CN105434567B - A kind of Chinese medicine composition and preparation method thereof for treating rheumatoid arthritis - Google Patents
A kind of Chinese medicine composition and preparation method thereof for treating rheumatoid arthritis Download PDFInfo
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Abstract
The present invention relates to a kind of Chinese medicine composition and preparation method thereof for treating rheumatoid arthritis, Chinese medicine composition is made of following extract: attached art polysaccharide 15~35%, atractylodes lactone 23~43%, Radix Aconiti Lateralis Preparata alkaloid 30~50%.The preparation method is as follows: 1) preparation of attached art polysaccharide: weighing monkshood and Rhizoma Atractylodis Macrocephalae, it is extracted with petroleum ether, ethyl alcohol decontamination is used again, and then the dregs of a decoction are extracted with water, obtain concentrated medicament, again by concentrated medicament, add alcohol precipitation after dehydrated alcohol, then after removing isolating protein, again alcohol precipitation, then it is successively washed with dehydrated alcohol, acetone, vacuum drying obtains attached art polysaccharide；2) preparation of atractylodes lactone and Radix Aconiti Lateralis Preparata alkaloid is centrifuged off insoluble matter after being flowed back with ethanol water bath, then adds to macroporous resin column upper end and successively uses pure water, ethanol elution, collect eluent, vacuum drying obtains.The present invention is well used in terms of preparing drugs for rheumatoid arthritis.
Description
Technical field
The present invention relates to the field of Chinese medicines, and in particular to the active principle Radix Aconiti Lateralis Preparata total alkaloids extracted from monkshood, from Rhizoma Atractylodis Macrocephalae
The composition and its preparation of the active principle atractylodes lactone of middle extraction and the attached art polysaccharide of the active principle extracted from monkshood Rhizoma Atractylodis Macrocephalae
Method.
Background technique
Rheumatoid arthritis (rheumatoid arthritis, RA) is a kind of common with progressive articular cartilage and bone
The autoimmune disease for damaging the chronic inflammation being characterized, finally can lead to joint deformity and function is lost, and disability rate is high.
It is shown according to latest survey, the disease incidence of China's rheumatoid arthritis is very high, and number of patients is about 30,000,000 or more, in adult
Disease incidence may be up to 2% or so.It is the one kind of " rheumatism ", traditional Chinese medicine in the traditional Chinese medical theory system of rheumatoid arthritis in ancient times
It is with a history of thousands of years to treat rheumatism, early in " Treatise on Febrile and Miscellaneous Disease " of Huangdi's Internal Classics and Zhang Zhongjing, " Synopsis Golden Chamber ", just
The discussion of system has been carried out to the cause of disease of rheumatism, the interpretation of the cause, onset and process of an illness, sick position, syndrome, prognosis etc.." monkshood-Rhizoma Atractylodis Macrocephalae " medicine is to coming from
The classical prescription of Zhang Zhongjing and modern clinic are used to treat the herbal pair of rheumatoid arthritis.Monkshood, it is warming channel and expelling cold to pass through
12 warps, Rhizoma Atractylodis Macrocephalae eliminating dampness, that closes then has dispellieg cold and dampness, and degrading the channel, the function of stopping numbness pain is the characteristic compatibility of the side's Zhong Jing treatment " bi Zheng "
One of, the two complementation is to use, and embodies the principles of formulating prescriptions of differential diagnosis in tcm.Studies have shown that monkshood and Rhizoma Atractylodis Macrocephalae compatibility can effectively inhibit
The secondary arthroncus of adjuvant arthritis rats, can significantly reduce the level of adjuvant arthritis rats serum NO level and TNF-α.
And the research of monkshood Rhizoma Atractylodis Macrocephalae medicine pair is essentially consisted at present, by experimental animal model, carry out compatibility of drugs front and back
Pharmacological effect and pharmaceutical chemistry research so that it is had scientific basis to carry out scientific evidence or verifying to traditional compatibility
Or reasonable experimental basis is provided for clinical application come developing new drug by the experimental study of compatibility.In addition, also by not year-on-year
The dosage ratio research of example, to disclose its dose ratio for playing the best use.The research side of the above monkshood Rhizoma Atractylodis Macrocephalae Compatibility Law
Method rests essentially within Chinese medicine itself generally, and itself is a complicated systems for Chinese medicine, have complicated chemical component
And drug action, therefore using the main active substances in Chinese medicine compound prescription as research object, study the proportion compatibility of its active material simultaneously
The secondary development for carrying out drug based on this is more reasonable.
Summary of the invention
The first purpose of the invention is to provide a kind of Chinese medicine composition for treating rheumatoid arthritis, of the invention second
A purpose is to provide a kind of preparation method of Chinese medicine composition for treating rheumatoid arthritis.
In order to realize above-mentioned first purpose, present invention employs technical solutions below:
A kind of Chinese medicine composition for treating rheumatoid arthritis, is made of the extract of following parts by weight: attached art polysaccharide 15
~35%, atractylodes lactone 23~43%, Radix Aconiti Lateralis Preparata alkaloid 30~50%.
Preferably, the ratio of the attached art polysaccharide is 25.5%, and the ratio of atractylodes lactone is 33.0%, and monkshood is raw
The ratio of alkaloids is 41.5%.
Preferably, the attached art polysaccharide is extracted by monkshood and Rhizoma Atractylodis Macrocephalae, and the atractylodes lactone is extracted by Rhizoma Atractylodis Macrocephalae, described
Radix Aconiti Lateralis Preparata alkaloid is extracted by RADIX ACONITI LATERALIS PREPARATA.
In order to realize above-mentioned second purpose, present invention employs technical solutions below:
A kind of preparation method of Chinese medicine composition that treating rheumatoid arthritis, comprising the following steps:
1) preparation of attached art polysaccharide: weighing monkshood and Rhizoma Atractylodis Macrocephalae 1000g, and wherein monkshood dosage is 3 times of Rhizoma Atractylodis Macrocephalae, is added
The petroleum ether that 6 times of amount volumes and boiling point are 60 DEG C -90 DEG C extracts 2 times, after extracting 3 hours every time, collects the dregs of a decoction and 80% second is added
Alcohol, 100 DEG C water-bath refluxing extraction 2 times, 3 hours every time, 8 times of amount solvents every time, remove fat in medicinal material, pigment, monosaccharide and
Oligosaccharides；After collecting 60 DEG C of vacuum drying 12h of the dregs of a decoction, add 100 DEG C of water of 10 times of amounts to extract 3 hours, extracts altogether twice, merge and extract
Liquid is concentrated under reduced pressure into the medical fluid that concentration is 1 gram of every milliliter of crude drug, is centrifuged off insoluble matter；Concentrated medicament is taken, nothing is slowly added to
Water-ethanol, the ultimate density 80% stirring while adding for making ethyl alcohol in solution, 4 DEG C of alcohol precipitation 12h；Suction filtration obtains light brown thick more
Sugar, after Thick many candies remove isolating protein with SAVEGE method, alcohol precipitation, is then successively washed 3 times with dehydrated alcohol, acetone again, transfer
Into evaporating dish, vacuum drying obtains attached art polysaccharide；
2) preparation of atractylodes lactone: weighing Rhizoma Atractylodis Macrocephalae 1000g, is added 75% ethyl alcohol, 100 DEG C water-bath refluxing extraction 2 times,
3 hours every time, 8 times of amount solvents every time；Combined extract, is concentrated under reduced pressure into the medical fluid that concentration is 1 gram of every milliliter of crude drug, and centrifugation removes
Insoluble matter is removed, adds to D-101 macroporous resin column upper end, standing adsorption 1 hour or so；6 times of column volume pure water elutions, discard and wash
De- liquid, 5 times of 70% ethanol elutions of column volume collect eluent, are concentrated into small size, and vacuum drying obtains atractylodes lactone；
3) preparation of Radix Aconiti Lateralis Preparata alkaloid: weighing RADIX ACONITI LATERALIS PREPARATA medicinal material 1000g, and 75% ethyl alcohol, 100 DEG C of water-bath refluxing extractions are added
2 times, 3 hours every time, 8 times of amount solvents every time；Combined extract is concentrated under reduced pressure into the medical fluid that concentration is 1 gram of every milliliter of crude drug, from
The heart removes insoluble matter, adds to D-101 macroporous resin column upper end, and standing adsorption 1 hour or so；6 times of column volume pure water elutions, are abandoned
Eluent is removed, 5 times of 70% ethanol elutions of column volume collect eluent, are concentrated into small size, and vacuum drying obtains monkshood biology
Alkali.
Preferably, the optimization of the attached art polysaccharide, atractylodes lactone and Radix Aconiti Lateralis Preparata alkaloid three's proportion compatibility uses
Simple checker board design carries out component proportion experiment, and carries out multiple-objection optimization with response surface and craving function method, and foundation has
Imitate the relationship between component and its compatibility proportion mode and drug effect.
Simple checker board has been used to design in the present invention, the ratio of optimum combination, group are sought in 10 groups of combinations in total
It is few, credible result.Compared with prior art, the active principle of monkshood and Rhizoma Atractylodis Macrocephalae medicine centering of the present invention is rationally matched
5, so that it is well used in terms of preparing drugs for rheumatoid arthritis, it is new to be found from complicated Chinese medicine system
Component compatibility new idea and method is provided, can be used as two kind new medicine of Chinese medicine exploitation.
Detailed description of the invention
Fig. 1 is the FT-IR spectrogram of attached art polysaccharide component.
Fig. 2 is the HPLC figure of atractylenolide (A, 276nm) in atractylodes lactone component, I, III (B, 220nm).
Fig. 3 is that the HPLC of Radix Aconiti Lateralis Preparata alkaloid active principle schemes.
Fig. 4 is the response surface figure and contour map that monkshood Rhizoma Atractylodis Macrocephalae medicine optimizes different prescriptions to left foot swelling degree of the paw.
Fig. 5 is the response surface figure and contour map that monkshood Rhizoma Atractylodis Macrocephalae medicine optimizes different prescriptions to IL-1 β in serum.
Fig. 6 is the response surface figure and contour map that monkshood Rhizoma Atractylodis Macrocephalae medicine optimizes different prescriptions to 5-HT in serum.
Fig. 7 is the response of D value binomial model and the distribution map of predicted value.
Specific embodiment
The present invention substantive content that present invention be described in more detail in conjunction with the accompanying drawings and embodiments, the embodiment are only used for
The bright present invention and not limitation of the present invention.
The preparation of one active principle
1. the preparation of the attached art polysaccharide of active principle
Monkshood and Rhizoma Atractylodis Macrocephalae 1000g (monkshood: Rhizoma Atractylodis Macrocephalae=3:1) are weighed, 6 times of amount volume petroleum ether (boiling points: 60- are added
90 DEG C) extract 2 times, after extracting 3 hours every time, collect the dregs of a decoction and be added 80% ethyl alcohol, 100 DEG C water-bath refluxing extraction 2 times, every time 3
Hour, 8 times of amount solvents every time, to remove the fat in medicinal material, pigment, monosaccharide and oligosaccharides.Collect 60 DEG C of vacuum drying 12h of the dregs of a decoction
Afterwards, add 100 DEG C of water of 10 times of amounts to extract 3 hours, extract altogether twice, combined extract, being concentrated under reduced pressure into concentration is 1 gram of every milli of crude drug
The medical fluid risen, is centrifuged off insoluble matter.Concentrated medicament is taken, dehydrated alcohol is slowly added to, it is stirring while adding to make ethyl alcohol in solution
Ultimate density be 80%, 4 DEG C of alcohol precipitation 12h.Suction filtration obtains light brown Thick many candies, after SAVEGE method removes isolating protein, alcohol again
Heavy, dehydrated alcohol, acetone successively wash 3 times, are transferred in evaporating dish, are dried in vacuo spare.
2. the preparation of active principle atractylodes lactone
Weigh Rhizoma Atractylodis Macrocephalae 1000g, be added 75% ethyl alcohol, 100 DEG C water-bath refluxing extraction 2 times, every time 3 hours, every time 8 times
Measure solvent.Combined extract is concentrated under reduced pressure into the medical fluid that concentration is 1 gram of every milliliter of crude drug, is centrifuged off insoluble matter, adds to D-
101 macroporous resin column upper ends, standing adsorption 1 hour or so.6 times of column volume pure water elutions, discard eluent, 5 times of column volumes
70% ethanol elution collects eluent, is concentrated into small size, is dried in vacuo spare.
3. the preparation of active principle Radix Aconiti Lateralis Preparata alkaloid
Weigh RADIX ACONITI LATERALIS PREPARATA medicinal material 1000g, be added 75% ethyl alcohol, 100 DEG C water-bath refluxing extraction 2 times, 3 hours every time, every time 8
Amount solvent again.Combined extract is concentrated under reduced pressure into the medical fluid that concentration is 1 gram of every milliliter of crude drug, is centrifuged off insoluble matter, adds to
D-101 macroporous resin column upper end, standing adsorption 1 hour or so.6 times of column volume pure water elutions, discard eluent, 5 times of column volumes
70% ethanol elution collects eluent, is concentrated into small size, is dried in vacuo spare.
The analysis of two active principles
1. attached art polysaccharide active principle analysis
Precision is weighed through 50 DEG C of vacuum drying polysaccharide 5mg, is dissolved in water, is settled in 50mL volumetric flask.Match
Solution is 0.1gmL-1Test solution.0.1mgmL is pipetted respectively-1Dextrose standard sample solution 0.2,0.4,0.6,
0.8,1.0,1.2mL adds distilled water to be supplemented to 2mL, using 2mL distilled water as blank control, by benzene in dry tool plug test tube
Phenol-sulfuric acid method measures absorbance A value at 489nm.And the polysaccharide sample solution 1mL prepared is taken to be placed in dry tool plug test tube,
Sample absorbance value is measured according to the above method.It is 65.16% according to the content that standard curve and sample concentration calculate wherein total reducing sugar.
Appropriate attached art polysaccharide sample is separately taken, using KBr pressed disc method, in 4000-400cm-1Infrared spectroscopy is carried out in range to sweep
It retouches, scanning times are 32 times, resolution ratio 4cm-1, the result is shown in Figure 1.By in figure it is found that polysaccharide sample is in 2929.42cm-1Place
The stretching vibration that absorption peak is C-H absorbs, 1418.94cm-1The bending vibration that the absorption peak at place is C-H absorbs, this two groups of absorptions
Peak is the characteristic absorption peak of saccharide compound；3421.95cm-1The strong absworption peak at place is that the stretching vibration of O-H key absorbs；
1644.93cm-1The absorption peak at place is the absorption in conjunction with water；1155.78,1080.65 and 1022.42cm-1The absorption peak at place is pyrrole
That mutters C=O in ring structure washes receipts peak, and pyranoside has 3 absorption peaks at 1100-1010；844.05cm-1The absorption peak at place is
The angle of α-D- pyranose C-H vibrates, and shows that the polysaccharide component contains α-D- pyranose glucose ring, molecule is connected with α-glycosidic bond
It connects.
2. atractylodes lactone active principle is analyzed
Homemade atractylodes lactone component 0.2632g is separately taken, methanol dissolves, and after ultrasonic 30min, centrifugation takes supernatant constant volume
To 10mL, 0.22 μm of organic film is crossed, it is spare to obtain test solution.
HPLC condition: chromatographic column C18(4.6mm × 250mm, 5 μm)；Mobile phase A: acetonitrile, Mobile phase B: water, gradient are washed
De- program is shown in Table 1；Flow velocity: 1mLmin-1；Column temperature: 30 DEG C；Sample volume: 10 μ L.
The condition of gradient elution of 1 atractylodes lactone component of table
| Sequence | Time (min) | A% | B% |
| 1 | 0 | 60 | 40 |
| 2 | 16 | 70 | 30 |
| 3 | 18 | 65 | 35 |
| 6 | 30 | 65 | 35 |
The efficient liquid phase chromatographic analysis spectrogram of atractylodes lactone active principle is shown in Fig. 2.HPLC measures white in atractylodes lactone component
The content of art lactone I, II, III is respectively 202.14mgg-1, 86.40mgg-1, 321.30mgg-1。
3. Radix Aconiti Lateralis Preparata alkaloid active principle is analyzed
Self-control alkaloidal constituent powder 0.1000g is weighed, ethyl alcohol 5ml is added to dissolve, adjusts PH to 10-11, chloroform extracts 2 times,
Each 10ml, takes chloroform layer to volatilize, and residue adds ethyl alcohol 5mL to make to dissolve, and sulfuric acid titrating solution (0.01molL is added in precision-1)
15mL, water 15mL and methyl red indicator solution 3 drip, with sodium hydroxide titration liquid (0.02molL-1) it is titrated to yellow.With 1mL sulphur
Acidometric titration liquid (0.01molL-1) it is equivalent to the rhizome of Chinese monkshood (C of 12.9mg34H47NO11) calculate, total biology in 0.1g alkaloidal constituent
The content of alkali is 52.096mg.
Precision weighs homemade alkaloid active principle 0.1008g, is dissolved in 10mL methanol, ultrasonic 30min, and centrifugation takes
Clear liquid adds methanol constant volume to 10mL, crosses 0.22 μm of organic film, it is spare to obtain test solution.
HPLC condition: chromatographic column is Dalian Yi Lite BDS C18Column (4.6 × 250mm, 5 μm)；Mobile phase: methanol:
0.15% triethylamine-aqueous solution gradient elution, gradient condition are shown in Table 2, flow velocity 1mLmin-1。
The elution requirement of 2 alkaloidal constituent of table
| Time | Methanol | 0.15% triethylamine-aqueous solution |
| 0 | 55 | 45 |
| 7 | 56 | 44 |
| 10 | 57 | 43 |
| 15 | 60 | 40 |
| 30 | 69 | 31 |
| 35 | 73 | 27 |
| 40 | 75 | 25 |
The efficient liquid phase chromatographic analysis spectrogram of Radix Aconiti Lateralis Preparata alkaloid active principle is shown in that Fig. 3, HPLC measure crow in alkaloidal constituent
Head alkali, mesaconine and Hypaconitine content are respectively 72.10mgg-1, 96.24mgg-1, 367.76mgg-1。
The effect experiment of three different active principle compatibility proportions
This experiment uses simple checker board experimental design, selects 3 kinds of components of monkshood Rhizoma Atractylodis Macrocephalae medicine centering, monkshood soluble polysaccharide
(A), atractylodes lactone (B) and Radix Aconiti Lateralis Preparata total alkaloids (C) constitute 3 factors, (300mgkgd under same dosage-1),
Using { 3,3 } simplex lattice design, testing site number is C3 3 +3-1=10.Calculated to simplify, the concentration of each component into
Standardization is gone, Cmin is classified as 0, and maximum concentration is classified as 1.Each point Xi value is 0,1/3,2/3 and 1.Planning of experiments such as table 3
It is shown.Your wherein had because containing toxic stronger diester-type alkaloids according to diester-type alkaloids in Radix Aconiti Lateralis Preparata total alkaloids component
The maximum dose of the regulation of limitation, diester-type alkaloids involved in text is below the LD of rat in preliminary experiment50。
The simple checker board experimental design table of table 3
| Group | Active principle A | Active principle B | Active principle C | Drug effect Y |
| C1 | 1 | 0 | 0 | Y1 |
| C2 | 0 | 1 | 0 | Y2 |
| C3 | 0 | 0 | 1 | Y3 |
| C4 | 2/3 | 1/3 | 0 | Y4 |
| C5 | 2/3 | 0 | 1/3 | Y5 |
| C6 | 1/3 | 0 | 2/3 | Y6 |
| C7 | 1/3 | 2/3 | 0 | Y7 |
| C8 | 0 | 1/3 | 2/3 | Y8 |
| C9 | 0 | 2/3 | 1/3 | Y9 |
| C10 | 1/3 | 1/3 | 1/3 | Y10 |
After right hind foot of rat routine disinfection, Freund's complete adjuvant 0.02mL system is injected to vola pedis pad portion to ankle-joint direction
Make adjuvant-induced arthritis animal model.Modeling success SD rat is selected, second day after modeling starts to be administered, according to simple grid
Three components are combined into 10 prescriptions by shape design, are divided into 10 groups, every group of 8 rats, daily 300mgkg-1Mouse stomach-filling again
In addition administration arranges model group and normal group to give isometric physiological saline.Successive administration put to death animal after 21 days, took blood
3mL is set in Eppendorf pipe, and 3500rpm is centrifuged 15min, takes serum spare, and ELISA method measures IL-1 β content in serum；Together
When remove full brain, take 1/2 fresh brain tissue, after being cleaned, dried with physiological saline, being weighed, homogenate, 3000rpm be centrifuged 15min,
Supernatant is taken, ELISA method measures the content of 5-HT in maincenter brain tissue.Left foot vola pedis thickness is measured with thickness gauge and calculates vola pedis
Swelling.
Table 4 be after monkshood Rhizoma Atractylodis Macrocephalae and the administration of different prescription to adjuvant arthritis rats left foot swelling degree of the paw, IL-1 β with
And the influence of 5-HT.It can be seen from table after rat modeling, swelling degree of the paw and IL-1 β content, which have, largely to be risen
High (P<0.05, P<0.05), and cental system 5-HT content because after modeling pain stimulation be increased slightly (P>0.05).Attached art is more
Sugar, atractylodes lactone and Radix Aconiti Lateralis Preparata alkaloid single component have extremely significant influence to swelling degree of the paw, IL-1 β and 5-HT, and P value is equal
Less than 0.01.And the content of IL-1 β, rises in the swelling degree of the paw and serum of the extremely significant reduction rat of all prescriptions energy
The content of 5-HT in high brain tissue, P value are respectively less than 0.01.
4 monkshood Rhizoma Atractylodis Macrocephalae difference prescription of table to swelling degree of the paw, IL-1 β and 5-HT influence (N=8)
Note: compared with blank group▲P < 0.05,▲▲P<0.01；Compared with model group*P < 0.05,**P<0.01。
The optimization of four active principle compatibilities proportion
Although having there is many methods as evolved and genetic algorithm is for multiple-objection optimization at present, in view of response surface and
Crave for unique advantage of the function in optimization, therefore completes compatibility optimization using Response Surface Method and craving function.Wherein respond
Face method is carried out using Design Expert 8.0.6 software, reflects the pass between variable and response with binary polynomial equation
System；And crave for function can be by the way that multiple response variables be converted to craving value to reaching desired optimal value, herein, I
First by each index value YiValue is converted to independent craving value d respectively, and craving value is between 0~1, if response exists
When except acceptable range, if di=0, if its response is considered as perfectly, if di=1.What if response was pursued
It is to be the bigger the better, then specific calculation formula is as follows:
If pursue response be it is the smaller the better, specific calculation formula is as follows:
Wherein Yi-minFor acceptable minimum YiIndex value, and Yi-maxIt is then the largest index value, r is normal number.Finally
Whole craving function D value is calculated in conjunction with all independent craving values, then by following formula to realize total optimization:
Wherein D indicates whole craving value, diIndicate independent craving value, WiIndicate the weight coefficient of i-th of d value.
According to variance analysis, the multivariate regression equation of swelling degree of the paw, IL-1 β and 5-HT is obtained, use (1), (2) respectively
(3) it indicates.
Y1=31.05A+28.78B+31.16C-7.15AB-10.59AC-24.99BC (1)
Y2=53.39A+53.35B+47.46C-36.28AB-38.34AC-30.29BC (2)
Y3=662.38A+645.44B+708.42C+924.58AB+1162.29AC+1448.59BC (3)
It can be seen that three components share and pedal swelling can be significantly reduced from the contour and response surface figure of Fig. 4-Fig. 6
The content of IL-1 β in degree and serum increases the content of 5-HT in brain tissue.Wherein, atractylodes lactone component and Radix Aconiti Lateralis Preparata alkaloid
The effect of compatibility is than more significant.Three YiIt is worth (Y1, Y2, Y3) it is converted to corresponding d respectivelyiIt is worth (d1, d2, d3), in order to reach monkshood
Three optimal anti-inflammatory pain-stopping effects of component of Rhizoma Atractylodis Macrocephalae medicine centering, rat left foot swollen tissue degree (d1) value it is the smaller the better, blood
Clear IL-1 β (d2) value be the bigger the better, the value (d of 5-HT in brain tissue3) be the bigger the better.Therefore, by rat left foot swollen tissue degree
(d1) minimum value be set as 1.08%, maximum value is then set as 100%；- 1 β (d of serum IL2) theoretical minimum value be 0, maximum value is then
It is set as 100pgml-1；5-HT (d in brain tissue3) content theoretical minimum value be 0, maximum value is set as 1500ngml-1.Cause
This, the value of d1, d2, d3 are respectively calculated as follows:
Therefore, the calculating of whole craving value D is as follows:
D=(d1d2d3)1/3
The independent craving value d of final resulting each index1、d2、d3And whole the specific of craving value D is shown in Table 5, with response
Surface Method optimizes D value, obtains regression equation are as follows:
D=0.52A+0.52B+0.56C+0.39AB+0.45AC+0.54BC+0.16ABC
The craving functional value of 5 monkshood Rhizoma Atractylodis Macrocephalae difference prescription of table
Fig. 7 is the distribution of the response and predicted value of D value binomial model, be can be seen from the chart, most point is all
It falls near diagonal line, the deviation between response and predicted value is little, shows that models fitting data are good.It is excellent using response surface
Change, the optimum combination finally filtered out is that monkshood soluble polysaccharide (A), atractylodes lactone (B) and Radix Aconiti Lateralis Preparata alkaloid (C) account for respectively
25.5%, 33.0% and 41.5%, i.e. 51:66:83.
The verifying of five optimization prescriptions
In order to verify calculated result, experimental verification has been carried out with identical animal model.The best compatibility that optimization is obtained
Side respectively compares compatibility group with atractylodes lactone+Radix Aconiti Lateralis Preparata alkaloid (V1 group, 1:1), blank group, model group, monkshood Rhizoma Atractylodis Macrocephalae medicine
Compared with research.As shown in table 6, compared with model group, best compatibility can be substantially reduced -1 β content (P of swelling degree of the paw and serum IL
< 0.01, P < 0.01), and can increase the content (P < 0.01) of 5-HT in brain；Simultaneously best compatibility side reduce swelling degree of the paw and
The effect of 5-HT content is compared with monkshood Rhizoma Atractylodis Macrocephalae medicine is to compatibility group in -1 β content of serum IL and raising brain, in addition to swelling degree of the paw
Outside without significant statistical significance (P < 0.05), i.e., without significant difference outside, other two index is superior to monkshood Rhizoma Atractylodis Macrocephalae medicine to matching
5 groups, have significant statistical significance (P < 0.05, P < 0.05)；The anti-inflammatory pain-stopping effect of i.e. best compatibility side is white better than monkshood
Art medicine is to compatibility.
6 monkshood Rhizoma Atractylodis Macrocephalae medicine of table to component compatibility optimization side to swelling degree of the paw, IL-1 β and 5-HT influence (N=
8)
Note: compared with blank group▲P < 0.05,▲▲P<0.01；Compared with model group*P < 0.05,**P<0.01；It is white with monkshood
Art medicine compares compatibility group#P<0.05,##P<0.01。
Simple checker board has been used to design in the present invention, the ratio of optimum combination, group are sought in 10 groups of combinations in total
It is few, credible result.Compared with prior art, the active principle of monkshood and Rhizoma Atractylodis Macrocephalae medicine centering of the present invention is rationally matched
5, so that it is well used in terms of preparing drugs for rheumatoid arthritis, it is new to be found from complicated Chinese medicine system
Component compatibility new idea and method is provided, can be used as two kind new medicine of Chinese medicine exploitation.
Claims (4)
1. a kind of Chinese medicine composition for treating rheumatoid arthritis, which is characterized in that be made of the extract of following parts by weight: attached
Art polysaccharide 15~35%, atractylodes lactone 23~43%, Radix Aconiti Lateralis Preparata alkaloid 30~50%,
1) the attached art polysaccharide the preparation method is as follows: weigh monkshood and Rhizoma Atractylodis Macrocephalae 1000g, wherein monkshood dosage is Rhizoma Atractylodis Macrocephalae
3 times, 6 times of amount volumes and boiling point are added and are extracted 2 times for 60 DEG C -90 DEG C of petroleum ether, after extracting 3 hours every time, collects the dregs of a decoction and add
Enter 80% ethyl alcohol, 100 DEG C water-bath refluxing extraction 2 times, 3 hours every time, 8 times of amount solvents, removed fat, the color in medicinal material every time
Element, monosaccharide and oligosaccharides；After collecting 60 DEG C of vacuum drying 12h of the dregs of a decoction, add 100 DEG C of water of 10 times of amounts to extract 3 hours, extract altogether twice,
Combined extract is concentrated under reduced pressure into the medical fluid that concentration is 1 gram of every milliliter of crude drug, is centrifuged off insoluble matter；Concentrated medicament is taken, is delayed
It is slow that dehydrated alcohol, the ultimate density 80% stirring while adding for making ethyl alcohol in solution, 4 DEG C of alcohol precipitation 12h are added；Suction filtration obtains shallow brown
Color Thick many candies, after Thick many candies remove isolating protein with SAVEGE method, alcohol precipitation, then successively washs 3 with dehydrated alcohol, acetone again
It is secondary, it is transferred in evaporating dish, vacuum drying obtains attached art polysaccharide；
2) atractylodes lactone the preparation method is as follows: weigh Rhizoma Atractylodis Macrocephalae 1000g, 75% ethyl alcohol, 100 DEG C of water-baths reflux are added
Extract 2 times, 3 hours every time, 8 times of amount solvents every time；Combined extract is concentrated under reduced pressure into the medicine that concentration is 1 gram of every milliliter of crude drug
Liquid is centrifuged off insoluble matter, adds to D-101 macroporous resin column upper end, and standing adsorption 1 hour；6 times of column volume pure water elutions,
Eluent is discarded, 5 times of 70% ethanol elutions of column volume collect eluent, are concentrated into small size, and vacuum drying obtains in Rhizoma Atractylodis Macrocephalae
Ester；
3) Radix Aconiti Lateralis Preparata alkaloid the preparation method is as follows: weigh RADIX ACONITI LATERALIS PREPARATA medicinal material 1000g, 75% ethyl alcohol, 100 DEG C of water-baths are added
Refluxing extraction 2 times, 3 hours every time, 8 times of amount solvents every time；Combined extract, being concentrated under reduced pressure into concentration is 1 gram of every milliliter of crude drug
Medical fluid, be centrifuged off insoluble matter, add to D-101 macroporous resin column upper end, standing adsorption 1 hour；6 times of column volume pure water are washed
It is de-, eluent is discarded, 5 times of 70% ethanol elutions of column volume collect eluent, are concentrated into small size, and vacuum drying obtains monkshood
Alkaloid.
2. a kind of Chinese medicine composition for treating rheumatoid arthritis according to claim 1, which is characterized in that the attached art
The ratio of polysaccharide is 25.5%, and the ratio of atractylodes lactone is 33.0%, and the ratio of Radix Aconiti Lateralis Preparata alkaloid is 41.5%.
3. a kind of Chinese medicine composition for treating rheumatoid arthritis according to claim 1 or 2, which is characterized in that described
Attached art polysaccharide is extracted by monkshood and Rhizoma Atractylodis Macrocephalae, and the atractylodes lactone is extracted by Rhizoma Atractylodis Macrocephalae, and the Radix Aconiti Lateralis Preparata alkaloid is extracted by RADIX ACONITI LATERALIS PREPARATA.
4. a kind of Chinese medicine composition for treating rheumatoid arthritis according to claim 1, which is characterized in that the attached art
The optimization of polysaccharide, atractylodes lactone and Radix Aconiti Lateralis Preparata alkaloid three's proportion compatibility is designed using simple checker board and carries out component proportion reality
It tests, and carries out multiple-objection optimization with response surface and craving function method, establish active principle and its compatibility proportion mode and drug effect
Between relationship.
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