyte culture plates.It second day, is separately added into culture solution
10ng/mL IL-17,1 μ g/mL IL-17,10 μM of EGF, 10 μ g/mL Sculellaria barbatas ,+1 μ g/mL IL-17 of 10 μ g/mL Sculellaria barbatas,
10 μ g/mL rhizoma zingiberis ,+1 μ g/mL IL-17 of 10 μ g/mL rhizoma zingiberis ,+10 μ g/mL rhizoma zingiberis of 10 μ g/mL Sculellaria barbatas, 5 μ g/mL Sculellaria barbatas+5
+ 1 μ g/mLIL-17 extracting solutions of μ g/mL rhizoma zingiberis, when culture 24 is small.Simultaneously one group of blank control is set to test.Take out 96 hole cells
Culture plate makees mtt assay test experience.
MTT test experience steps are as follows：The MTT reagents of 10 μ L are added in, the ultimate density of every hole milliliter is allowed to be 0.5mg/mL.
96 porocyte culture plates are put into 3 hours in 37 DEG C of incubators, after taking-up, cell culture fluid is siphoned away and adds in 100 μ
L dimethyl alums (DMSO) are protected from light down and shake up 15 minutes, detect the absorptivity under 570nm.According to standard curve absorptivity
It is scaled HaCaT cell quantities to make comparisons, obtains Fig. 9.
Fig. 9 is the HaCaT cell number scales of each experiment, and in Fig. 9, ordinate is HaCaT cell quantities (* 103), abscissa
Corresponding experiment is followed successively by：Blank control, 10ng/mL IL-17,1 μ g/mL IL-17,10 μM of EGF, 10 μ g/mL Sculellaria barbatas,
+ 1 μ g/mL IL-17 of 10 μ g/mL Sculellaria barbatas, 10 μ g/mL rhizoma zingiberis ,+1 μ g/mL IL-17 of 10 μ g/mL rhizoma zingiberis, 10 μ g/mL Sculellaria barbatas
+ 10 μ g/mL rhizoma zingiberis ,+1 μ g/mL IL-17 of+5 μ g/mL rhizoma zingiberis of 5 μ g/mL Sculellaria barbatas.
As shown in Figure 9, match somebody with somebody and can incite somebody to action what the back dissolving extracting solution of Sculellaria barbata and rhizoma zingiberis was mixed and prepared by same components
Human keratinocyte's layer epidermal cell cel l proliferation is down to the horizontal almost identical of blank control.
Mitigate in conclusion the preparation provided by the invention for mitigating scytitis has that scytitis degree can be reached to just
The flat advantageous effect of ordinary water；The preparation method of the preparation of the mitigation scytitis of the present invention have extraction efficiency is high, it is simple to prepare,
Preparation that is at low cost, preparing can significantly mitigate scytitis.
The foregoing is merely the embodiment of the present invention, are not intended to limit the scope of the invention, every to utilize this hair
The equivalents that bright specification and accompanying drawing content are made directly or indirectly are used in relevant technical field, similarly include
In the scope of patent protection of the present invention.
Claims (6)
1. a kind of preparation for mitigating scytitis, which is characterized in that be prepared by two kinds of medicinal materials of Sculellaria barbata and rhizoma zingiberis, described half
The weight ratio of branch lotus and rhizoma zingiberis is 1:0.9-1.1.
2. a kind of preparation for mitigating scytitis, which is characterized in that be prepared by following medicinal material：Sculellaria barbata and rhizoma zingiberis, it is described
The weight ratio of Sculellaria barbata and rhizoma zingiberis is 1:0.9-1.1, the medicinal material respectively successively by alcohol is molten, concentration, at lyophilized and back dissolving
Reason, obtains extracting solution.
3. the preparation method of the preparation of the mitigation scytitis described in a kind of claim 1-2 any one, which is characterized in that will
The medicinal material, which is respectively placed in the ethyl alcohol that volumetric concentration is 45-55%, to be impregnated, and obtains soak, the medicinal material and ethyl alcohol
Volume ratio be 1:Soak when the soaking time is 65-80 small, is concentrated into medicinal material original weight by 15-25 under vacuum condition
Amount, then vacuum freeze-drying obtain powder, the powder are carried out back dissolving using pure water, the medicinal material is made respectively to powdery
The powder back dissolving obtained obtains back dissolving liquid, back dissolving liquid made from medicinal material difference is mixed, obtains institute to identical mass concentration
State preparation.
4. the preparation method of the preparation according to claim 3 for mitigating scytitis, which is characterized in that the temperature of the concentration
It spends for 50-70 DEG C.
5. the preparation method of the preparation according to claim 3 for mitigating scytitis, which is characterized in that the body of the ethyl alcohol
Product concentration is 50%, and the volume ratio of the medicinal material and ethyl alcohol is 1:20, when the soaking time is 72 small.
6. a kind of preparation of the mitigation scytitis described in claim 1-2 any one is preparing the drug of mitigation scytitis
Or the application in cosmetic products.
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