CN109157562A - Anti-ulcerative colitis granule and preparation method - Google Patents
Anti-ulcerative colitis granule and preparation method Download PDFInfo
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Abstract
The invention belongs to field of medicaments, being related to a kind of anti-ulcerative colitis granule and preparation method, anti-ulcerative colitis granule is prepared by purslane and cortex cinnamomi；The starting weight of purslane and cortex cinnamomi ratio is 10-14:3-7.A kind of that the present invention provides preparation processes is simple, has no toxic side effect and can be used as treatment ulcerative colitis anti-ulcerative colitis granule and preparation method and granule.
Description
Technical field
The invention belongs to field of medicaments, it is related to a kind of granule and preparation method more particularly to a kind of anti-ulcerative colon
Scorching granule and preparation method.
Background technique
Ulcerative colitis (Ulcerative colitis, UC) is that a kind of nonspecific diseases associated with inflammation is related to mucous membrane
With colorectal mucosa lower layer rectum.Under special circumstances, it is defined as colitis, entire large intestine may be will affect.Clinical manifestation is main
For abdominal pain, diarrhea, mucus pus and blood stool etc., colon is glutinous in diffusivities inflammation, Histopathologic appearances such as erosion, ulcer under scope
Diffusivity inflammatory reaction, goblet cell reduction, crypts structure are destroyed under film or mucous membrane.Research report display UC generation and in vivo
Press down the scorching factor and urges the unbalance closely related of the scorching factor.Inflammatory cytokine abnormal increase or suppression inflammatory cytokines reduce equal extremely
It can promote the variation of intestinal mucosa inflammation, and lasting immunological regulation is unbalance, and inflammation is promoted to tend to chronicity.Ulcerative colitis
Active stage IL-10 expression reduces, and TNF-α expression increases, and makes vivo immunization Imbalance, thus induced ulcer knot
Enteritis.China UC morbidity at present is in rising trend.Correlation seminar of Shaanxi university of TCM finds that purslane is more in early-stage study
Sugar has good therapeutic effect to the UC mouse model that DSS is induced, and can obviously improve the pathologic structure of mouse Colon, balanced body
Interior inflammatory factor and the level for pressing down the scorching factor.It has been reported that cortex cinnamomi Aqueous extracts have good curative effect to UC.In the prior art simultaneously
It does not directly adopt purslane and cortex cinnamomi forms the drug or pharmaceutical composition of anti-ulcerative colitis.
Summary of the invention
In order to solve the above technical problems in background technology, simple, nothing that the present invention provides a kind of preparation processes
Toxic side effect and the anti-ulcerative colitis granule and preparation method that can be used as treatment ulcerative colitis.
To achieve the goals above, the present invention adopts the following technical scheme:
A kind of anti-ulcerative colitis granule, it is characterised in that: the anti-ulcerative colitis granule is by dent
Amaranth and cortex cinnamomi are prepared；The starting weight of the purslane and cortex cinnamomi ratio is 10-14:3-7.
The weight ratio of above-mentioned purslane and cortex cinnamomi is 11-12:4-5.
A kind of preparation method of anti-ulcerative colitis granule, it is characterised in that: the anti-ulcerative colitis particle
The preparation method of agent the following steps are included:
1) it is that the ratio of 10-14:3-7 weighs purslane and cortex cinnamomi and mixes purslane with cortex cinnamomi by weight, obtains
To mixture；
2) water is added in the obtained mixture of step 1), 60 DEG C -100 DEG C at a temperature of extract 1-6h, filter medicine
Slag is concentrated into thick medicinal extract；
3) thick medicinal extract that step 2) is prepared was placed in -80 DEG C of refrigerator freezings not less than 24 hours, and taking-up is placed on cold
Freeze drying in drier, is ground into fine powder after the completion；
4) fine powder for taking step 3) to be prepared is mixed with sucrose；The weight ratio of the fine powder and sucrose is 1:2-5；
5) 80% ethanol solution is added into the mixture that step 4) is prepared, hand is made and pinches what agglomerating, flicking dissipated
Softwood；
6) particle is made by No. 1 medicine sieve in a manner of squeezing granulation in the softwood that step 5) is prepared.
Above-mentioned steps 1) in, the weight ratio of purslane and cortex cinnamomi is 11-12:4-5.
Above-mentioned steps 2) in the solid-liquid ratio of mixture and water be 1:10-50.
Above-mentioned steps 2) in the solid-liquid ratio of mixture and water be 1:40.
Above-mentioned steps 2) Extracting temperature be 90 DEG C, extraction time is 5h.
Above-mentioned steps 3) in dry time be for 24 hours.
Above-mentioned steps 4) in, the weight ratio of fine powder and sucrose is not less than 1:3.
The preparation method of above-mentioned anti-ulcerative colitis granule is after step 6) further include:
7) particle that step 6) is prepared is put into baking oven dry, with No. 1 screens out big particle after dry, and No. 4
Fine powder is screened out, finished product is obtained.
The invention has the advantages that
The present invention provides a kind of anti-ulcerative colitis granule and preparation method, the anti-ulcerative colitis granules
It is prepared by purslane and cortex cinnamomi；The starting weight of purslane and cortex cinnamomi ratio is 10-14:3-7.The preparation method is 1) to press
Weight ratio is that the ratio of 10-14:3-7 weighs purslane and cortex cinnamomi and mixes purslane with cortex cinnamomi, obtains mixture；2) exist
Water is added in the obtained mixture of step 1), 60 DEG C -100 DEG C at a temperature of extract 1-6h, filter the dregs of a decoction, be concentrated into thick leaching
Cream；3) thick medicinal extract that step 2) is prepared was placed in -80 DEG C of refrigerator freezings not less than 24 hours, and taking-up is placed on freeze-drying
It is dry in device, it is ground into fine powder after the completion；4) fine powder for taking step 3) to be prepared is mixed with sucrose；The weight of fine powder and sucrose
Than being 1:2-5；5) 80% ethanol solution is added into the mixture that step 4) is prepared, be made hand pinch agglomerating, flicking i.e. dissipate
Softwood；6) particle is made by No. 1 medicine sieve in a manner of squeezing granulation in the softwood that step 5) is prepared.The present invention selects
Fever and chills medicine is to purslane and cortex cinnamomi compatibility, and wherein purslane is sour cold in nature, returns large intestine, Liver Channel, have it is clearing heat and detoxicating, remove heat from the blood and relieve diarrhea
The effect of, it is usually used in treating damp-heat diarrhea, metrorrhagia and metrostaxis hematochezia is clinical treatment ulcerative colitis conventional Chinese medicine；Cortex cinnamomi is common
Drugs for dispelling internal cold, acrid flavour heat, returns spleen, kidney, the heart, Liver Channel have the effect of mends fire supporing yang, eliminating cold to stop pain, warming meridians and promoting circulation of qi；Cortex cinnamomi contains osmanthus
The ingredients such as skin aldehyde have abirritant effect to gastrointestinal mucosa, and can promote long movement, increase digestive secretion, enhancing
Digestive function excludes alimentary canal pneumatosis, alleviates pain caused by gastrointestinal convulsion.Purslane and cortex cinnamomi compatibility, the cold heat of the two one,
One monarch, one minister, purslane cure mainly the chief complaint of ulcerative colitis, and cortex cinnamomi is warming channel and expelling cold, alleviate the cold of purslane.And two
Person is integration of drinking and medicinal herbs Chinese medicine, is had no toxic side effect, and can be used as the drug for the treatment of ulcerative colitis, is taken for a long time.
Detailed description of the invention
Fig. 1 is the influence diagram of DSS induction UC mouse Signs；
Fig. 2 is the H&E coloration result such as following figure of different experiments group；
Fig. 3 is the Wei Entu of different experiments group OUT number analysis；
Fig. 4 is the variation diagram of different sample interior intestinal flora bio-diversities；
Fig. 5 is the result figure of the granule readjustment flora of purslane and cortex cinnamomi.
Specific embodiment
The present invention provides a kind of anti-ulcerative colitis granules, are prepared by purslane and cortex cinnamomi；Purslane
Starting weight ratio with cortex cinnamomi is 10-14:3-7, and preferred proportion is 11-12:4-5.
At the same time, the present invention also provides a kind of preparation method of anti-ulcerative colitis granule, this method includes
Following steps:
1) it is that the ratio of 10-14:3-7 weighs purslane and cortex cinnamomi and mixes purslane with cortex cinnamomi by weight, obtains
To mixture, the preferred 11-12:4-5 of the weight ratio of purslane and cortex cinnamomi.2) water is added in the obtained mixture of step 1),
60 DEG C -100 DEG C (preferably 90 DEG C) at a temperature of extract 1-6h (preferably 5h), filter the dregs of a decoction, be concentrated into thick medicinal extract；Mixture with
The solid-liquid ratio of water is 1:10-50, preferably 1:40；
3) thick medicinal extract that step 2) is prepared was placed in -80 DEG C of refrigerator freezings not less than 24 hours, and taking-up is placed on cold
Freeze drying in drier and for 24 hours, is ground into fine powder after the completion；
4) fine powder for taking step 3) to be prepared is mixed with sucrose；The weight ratio of the fine powder and sucrose is 1:2-5, preferably
Not less than 1:3；
5) 80% ethanol solution is added into the mixture that step 4) is prepared, hand is made and pinches what agglomerating, flicking dissipated
Softwood；
6) particle is made by No. 1 medicine sieve in a manner of squeezing granulation in the softwood that step 5) is prepared.
7) particle that step 6) is prepared is put into baking oven dry, with No. 1 screens out big particle after dry, and No. 4
Fine powder is screened out, finished product is obtained.
Theoretical basis of the invention is:
Fever and chills medicine selected by the present invention is Shaanxi State Administration of Traditional Chinese Medicine of university of TCM taste to purslane and cortex cinnamomi
Ulcerative colitis herbal pair is treated in sick academic leader Liu Li professor clinic, proves its treatment by decades clinical practice
Effect is definite.And (class is studied to the mechanism of action of purslane under the support that project is cultivated in the industrialization of Shaanxi Department of Education of Shanxi Province
Autograph claims: the research and development of Portulaca oleracea linn chewable tablet, number: 2011JG19).Fever and chills medicine is sour to purslane in compatibility cold in nature,
Return large intestine, Liver Channel, have it is clearing heat and detoxicating, the effect of removing heat from the blood and relieve diarrhea, be usually used in treat damp-heat diarrhea, metrorrhagia and metrostaxis hematochezia is that clinic is controlled
Treat ulcerative colitis conventional Chinese medicine.Cortex cinnamomi is common drugs for dispelling internal cold, and acrid flavour heat, there is benefit fire to help for returns spleen, kidney, the heart, Liver Channel
The effect of sun, eliminating cold to stop pain, warming meridians and promoting circulation of qi.Cortex cinnamomi has abirritant effect containing ingredients such as cinnaldehydrum, to gastrointestinal mucosa, and
It can promote long movement, increase digestive secretion, enhancing digestive function, exclude alimentary canal pneumatosis, alleviate caused by gastrointestinal convulsion
Pain.Purslane and cortex cinnamomi compatibility, the cold heat of the two one, one minister of a monarch, purslane cure mainly the chief complaint of ulcerative colitis,
Cortex cinnamomi is warming channel and expelling cold, alleviates the cold of purslane.And the two is all integration of drinking and medicinal herbs Chinese medicine, is had no toxic side effect, and can be used as treatment
The drug of ulcerative colitis is taken for a long time.
1 instrument and material
1.1 laboratory apparatus
Electric-heated thermostatic water bath (Yongxing Ke Wei, Beijing Instrument Ltd.)；Ultraviolet-visible spectrophotometer UV-2006
(Shimadzu Co., Ltd)；FA2004B type electronic balance (Shanghai You Ke instrument and meter Co., Ltd)；101 type electric heating forced air dryings
Case (Yongxing Ke Wei, Beijing Instrument Ltd.)；IMS-50 ice machine (Xue Ke Electrical Appliances Co., Ltd of Changshu City)；Mouse ELISA examination
Agent box (Xin Bosheng Biotechnology Co., Ltd) microplate reader (Multiskan GO) -1510 (Thermo Fisher).
1.2 experimental material
Purslane Chinese medicine (Shaanxi prosperity moral pharmaceutcal corporation, Ltd, lot number: 20170401)；(Shaanxi is emerging for cortex cinnamomi Chinese medicine
Sheng De pharmaceutcal corporation, Ltd, lot number: 20170301)；Sugared (the lot number: S08J6G1 mass fraction >=98%, Shanghai of the anhydrous glucose of D-
Yuan Ye Biotechnology Co., Ltd)；The concentrated sulfuric acid (Xi'an San Pu chemical reagent Co., Ltd)；Phenol (Tianjin day power chemistry examination
Agent Co., Ltd).The extraction powder of purslane compatibility cortex cinnamomi；Dextran sulfate sodium MW:36000-50000 (DSS, MP
Biomedicals Biomedicines, Inc.)；Sulfasalazine (the good magnificent medicine company in Shanghai Xinyi).
2 methods and result
The research of 2.1 purslanes-cortex cinnamomi combination extraction process
Early-stage study finds that Polysaccharide from Portulaca oleracea has exact therapeutic effect to UC mouse, therefore purslane-cortex cinnamomi combination is most
Good proportion evaluation uses Polysaccharide from Portulaca oleracea content for index, and with the common proportion purslane of clinic: cortex cinnamomi (12:5) is reference, this hair
It is bright with purslane: cortex cinnamomi (12:5) is core, is extended and is extended accordingly outward, through inventor's long-term practice, purslane:
Result and purslane of the cortex cinnamomi in 10-14:3-7: the result of cortex cinnamomi (12:5) tends to be identical, especially purslane: cortex cinnamomi is in 11-
12:4-5 more shows very strong homoplasy.
Below by purslane: for cortex cinnamomi (12:5), being carried out such as to experimental condition of the invention and respective reaction parameter
Lower design:
2.1.1 the drafting of polysaccharide standard curve
Standard curve is drawn with phend-sulphuric acid, obtaining curve and obtaining regression equation is y=5.4616x+0.0923 (R2=
0.9985), the range of linearity is 0.025~0.15mg/mL.
2.1.2 the calculating of polysaccharide extract rate
Polysaccharide concentration is sought according to standard curve, calculates the content of polysaccharide in Chinese medicine.Polysaccharide extract rate is that polysaccharide accounts for dent
The percentage of amaranth compatibility cortex cinnamomi crude drug quality.
2.1.3 single factor experiment
2.1.3.1 influence of the ratio of purslane and cortex cinnamomi to polysaccharide extract rate:
It the results are shown in Table 1, it can be deduced that purslane and when cortex cinnamomi 12:5, recovery rate is higher.
The influence with comparison polysaccharide extract rate of 1 purslane of table and cortex cinnamomi
2.1.3.2 influence of the solid-liquid ratio to polysaccharide extract rate
It the results are shown in Table 2, it can be deduced that recovery rate highest when solid-liquid ratio 1:40.
Influence of 2 solid-liquid ratio of table to polysaccharide extract rate
2.1.3.3 influence of the Extracting temperature to polysaccharide extract rate
It the results are shown in Table 3, it can be deduced that as the temperature rises, recovery rate is also rising.
Influence of 3 Extracting temperature of table to polysaccharide extract rate
2.1.3.4 influence of the extraction time to polysaccharide extract rate
It the results are shown in Table 4, it can be deduced that when extraction time 1h and 2h, 4h and 5h, recovery rate increasing degree very little, 2h to 4h
When, with the extension of extraction time, recovery rate increases.
Influence of 4 extraction time of table to polysaccharide extract rate
2.1.4 orthogonal test
On the basis of single factor experiment, using polysaccharide extract rate as inspection target, with the horizontal L of four factor three9(43) carry out just
Test is handed over, referring to table 5 and table 6.
2.1.4.1 factor level table
5 factor level table of table
2.1.4.2 orthogonal test
6 orthogonal test of table
Orthogonal experiments show that optimum process is A3B1C3D2, i.e. purslane and cortex cinnamomi 12:5, solid-liquid ratio 1:30 extracts
Time 5h, 90 DEG C of Extracting temperature.It can be seen that influence of C (extraction time) factor to polysaccharide extract rate is maximum by R value.Four because
The size that element influences polysaccharide extract rate is as follows: C (extraction time) > B (solid-liquid ratio) > D (Extracting temperature) > A (proportion).
2.1.5 brief summary
Extraction process from four proportion of purslane and cortex cinnamomi, solid-liquid ratio, extraction time, Extracting temperature Elements research, with
Polysaccharide extract rate is that inspection target screens optimal experimental program.That is purslane and cortex cinnamomi 12:5, solid-liquid ratio 1:30, extraction time
5h, 90 DEG C of Extracting temperature are optimal extraction process.
3. the preparation of granule
3.1. experiment reagent and material
, cortex cinnamomi (Shaanxi prosperity moral Portulaca oleracea L (Shaanxi prosperity moral medicine company Co., Ltd, lot number: 20170401)
, dehydrated alcohol (Tianjin Tian Li chemical reagent Co., Ltd), pure water medicine company Co., Ltd, lot number: 20170301)
Deng.
2. laboratory apparatus
HH-S4 type digital display thermostat water bath, the Yongxing Ke Wei, Beijing Instrument Ltd., 101 type electric drying oven with forced convection north
The Yongxing Jing Kewei Instrument Ltd., VaCo type frozen drying instrument, Zirbus technology.
3. experimental method
The preparation of 3.1 purslanes-cortex cinnamomi granule
Embodiment 1:
Purslane 200.0g, cortex cinnamomi 83.3g are weighed, the water of 30 times of amounts is added, 90 DEG C of extraction 5h filter the dregs of a decoction, are concentrated into thick
Medicinal extract obtains 63.6g, is equivalent to crude drug 21.3g/100g.It is placed in -80 DEG C of refrigerator freezings for 24 hours, is subsequently placed in freeze-dryer
Middle drying for 24 hours, takes out, is ground into fine powder, obtains 43.1g cream powder, spare.It equally extracts again, is concentrated into thick paste, 4 times, saves thick leaching
Cream.
It weighs cream powder and sucrose to be uniformly mixed in 1:2 ratio, 80% ethanol solution is added and adjusts humidity, is made that " hand is pinched into
Group, flicking be dissipate " softwood.Particle is made by No. 1 medicine sieve in a manner of squeezing granulation again.It is put into drying in baking oven in time,
It can partially agglomerate after drying, adhesion, grind pressure.Big particle is screened out with No. 1, No. 4 screen out fine powder.To obtain the final product.
It weighs thick paste and sucrose to be uniformly mixed in 1:3,1:4,1:5 ratio, preparation method is same as above.
According to the embodiment 1 as above recorded, the present inventor passes through long-term practice, obtains matching embodiment as follows:
7 embodiment summary table of table
By taking embodiment 1 as an example, following detailed analysis and explanation are carried out to the granule that the present invention is prepared:
The quality examination of 3.2 purslanes-cortex cinnamomi particle
3.2.1 appearance character: particle should be dried, and particle is uniform, color is consistent, and the no moisture absorption, deliquesces phenomenon at agglomeration.
3.2.2 melting: weighing the particle 5.0g of the different auxiliary ratios of cream respectively, and hot water 100.0ml is added, and stirs 5 minutes, stands
Observe.
3.2.3 granularity: weighing the particle 30g of the different auxiliary ratios of cream respectively, the particle that cannot be sieved by No. 1 and can be by No. 5
The summation of the powder of sieve must not exceed 15%.
3.3 experimental result
3.3.1 granular mass: the auxiliary particle than 1:2,1:3,1:4,1:5 of cream be respectively 41.2g, 60.3g, 90.2g,
93.2g。
3.3.2 appearance character: the particle of variant cream auxiliary material ratio is dry, and particle is uniform, color is consistent, the no moisture absorption, knot
Block deliquesces phenomenon.
3.3.3 8 melting: be the results are shown in Table.
The melting of the particle of the auxiliary ratio of the different cream of table 8
3.3.4 granularity
The particle that cannot be sieved by No. 1 is respectively as follows: the auxiliary particle summation than 1:2 of cream with the powder summation that can be sieved by No. 5
Accounting is 8.1%, and the auxiliary particle summation accounting than 1:3 of cream is 10.2%, and the auxiliary particle summation accounting than 1:4 of cream is 9.8%, cream
The auxiliary particle summation accounting than 1:5 is 9.6%.
4. the measurement of the physical-chemical parameters of particle solution and dilution
4.1. laboratory apparatus
The laboratory PHSJ-4F PH meter, instrument electricity scientific instrument, DDS-307 conductivity meter, upper Nereid section, ZEN3600,
Malvern Malvern instrument
4.2. experimental method
4.2.1 the preparation of particle liquid
Precision weighs the particle 1.0g of the auxiliary ratio of each cream, the dissolution of 10ml distilled water is added, and dilute 10,50,100 times respectively,
It is spare.
4.2.2 the measurement of the physical-chemical parameters
Grain diameter: by the particle liquid of preparation, the measurement of grain diameter is carried out with Malvern instrument, records data.
Viscosity: by the particle liquid of preparation, the measurement of viscosity is carried out with viscosimeter, records data.
PH: by the particle liquid of preparation, the measurement of pH value is carried out with pH meter, records data.
4.3. experimental result and discussion
4.3.1 grain diameter (nm), experimental result is as shown in table 9, table 10, table 11 and table 12:
9 cream of table is auxiliary than 1:2 grain diameter
10 cream of table is auxiliary than 1:3 grain diameter
11 cream of table is auxiliary than 1:4 grain diameter
12 cream of table is auxiliary than 1:5 grain diameter
It is more undiluted small by the particle liquid partial size after the visible dilution of the above table, undiluted particle liquid dispersion effect compared with
Difference, particle can press together, so that partial size is larger.Cream it is auxiliary than 1:3 particle dilute 50,100 times after gap it is smaller, pdI value difference
It is worth also smaller, closest true partial size, and cream is auxiliary smaller than the grain diameter of 1:3.
4.3.2 viscosity (CP), as shown in table 13:
Each cream of table 13 it is auxiliary than particle and dilution fluid viscosity
Increased with extension rate by viscosity known to the above chart and reduced, but is finally 1.45 (to measure water viscosity
1.44), cream is auxiliary also more smaller than smaller particle fluid viscosity under same extension rate.The auxiliary material of viscosity and this particle has close pass
System, the more viscosity of auxiliary material are smaller.
4.3.3pH, data are as shown in table 14:
Each cream of table 14 is auxiliary than particle and dilution pH
The aobvious acidity of purslane-cortex cinnamomi particle as seen from the above table, extension rate increases pH value and also increases, and has with supplementary product consumption
It closes, cream is auxiliary to be increased than small then pH value.Contain oxalic acid, malic acid and amino acid in purslane.Contain cinnamic acid in cortex cinnamomi, is changing
Work, medicine and other aspects have extensive utilization.
The above-mentioned the physical-chemical parameters measurement result of comprehensive analysis and granule evaluation result, cream are auxiliary than particle obtained by 1:3
Parameters are good compared with other proportion performances.
5. the research of anti-ulcerative colitis
5.1 introductions: ulcerative colitis (ulcerative colitis, UC) is a kind of chronic nonspecific intestinal inflammatory
Property disease.It is refractory to be cured since this disease cause of disease and pathogenesis are still not clear, it is easy to recur, the modern times are classified as by the World Health Organization
Difficult treatment.Ulcerative colitis clinical manifestation is abdominal pain, diarrhea, mucus pus and blood stool and tenesmus is characterized.
5.2. laboratory apparatus: the 101 type electric drying oven with forced convection Beijing Yongxing Ke Wei Instrument Ltd., microplate reader,
Multiskan GO)-1510(Thermo Fisher)
5.3. experimental material: (DSS, MP Biomedicals are biomedical public by dextran sulfate sodium MW:36000-50000
Department, lot number: 160110)；Sulfasalazine (the good magnificent medicine company in Shanghai Xinyi, lot number: 09170611).
5.4. experimental animal: male mice 70,20 ± 2g of weight, kind SPF grades of Kunming is purchased from Chengdu up to large experimental animal
Co., Ltd, Quality of Experimental Animals quality certification credit number: SCXK (river) 2015-030.
5.5. experimental group and medication: male Kunming strain mice 70, random point 7 groups, (every group of mouse weight carries out
Variance analysis, P > 0.05) adaptive feeding one week.It is divided into blank group, UC model group, salicylazosulfapyridine group, purslane-meat
Osmanthus particle 1:2,1:3,1:4,1:5 group, naive mice drinking pure water, other mouse drink 7 days induction UC moulds of 5%DSS certainly
Type records the weight of mouse, shit quality, occult blood or hematochezia feature from modeling first day, carry out disease activity according to table 15 and refer to
Number (disease activity index, DAI) scoring.
5% physiological saline of blank group and model group stomach-filling, 1mL/ days；The willow nitrogen of salicylazosulfapyridine group stomach-filling 125mg/kg
Sulfapryidine suspension, 1ml/ days；Each cream of the other stomach-filling 0.12g/ml of grain fraction is auxiliary than particle solution, and equal 1ml/ days.Finally
Stomach-filling is complete, and fasting 1 day.Mouse Colon section about 1cm is taken, 4% paraformaldehyde saves, and uses to H&E pathological section.
DAI=(weight loss scoring+stool scoring+stool blood situation scoring)/3
15 DAI grade form of table
Note: normal stool: forming stool；Loose stool: anus mouth, paste or half form stool are not attached to；Loose stools: can
Attach to dilute watery stool of anus.
5.6. experimental result
5.6.1 DAI appraisal result is shown in Fig. 1: referring to Fig. 1, mouse DAI scoring can be obtained, during preceding modeling in 7 days, mouse
DAI scoring is in rising trend, shows mouse modeling success, the treatment of stomach-filling in the 8th day, and administration group mouse DAI score curve is integrally in
Downward trend, and with the increase of the administration auxiliary ratio of particle cream, mouse DAI scoring decline is smaller, shows purslane-cortex cinnamomi particle pair
UC is effective in cure, and auxiliary than related with the cream for preparing particle.Cream is auxiliary, and than 1:2 and 1:3 group, curative effect is almost the same after treatment 14 days, cream
It is auxiliary more almost the same than 1:4 and 1:5 group curative effect.It can thus be seen that cream it is auxiliary than 1:3 be curative effect ratio node, be higher than 1:3 after curative effect
It significantly reduces, this is auxiliary more closely related than dose reduction after increasing with cream.
5.6.2 H&E coloration result is illustrated in fig. 2 shown below.
Mucous layer, submucosa, lamina propria and placenta percreta constitute enteron aisle.In Fig. 2, A is that complete Colonic Struture Changes are (empty
White group), including mucous layer, submucosa, muscle layer, it is seen that apparent crypts and goblet cell, the intrusion of no inflammation cell.B is 5%
UC model group, segmental colonic mucous layer imperfect fall off crypts and goblet cell reduction, connective tissue loose.C is positive control
Salicylazosulfapyridine group, colon structure is complete, has a large amount of crypts and goblet cell, but mucous layer is said there are also a small amount of inflammatory cell
Bright salicylazosulfapyridine can treat the colitis of DSS induction.D, E, F, G are respectively auxiliary than being 1:2,1:3,1:4,1:5 of cream
Grain group, colonic tissue is complete, has a large amount of crypts and goblet cell, but there are also a small amount of inflammatory cell infiltrations, illustrate each ratio particle
Colonic tissue can be repaired, but the damage for the factor pair colonic tissue that can not all diminish inflammation.Compare that each cream is auxiliary to compare pathological section
Effect can be seen that cream it is auxiliary than 1:3 slice in inflammatory cell number, crypts state and goblet cell form due to other ratios.
Inflammatory cell significantly reduces, and clear in structure complete, crypts is regular.To sum up, the auxiliary granule therapy effect than 1:3 of cream is best.
Summarize: optimization purslane-cortex cinnamomi is with when extraction process by water first, and has prepared purslane cortex cinnamomi granule, and
The physical-chemical parameters of granule are had rated by modernism, while having carried out pharmacology activity research.Show purslane-cortex cinnamomi
Match 12:5, cream it is auxiliary than 1:3 in the case of, obtained granules active is best.
6. the influence that purslane cortex cinnamomi granule adjusts mouse intestinal flora
The variation of 6.1 intestinal flora overall profiles: after DSS modeling and administration, intestinal flora overall profile is had occurred obviously
Variation.Research is using flora OTU quantity and its species cross situation between Wei Entu displaying each group；Using principal component analysis
(PCA) flora abundance entirety difference between displaying sample.
Referring to Fig. 3, normal control, model, positive drug and purslane-cortex cinnamomi each group detect 855 OTU altogether, wherein
Shared OTU only 381, account for 44.6%；Flora uneven class size is larger between illustrating each group, i.e. modeling or administration can significantly change intestines
Road flora OTU composition.From each group-specific OTU quantity, model group is far more than control group and two administration groups, equally
Intestinal inflammatory state after illustrating modeling drives intestinal flora by survival pressure, makes population structure that significant change have occurred
Change；And these specific floras greatly reduce after being administered, and imply the recession of intestinal inflammatory state.
PCA based on sample OTU abundance analysis shows, sample standard deviation can be distinguished obviously between each group, illustrate modeling and administration
Processing generates certain influence to mouse intestinal flora structure, and especially modeling factor influences maximum.Administration group and model group phase
Certain offset, and the trend that oriented Normal group returns has occurred than, intestinal microflora, illustrates that drug treatment can one
Determine the recovery for promoting intestinal microflora in degree.
6.2. the variation of intestinal flora bio-diversity: referring to fig. 4, each group sample interior Analysis of Microbial Diversity table
Bright, modeling processing makes mouse intestinal flora that the increased trend of microbial diversity be presented on the whole, wherein chao1 and PD_
Microbial population abundance is intended to equalize after whole_tree index shows modeling, and rare species abundance can increase；And
Observed_species index equally illustrates that total Number of Species increases.The stabilization of intestinal microecology depends on the ecology of dominant microflora
Occupy-place, to promote symbiosis flora to breed and inhibit the increase of opportunistic pathogenic bacteria or harmful bacteria.In conjunction with the above result shows that, it is small
Mouse intestinal inflammatory state promotes rare flora breeding to accelerate, and the then opposite reduction of dominant microflora abundance, function reduction, the micro- life of enteron aisle
The stability of state is on the hazard.Other than shannon index, on other 3 indexes, positive drug or purslane cortex cinnamomi are given
Medicine group can adjust back diversity indices to a certain extent, show that drug may be by promoting intestinal microflora to restore to mitigate
Intestinal inflammatory.
6.3 intestinal flora species annotation and variance analysis
Each OTU representative series input SILVA database is identified into its biological classification.The result shows that mouse intestinal flora
19 bacterium doors and 168 Pseudomonas can be divided into.In all doors, Bacteroidetes (Bacteroidetes), Firmicutes
(Firmicutes), deferrization bacillus door (Deferribacteres), Proteobacteria (Proeobacteria),
Epsilonbacteraeota and actinomyces door (Actinobacteria) abundance are successively successively decreased, and total amount occupies all bacterium
The 99.89% of group's quantity.In all Pseudomonas, Prevotellaceae_UCG-001 category (Prevost Cordycepps),
Alloprevotella intend prevotella, Bacteroides Bacteroides, Allobaculum other style stick Pseudomonas,
Lachnospiraceae_NK4A136_group belong to (Lachnospira section), Phascolarctobacterium koala Bacillus,
The another branch Pseudomonas of Lactobacillus Lactobacillus, Alistipes, Rikenellaceae_RC9_gut_group category (grind by reason
Cordycepps), Dubosiella category be followed successively by the highest top ten Pseudomonas of abundance identified, abundance is differed from 18.5% to 1.7%.
By Welch's t-test examine, discovery model group actinomyces door abundance compared with control group significantly reduce (by
0.37% is down to 0.21%, P < 0.05), and the bacterium is adjusted back in positive drug combination purslane-cortex cinnamomi group, abundance is respectively
0.31% and 0.43%, and without significant difference between control group.
It is further analyzed using LefSe, compares modeling and purslane-caused intestinal flora abundance of cortex cinnamomi processing exists
Difference in each species taxonomy level.The results show that compared with the control group, modeling processing can cause bifidobacterium family
(Bifidobacteriaceae) 7 points of the up-regulation of 17 categorization levels and red stinkbug Zoopagales (coriobacteriales) etc. such as
The downward of class level.At the same time, compared with model group, purslane-cortex cinnamomi processing group causes Bacteroides
(Bacteroidaceae) 13 points of the up-regulation of 6 categorization levels and bifidobacterium family (Bifidobacteriaceae) etc. such as
The downward of class level.These results indicate that modeling processing causes more bacteria abundance to raise, and purslane-cortex cinnamomi processing is then
Opposite trend is shown, a large amount of floras is caused to be lowered, this adjusting trend may be the original that purslane-cortex cinnamomi plays response to treatment
One of because.In addition, this partial results matches with the variation of aforementioned bacterial diversity.
In the flora of significant difference, compare control group, model group, positive drug group and purslane-cortex cinnamomi group bacterium
Group's abundance finds Bifidobacteriaceae bifidobacterium family, Moryella category, Lachnospiraceae_ referring to Fig. 5
ND_3007_group belong to (Lachnospira section), Bacteroidales_BS11_gut_group section (bacteroid mesh),
Coriobacteriales red stinkbug Zoopagales, Negativibacillus belong to (Firmicutes), Jeotgalicoccus belongs to (grape ball
Cordycepps), Rikenellaceae_RC9_gut_group belong to (reason grinds Cordycepps) etc. and significantly increase or significantly reduce in model group,
And positive drug or purslane-cortex cinnamomi processing can significantly adjust back these floras, as shown in Figure 5, wherein A is
Bifidobacteriaceae (bifidobacterium family) bacterium relative abundance figure in different experiments group；B is that Moryella belongs to bacterium
The relative abundance figure in different experiments group；C is that Lachnospiraceae_ND_3007_group belongs to (Lachnospira section) bacterium not
With relative abundance figure in experimental group；D is Bacteroidales_BS11_gut_group section (bacteroid mesh) bacterium in different realities
Test relative abundance figure in group；E is Coriobacteriales red stinkbug Zoopagales bacterium relative abundance figure in different experiments group；F:
Negativibacillus belongs to (Firmicutes) bacterium relative abundance figure in different experiments group；G is that Jeotgalicoccus belongs to
(staphylococcaceae) bacterium relative abundance figure in different experiments group；H is that Rikenellaceae_RC9_gut_group belongs to (reason
Grind Cordycepps) bacterium relative abundance figure in different experiments group.The present invention provides a kind of systems of anti-ulcerative colitis granule
Preparation Method and its effect.UC mouse is induced with 5%DSS, is scored from DAI, colonic tissue morphosis shows purslane-cortex cinnamomi
Grain has therapeutic effect to UC mouse.Variation from intestinal flora profile and flora bio-diversity and flora species annotation and poor
Different analysis result illustrates that drug treatment can promote the recovery of intestinal microflora to a certain extent, and drug may be by promoting intestines
Road Bacterial community restores to mitigating intestinal inflammatory, from LefSe analysis as the result is shown some floras of bifidobacterium family to UC mouse
Intestinal flora has adjustment effect.
Claims (10)
1. a kind of anti-ulcerative colitis granule, it is characterised in that: the anti-ulcerative colitis granule is by purslane
And cortex cinnamomi is prepared；The starting weight of the purslane and cortex cinnamomi ratio is 10-14:3-7.
2. anti-ulcerative colitis granule according to claim 1, it is characterised in that: the weight of the purslane and cortex cinnamomi
Measuring ratio is 11-12:4-5.
3. a kind of preparation method of anti-ulcerative colitis granule, it is characterised in that: the anti-ulcerative colitis granule
Preparation method the following steps are included:
1) it is that the ratio of 10-14:3-7 weighs purslane and cortex cinnamomi and mixes purslane with cortex cinnamomi by weight, is mixed
Close object；
2) water is added in the obtained mixture of step 1), 60 DEG C -100 DEG C at a temperature of extract 1-6h, filter the dregs of a decoction, it is dense
It is reduced to thick medicinal extract；
3) thick medicinal extract that step 2) is prepared was placed in -80 DEG C of refrigerator freezings not less than 24 hours, and it is dry that taking-up is placed on freezing
It is dry in dry device, it is ground into fine powder after the completion；
4) fine powder for taking step 3) to be prepared is mixed with sucrose；The weight ratio of the fine powder and sucrose is 1:2-5；
5) 80% ethanol solution is added into the mixture that step 4) is prepared, hand is made and pinches the i.e. scattered softwood of agglomerating, flicking；
6) particle is made by No. 1 medicine sieve in a manner of squeezing granulation in the softwood that step 5) is prepared.
4. the preparation method of anti-ulcerative colitis granule according to claim 3, it is characterised in that: the step 1)
In, the weight ratio of purslane and cortex cinnamomi is 11-12:4-5.
5. the preparation method of anti-ulcerative colitis granule according to claim 4, it is characterised in that: the step 2)
The solid-liquid ratio of middle mixture and water is 1:10-50.
6. the preparation method of anti-ulcerative colitis granule according to claim 5, it is characterised in that: the step 2)
The solid-liquid ratio of middle mixture and water is 1:40.
7. the preparation method of anti-ulcerative colitis granule according to claim 6, it is characterised in that: the step 2)
Extracting temperature be 90 DEG C, extraction time is 5h.
8. the preparation method of anti-ulcerative colitis granule according to claim 6, it is characterised in that: the step 3)
The time of middle drying is for 24 hours.
9. according to the preparation method of anti-ulcerative colitis granule described in claim 3-8 any claim, feature
Be: in the step 4), the weight ratio of fine powder and sucrose is not less than 1:3.
10. the preparation method of anti-ulcerative colitis granule according to claim 9, it is characterised in that: described anti-routed
The preparation method of ulcer colitis granule is after step 6) further include:
7) particle that step 6) is prepared is put into baking oven dry, screens out big particle with No. 1 after drying, No. 4 screen out
Fine powder is removed, finished product is obtained.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1785389A (en) * | 2005-11-18 | 2006-06-14 | 李胜利 | Preparation method of Chinese medicine for treating ulcero proctitis |
| CN101837096A (en) * | 2010-05-31 | 2010-09-22 | 牛菊萍 | Enema medicament for curing ulcerative colitis |
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2018
- 2018-09-11 CN CN201811057108.9A patent/CN109157562A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1785389A (en) * | 2005-11-18 | 2006-06-14 | 李胜利 | Preparation method of Chinese medicine for treating ulcero proctitis |
| CN101837096A (en) * | 2010-05-31 | 2010-09-22 | 牛菊萍 | Enema medicament for curing ulcerative colitis |
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| Title |
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| 刘力 等: "马齿苋对溃疡性结肠炎小鼠结肠黏膜病理形态的影响", 《山西中医》 * |
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