CN106668394A - Pharmaceutical composition for preventing and treating rheumatoid arthritis and preparation method thereof - Google Patents
Pharmaceutical composition for preventing and treating rheumatoid arthritis and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a pharmaceutical composition for preventing and treating rheumatoid arthritis and a preparation method of the pharmaceutical composition. The pharmaceutical composition is prepared by matching the crude drugs of anemone rivularis, rose apple, iris speculatrix root, himalayan maple and fimbristylis dichotomacan, can be prepared into various preparations through the conventional preparation technology, and has remarkable effects of preventing and treating osteoproliferation.
Description
Technical field
The invention belongs to technical field of traditional Chinese medicines, more particularly to a kind of pharmaceutical composition for preventing and treating rheumatoid arthritis and its
Preparation method.
Background technology
Rheumatoid arthritis（RA）It is a kind of autoimmune disease being characterized with chronic symmetry arthritis, its base
This lesion is synovial membrane inflammation, can cause cartilage destruction and bone erosion, causes joint deformity and dysfunction.Rheumatism association of the U.S.
RA treatment updated Guidelines can be issued in 2002, has proposed that RA therapeutic strategies are to prevent disease development and reach alleviation,
Suppress synovial membrane inflammation, prevent and control destruction of joint, prevent function from losing and mitigate pain.CIA models are international at present used
RA animal models, with the clinic more similar to mankind RA, histopathology and immunological changes feature.It is known
The joint injury and osteoclasia of RA are mediated by chronic synovitis disease.The external irritant such as infection or wound, starts itself and exempts from
Epidemic disease is reacted, and synovial cell is bred rapidly, and various inflammatory cells are strengthened the infiltration of synovial membrane, at the same secrete produce a large amount of cells because
Son, inflammatory mediator and various antibody etc., combine and have mediated RA inflamed joints to damage and osteoclasia.
The traditional Chinese medical science understanding with treat this disease on have original advantage.Rheumatic arthritis belongs to Chinese medical ' arthralgia syndrome ' category, goes through
" severe and migratory arthralgia ", " arthroncus of knee ", " arthritis ", " heumatism ", " rising sun numbness " etc. are also referred to as according to its feature for doctor.The traditional Chinese medical science thinks this disease
Weak by ferritic, positive unsaturated vapor, knee manages not close, weakened defensive QI, in addition diseases caused by external factors wind, cold, wet, heat evil and formed.I.e.《Element
Ask numbness opinions》Say that " wet three gas of chill is miscellaneous extremely, combined into numbness." QI-blood circulation is smooth, addiction blood resistance network is the center ring of bi Zheng
Section.Sick to be taken good care of oneself accidentally more because natural endowment element is empty, disease and evil is felt again, the heresy of direct feeling rheumatic fever, or chill wet three is evil, also can be by dirty
Caused by internal organs functional disturbance, such as positive hot constitution, or cloudy anemia consumption.What immortality etc. thinks that the interpretation of the cause, onset and process of an illness key of RA morbidities is that heresy is orthogonal and strives
In positive wound invaded with heresy.Positive wound refers to that the healthy tendency virtual loss and blood stasis, the stagnation of the circulation of vital energy, phlegm of human body are coagulated, and is the internal factor of its morbidity；Heresy is invaded
Duties is damp and hot in chill more.
Portugal：This product is the dry fruit of myrtle Hainan Pu peach Syzygium cumini (L.) Skeels.Autumn adopts
Collection, removes branches and leaves, dries.【Nature and flavor】Puckery, bitter, temperature is pungent.【Function with cure mainly】Warm kidney dispelling cold.For " three is evil " disease, kidney is cold sick,
Stranguria with turbid discharge.【Proterties】This product is rounded, oblique square is circular, 1~2cm, 0.5~1cm wide long.Surface sepia to black, exocarp wrinkles
Shorten netted line into, exocarp comes off sometimes, and surface is more smooth, and endocarp is fallow, top carries place calyx, like bottleneck shape, in
The withered style of the heart visible, 1 piece of seed, yellowish-brown is hard.Gas delicate fragrance, it is lightly seasoned.Record and issue Tibetan medicine standard in the Ministry of Public Health（The
One）, standard number：WS3-BC-0112-95.
Anemone rivularis：This product is cohosh anemone rivularis Anemone rivularis Buch.-Ham.ex DC. and belongs to together
The dry fruit of various plants.Ripening fruits is gathered after autumn, net impurity is picked, and is dried.【Nature and flavor】Pungent, hardship, temperature.【Function with cure mainly】
Corruption is gone, stomach temperature, drainage yellow water is lifted.For mawworm, shouting pain, the diseases such as yellow water are accumulated in snake bite, cold tumour, gonorrhoea, joint.
【Proterties】This product is in elongated oval, 5~8mm, about 2mm wide long.The aobvious light green color of surface chartreuse or one end, smooth, base portion
Tool short handle, tip harbors elongated style, in hook-shaped bending.Matter is tough, pericarp with plant skin separate, section milky, or in shallow brown
Color, shows slightly oiliness.Gas is micro-, and taste is micro- peppery.Record and issue Tibetan medicine standard in the Ministry of Public Health（First）, standard number：WS3-BC-0066-
95。
Dichotomous fimbristylis herb：It is Cyperaceae genus fimbristylis plant two qi dichotomous fimbristylis herb Fimbristylis dichotoma (L) Vahl
[Scirpus dichotomus L.;S.annuus All.;Fimbristylis annua(All.)Roem.et Schult]
Herb.Summer, autumn harvesting, clean, and dry.【Nature and flavor】It is lightly seasoned；It is cold in nature.【Return through】Kidney；Bladder warp.【Indication】Heat-clearing profit
Urine；Removing toxic substances.Main difficult urination；Damp and hot edema；Gonorrhoea；Infant fetal toxin.【Chemical analysis】Containing dihydrocyperaquinone
(dihydrooyperaqnone), tetrahydrocyperaquinone.(tetrahydrocyperaquinone), cyperaquinone
(cyperapuinone), hydroxyl cyperaquinone (hydroxycyperapuinone), removes first cyperaquinone
(demethylcyperapuinone).【Former phytomorph】Not tally dichotomous fimbristylis herb, draft.Stalk is grown thickly, 20-50cm high.Complete stool without
Hair has thin pubescence.Leaf is linear, is shorter than stalk, and 1-2.5mm wide, tip is suddenly sharp or blunt；Sheath base portion keratin.Lobate bract under inflorescence
3-4, normal 1-2 pieces, is longer than inflorescence.Cyme is appeared again or simply；Small ear is avette or round shape long is avette, 5-12mm long, 2- wide
3mm, there is most flowers；Scale is avette or Long Circle, 2-2.5mm long, and sepia is glossy, there is 3-5 arteries and veins, and tip has short point；Stamen
2-3；Style is flat, and echinid is arranged at top；Column cap 2.Pyrene obovate wide, biconvex shape, 1-1.2mm long, white to filbert,
Surface is grown crosswise circular reticulate pattern, and longitudinal rib 7-9 bars, significant protuberance has brown short handle.Flower, the fruiting period 7-10 months.Record in the big diction of Chinese medicine
Allusion quotation.
Little Hua orrisroots：For Iridaceae iris little Hua iris Iris speculatrix Hance rhizome and
Root.The 9-11 months harvest, and clean segment, dry.【Nature and flavor】It is pungent；Temperature；It is slightly poisonous.【Indication】Invigorating blood circulation analgesia；Dispelling wind and eliminating dampness.It is main
Traumatic injury；Wind-cold-dampness arthralgia；Mad dog is bitten；Snakebite.【Former phytomorph】Little Hua iris perennial herbs, 30-40cm high.Plant
Strain base portion is with the old leaf sheath fiber and the sheath shape leaf by aciculiform of sepia.Rhizome grows wild, more sturdy, and link is significant.Basal leaf
Sword-shaped or bar shaped, 10-40cm long, 3-8mm wide, tip are tapering, base portion sheath shape, full edge；Stem leaf 3-4 pieces, it is short compared with basal leaf.Flower
Stem is flat, there is bract 2, lanceolar；Include l-2 flowers；Bennet 3-5.5cm long, bends after blossom fall；Flower bluish violet or light blue, directly
Footpath 5-6cm, foreign steamer sliver cochlear, the annular patch for having darkviolet, the orange-yellow palpus shape adjunct of the close tool of middle arteries, lubrication groove sliver drapes over one's shoulders pin
Shape, inclines；Stamen 3, flower pesticide white；Ovary is the next, and Room 3, style branch is flat, and 3 branches, petal-shaped, tip 2 splits, and edge has
Tooth.Capsule is oval, 5-5 long, 5cm, diameter l-1.5cm, and tip has thin styloid beak, and lobe is 3 valves when ripe.Seed pears
Shape, sepia.The month at florescence 4-5, the fruiting period 7-8 months.Record in dictionary of medicinal plant.
Moth tree：Pueraria lobota trailing plants maple Acer grosseri Pax var.hersii are split for Aceraceae Acer plants are long（Rehd.）
Rehd. the spray and fruit of ［ A.hersii Rehd. ］.9-10 month picking fruits, dry；The 6-7 months harvest spray, dry.【Property
Taste】It is bitter；It is salty；It is flat.【Indication】Cough-relieving；Sore.The new chronic cough of master is coughed；Thrush.【Original shape state】It is tall that length splits Pueraria lobota trailing plants maple fallen leaves
Wood, up to 8m.Bark is filbert, smooth；Annual shoot green or purple green, perennial branch lark or taupe, leaf opposite；
Petiole 2-3cm long, it is thin thin, without hair；Blade papery, avette, 7-9cm long, 5-6cm wide, edge has close and sharp heavy sawtooth, first
Hold sharp point, base portion to be bordering on heart-shaped, often deeper 3 and split, central sliver is larger, and triangular shape is avette, above bottle green, without hair；Under
Face light green, is then come off always in vein base portion when tender by faint yellow feathering；Base goes out 3, arteries and veins, lateral vein pinniform.Flower unisexuality, male and female are different
Strain, often into thin thin sagging raceme；Sepal 5；Petal 5；Stamen 8, without hair, the agensis in female flower；Floral disc is located at stamen
Inner side；Ovary purple, the agensis in male flower.Bennet 3-4mm long.Lavender, ripe rear yellowish-brown during samara children；Pyrene is micro- flat
Flat, together with pyrene 2.5-2.9cm long, about 5mm wide or is bordering on level at flare up obtuse angle to wing.April at florescence, fruiting period September.Record
In dictionary of medicinal plant.
The content of the invention
The purpose of the present invention is to overcome the shortcomings of background technology, there is provided a kind of medicine for effectively preventing and treating rheumatoid arthritis
Composition and preparation method thereof.
The present invention adopts the following technical scheme that realization：
The composition and weight portion for being made the bulk drug of the pharmaceutical composition for preventing and treating rheumatoid arthritis be：
Anemone rivularis 342-348 weight portion Portugal 312-318 weight portion little Hua orrisroot 124-128 weight portion moth trees 112-
116 weight portion dichotomous fimbristylis herb 105-109 weight portions.
The pharmaceutical composition for preventing and treating rheumatoid arthritis is preferably used in, is made up of the bulk drug of following weight portion：
The weight portion dichotomous fimbristylis herb of 345 weight portion Portugal of anemone rivularis, 315 weight portion little Hua orrisroots, 126 weight portion moth tree 114
107 weight portions.
A kind of pharmaceutical composition for preventing and treating rheumatoid arthritis, it is characterised in that pharmaceutical composition can use galenic pharmacy
Conventional method prepare piece agent or capsule or dripping pill.
A kind of pharmaceutical composition for preventing and treating rheumatoid arthritis, it is characterised in that pharmaceutical composition and chemical drugs or Chinese medicine
The preventing and treating medicine for treating rheumatoid arthritis of composition.
A kind of preparation method of the pharmaceutical composition for preventing and treating rheumatoid arthritis, it is characterised in that make as follows
It is standby：
The composition and weight portion of bulk drug be：Anemone rivularis 342-348 weight portion Portugal 312-318 weight portion little Hua orrisroots
124-128 weight portion moth tree 112-116 weight portion dichotomous fimbristylis herb 105-109 weight portions；
Preparation method：
（1）Anemone rivularis, Portugal, little Hua orrisroots, moth tree, dichotomous fimbristylis herb are taken by bulk drug proportioning, is mixed, it is dense with percentage by weight
16% ethanol being spent as solvent, being taken in 37 DEG C of temperature extractions, extraction time is 18 times, and each extraction time is 65 hours, each solvent
Consumption is 44 times of bulk drug gross weight, is filtered, and obtains dregs of a decoction A and extract solution A, and extract solution A reclaims ethanol, is concentrated into relative density
1.08, filtration, liquid is first washed with water by SP70 large pore resin absorption columns, then with the ethanol solution of weight percent concentration 32%
Wash-out SP70 large pore resin absorption columns, collect the ethanol eluate of weight percent concentration 32%, reclaim ethanol, and concentrate drying is obtained final product
Extract A；
（2）Take step（1）Dregs of a decoction A, with the ethanol of weight percent concentration 48% as solvent, heating and refluxing extraction 31 times is carried every time
The time is taken for 0.3 hour, each solvent load is 78 times of dregs of a decoction A weight, and filtration obtains dregs of a decoction B and extract solution B, and extract solution B is returned
Ethanol is received, relative density 1.12 is concentrated into, filtered, liquid is first washed with water, then use by SIP1200 large pore resin absorption columns
The ethanol solution of weight percent concentration 86% elutes SIP1200 large pore resin absorption columns, collects the ethanol of weight percent concentration 86%
Eluent, reclaims ethanol, and concentrate drying obtains final product extract B；
（3）Extract A and extract B are mixed, pharmaceutical composition is obtained final product.
Preferred a kind of preparation method of the pharmaceutical composition for preventing and treating rheumatoid arthritis, it is characterised in that by following step
It is rapid to prepare：
The composition and weight portion of bulk drug be：The weight portion of 345 weight portion Portugal of anemone rivularis, 315 weight portion little Hua orrisroots 126
The weight portion of 114 weight portion dichotomous fimbristylis herb of moth tree 107；
Preparation method：
（1）Anemone rivularis, Portugal, little Hua orrisroots, moth tree, dichotomous fimbristylis herb are taken by bulk drug proportioning, is mixed, it is dense with percentage by weight
16% ethanol being spent as solvent, being taken in 37 DEG C of temperature extractions, extraction time is 18 times, and each extraction time is 65 hours, each solvent
Consumption is 44 times of bulk drug gross weight, is filtered, and obtains dregs of a decoction A and extract solution A, and extract solution A reclaims ethanol, is concentrated into relative density
1.08, filtration, liquid is first washed with water by SP70 large pore resin absorption columns, then with the ethanol solution of weight percent concentration 32%
Wash-out SP70 large pore resin absorption columns, collect the ethanol eluate of weight percent concentration 32%, reclaim ethanol, and concentrate drying is obtained final product
Extract A；
（2）Take step（1）Dregs of a decoction A, with the ethanol of weight percent concentration 48% as solvent, heating and refluxing extraction 31 times is carried every time
The time is taken for 0.3 hour, each solvent load is 78 times of dregs of a decoction A weight, and filtration obtains dregs of a decoction B and extract solution B, and extract solution B is returned
Ethanol is received, relative density 1.12 is concentrated into, filtered, liquid is first washed with water, then use by SIP1200 large pore resin absorption columns
The ethanol solution of weight percent concentration 86% elutes SIP1200 large pore resin absorption columns, collects the ethanol of weight percent concentration 86%
Eluent, reclaims ethanol, and concentrate drying obtains final product extract B；
（3）Extract A and extract B are mixed, pharmaceutical composition is obtained final product.
A kind of preparation method of the pharmaceutical composition for preventing and treating rheumatoid arthritis, it is characterised in that pharmaceutical composition can be with
Piece agent or capsule or dripping pill are prepared using the conventional method of galenic pharmacy.
A kind of preparation method of the pharmaceutical composition for preventing and treating rheumatoid arthritis, it is characterised in that pharmaceutical composition and change
Learn medicine or Chinese medicine composition preventing and treating medicine for treating rheumatoid arthritis.
Pharmaceutical composition preventing and treating Rheumatoid Arthritis are notable.
Specific embodiment
Embodiment 1：Prevent and treat pharmaceutical composition of rheumatoid arthritis and preparation method thereof
The composition and weight portion for preventing and treating the bulk drug of the pharmaceutical composition of rheumatoid arthritis be：Anemone rivularis 345g Portugals
315g little Hua orrisroot 126g moth tree 114g dichotomous fimbristylis herbs 107g；
Preparation method：
（1）Anemone rivularis, Portugal, little Hua orrisroots, moth tree, dichotomous fimbristylis herb are taken by bulk drug proportioning, is mixed, it is dense with percentage by weight
16% ethanol being spent as solvent, being taken in 37 DEG C of temperature extractions, extraction time is 18 times, and each extraction time is 65 hours, each solvent
Consumption is 44 times of bulk drug gross weight, is filtered, and obtains dregs of a decoction A and extract solution A, and extract solution A reclaims ethanol, is concentrated into relative density
1.08, filtration, liquid is first washed with water by SP70 large pore resin absorption columns, then with the ethanol solution of weight percent concentration 32%
Wash-out SP70 large pore resin absorption columns, collect the ethanol eluate of weight percent concentration 32%, reclaim ethanol, and concentrate drying is obtained final product
Extract A；
（2）Take step（1）Dregs of a decoction A, with the ethanol of weight percent concentration 48% as solvent, heating and refluxing extraction 31 times is carried every time
The time is taken for 0.3 hour, each solvent load is 78 times of dregs of a decoction A weight, and filtration obtains dregs of a decoction B and extract solution B, and extract solution B is returned
Ethanol is received, relative density 1.12 is concentrated into, filtered, liquid is first washed with water, then use by SIP1200 large pore resin absorption columns
The ethanol solution of weight percent concentration 86% elutes SIP1200 large pore resin absorption columns, collects the ethanol of weight percent concentration 86%
Eluent, reclaims ethanol, and concentrate drying obtains final product extract B；
（3）Extract A and extract B are mixed, pharmaceutical composition is obtained final product.
Embodiment 2：Prevent and treat pharmaceutical composition of rheumatoid arthritis and preparation method thereof
The composition and weight portion for preventing and treating the bulk drug of the pharmaceutical composition of rheumatoid arthritis be：Anemone rivularis 342g Portugals
318g little Hua orrisroot 124g moth tree 116g dichotomous fimbristylis herbs 105g；
Preparation method：
（1）Anemone rivularis, Portugal, little Hua orrisroots, moth tree, dichotomous fimbristylis herb are taken by bulk drug proportioning, is mixed, it is dense with percentage by weight
16% ethanol being spent as solvent, being taken in 37 DEG C of temperature extractions, extraction time is 18 times, and each extraction time is 65 hours, each solvent
Consumption is 44 times of bulk drug gross weight, is filtered, and obtains dregs of a decoction A and extract solution A, and extract solution A reclaims ethanol, is concentrated into relative density
1.08, filtration, liquid is first washed with water by SP70 large pore resin absorption columns, then with the ethanol solution of weight percent concentration 32%
Wash-out SP70 large pore resin absorption columns, collect the ethanol eluate of weight percent concentration 32%, reclaim ethanol, and concentrate drying is obtained final product
Extract A；
（2）Take step（1）Dregs of a decoction A, with the ethanol of weight percent concentration 48% as solvent, heating and refluxing extraction 31 times is carried every time
The time is taken for 0.3 hour, each solvent load is 78 times of dregs of a decoction A weight, and filtration obtains dregs of a decoction B and extract solution B, and extract solution B is returned
Ethanol is received, relative density 1.12 is concentrated into, filtered, liquid is first washed with water, then use by SIP1200 large pore resin absorption columns
The ethanol solution of weight percent concentration 86% elutes SIP1200 large pore resin absorption columns, collects the ethanol of weight percent concentration 86%
Eluent, reclaims ethanol, and concentrate drying obtains final product extract B；
（3）Extract A and extract B are mixed, pharmaceutical composition is obtained final product.
Embodiment 3：Prevent and treat pharmaceutical composition of rheumatoid arthritis and preparation method thereof
The composition and weight portion for preventing and treating the bulk drug of the pharmaceutical composition of rheumatoid arthritis be：Anemone rivularis 348g Portugals
312g little Hua orrisroot 128g moth tree 112g dichotomous fimbristylis herbs 109g；
Preparation method：
（1）Anemone rivularis, Portugal, little Hua orrisroots, moth tree, dichotomous fimbristylis herb are taken by bulk drug proportioning, is mixed, it is dense with percentage by weight
16% ethanol being spent as solvent, being taken in 37 DEG C of temperature extractions, extraction time is 18 times, and each extraction time is 65 hours, each solvent
Consumption is 44 times of bulk drug gross weight, is filtered, and obtains dregs of a decoction A and extract solution A, and extract solution A reclaims ethanol, is concentrated into relative density
1.08, filtration, liquid is first washed with water by SP70 large pore resin absorption columns, then with the ethanol solution of weight percent concentration 32%
Wash-out SP70 large pore resin absorption columns, collect the ethanol eluate of weight percent concentration 32%, reclaim ethanol, and concentrate drying is obtained final product
Extract A；
（2）Take step（1）Dregs of a decoction A, with the ethanol of weight percent concentration 48% as solvent, heating and refluxing extraction 31 times is carried every time
The time is taken for 0.3 hour, each solvent load is 78 times of dregs of a decoction A weight, and filtration obtains dregs of a decoction B and extract solution B, and extract solution B is returned
Ethanol is received, relative density 1.12 is concentrated into, filtered, liquid is first washed with water, then use by SIP1200 large pore resin absorption columns
The ethanol solution of weight percent concentration 86% elutes SIP1200 large pore resin absorption columns, collects the ethanol of weight percent concentration 86%
Eluent, reclaims ethanol, and concentrate drying obtains final product extract B；
（3）Extract A and extract B are mixed, pharmaceutical composition is obtained final product.
Embodiment 4：The preparation of tablet
The pharmaceutical composition 677g of Example 1, adds starch 179g, mixes, and granulation is dried, plus microcrystalline cellulose 30g, stearic
Sour magnesium 6g, mixes, and is pressed into 3500, obtains final product medicinal composition tablets.
Embodiment 5：The preparation of capsule
The pharmaceutical composition 636g of Example 2, adds starch 257g, mixes, and granulation is dried, whole grain, adds appropriate stearic acid
Magnesium, mixes, and encapsulated 2800, obtains final product medicament composition capsule.
Embodiment 6：The preparation of dripping pill
Weigh (80 DEG C) heating of Macrogol 6000 167g water-baths and boil molten, add the pharmaceutical composition 11g of embodiment 3, fully stir
Mix uniform, with atoleine as cooling agent, put glass tube（4*80cm）In, chilling temperature is 5 DEG C, and drip internal-and external diameter is 7.0/
2.0 (mm/mm), drip is 2.4cm away from liquid level, and drop speed is optimum condition with every point 57 drop, and the cold of dripping pill surface is blotted with cotton
Solidifying agent, obtains final product medicament composition dropping pills.
Embodiment 7：Prevent and treat the pharmaceutical composition of rheumatoid arthritis
The composition and weight portion for preventing and treating the bulk drug of the pharmaceutical composition of rheumatoid arthritis be：
The weight portion of 110 180 weight portion Portugal of weight portion moth tree of little Hua orrisroots 320.
Embodiment 8：Prevent and treat the pharmaceutical composition of rheumatoid arthritis
The composition and weight portion for preventing and treating the bulk drug of the pharmaceutical composition of rheumatoid arthritis be：
The weight portion of 100 150 weight portion dichotomous fimbristylis herb of weight portion moth tree of little Hua orrisroots 110.
Embodiment 9：Prevent and treat the pharmaceutical composition of rheumatoid arthritis
The composition and weight portion for preventing and treating the bulk drug of the pharmaceutical composition of rheumatoid arthritis be：
The weight portion of 140 100 weight portion dichotomous fimbristylis herb of weight portion moth tree of little Hua orrisroots 100.
Experimental example 1：Prevent and treat the experimental study of rheumatoid arthritis
1 experiment material
1.1 animal
Healthy male Wistar rat, cleaning grade, weight (162 ± 10) g, Academy of Military Medicine, PLA's reality
Test animal center and (credit number is provided：SCXK- (army) 2002-001).
1.2 medicines and reagent
Pharmaceutical composition（The pharmaceutical composition of embodiment 1）With the decocting liquid for being preceding made medicinal material containing 0.1g/mL.Collagen Type VI (the C of ox II
II), chondrex Products；Complete Freund's adjuvant (CFA), Difco Products；The β of interleukins (IL) -1, tumour are bad
Necrosis factor (TNF)-α and IL-6 radio immunoassays (RIA) determine kit, Chinese People's Liberation Army General Hospital's scientific and technological development
Put and exempt from institute's product in center；Prostaglandin (PG) E2Enzyme linked immunosorbent assay detection kit, U.S. R＆D imports packing product；
Rabbit Anti-Rat IgM (bsb-0346R)、Rabbit Anti-Rat IgG (bsb-0293R)、streptavidin/
HRP (bsb-0437R), Beijing Bo Aosen Bioisystech Co., Ltd product.
1.3 instrument
BMJ-1 biological tissue embeddings machine, QPJ-1C paraffin slicing machines (Tianjin Aviation Mechano-Electrical Co.)；111N-101-20g digital displays are swum
Mark slide calliper rule (wide land numeral observing and controlling limited company)；(Planmed Oy are public for Finland for LANMED SOPHIE breast molybdenum targets machine
Department)；GC911 radio-immunities gamma counter (science and technology industry parent company of Chinese University of Science and Technology)；Multiskan Ascent ELIASAs
(Labsystems companies of Finland).
2 experimental techniques
2.1 modelings and administration
Rat is randomly divided into normal group, model group and pharmaceutical composition group, every group 10.In addition to normal group, remaining 2 groups of preparation
CIA models：The appropriate emulsifying agents of ox C II are taken, 0.1mol/L acetic acid (4 DEG C overnight) is dissolved in, CFA is added, suspension emulsion is fully newborn
Change, be degree to instill in water in pearl, take the appropriate emulsion for preparing, every rat is with 0.1mL (containing 100 gC II) in root of the tail portion
Subcutaneous scatterplot injection.Equivalent emulsifying agent on the 7th excites immune.The 7th day after modeling starts gastric infusion, and pharmaceutical composition is to medicament
Amount is respectively 0.2g medicinal materials/kg, is administered by 1mL/100g weights, and one time a day, continuous 4 weeks.Meanwhile, normal group and model group
Gavage isometric(al) distilled water.
2.2 materials and tissue treatment
Rat eye rear vein beard on the 35th takes blood after immune, and 4 DEG C of centrifugations take -20 DEG C of preservations of serum stand-by；Double hind leg knees are taken to close
Ankle-joint 4% paraformaldehyde solution, 4 DEG C of fixations after section and the right side, after x-ray film making (side position), 10% disodium EDTA solution
Decalcification, FFPE, the m of conventional section 5, Hematoxylin-eosin (HE) dyeing；Carried out after taking left back ankle-joint peeling, liquid nitrogen frozen
Tissue homogenate, takes -20 DEG C of preservations of supernatant stand-by.
2.3 Testing index
2.3.1 the incidence of disease and arthritis index score
Since immune 8d, a Clinical points are carried out per 3d, per 4 points of systems of foot, totally 16 points.Using 0~4 grade of The joint evaluation method.0
Point, without arthritis：1 point, small toe joint mild swelling；2 points, small toe joint and pedal swelling；3 points, the sufficient pawl below ankle-joint
Swelling；4 points, including whole arthroncus including ankle-joint.Arthritis Incidence：Sentenced with least 1 pawl Clinical points >=2 point
Break as arthritis occurs.
2.3.2 joint tissue PATHOMORPHOLOGICAL OBSERVATION OF PULLORUM and scoring
Light Microscopic observation ankle-joint synovial membrane, cartilage, the pathological change of bone, and broken from inflammatory cell infiltration, pannus formation, cartilage
The aspects such as bad and bone erosion carry out semi-quantitative analysis.Standards of grading are with reference to pertinent literature.Inflammatory cell infiltration：It is 0 point, thin without inflammatory
Born of the same parents；It is 1 point, sparse to be dispersed in；2 points, comparatively dense；3 points, largely.Pannus is formed：0, without formation；1 point, there is ＜ 50% on a small quantity；2
Point, pannus area ＞ 50%；3 points, largely exist.Articular cartilage damage：0 point, without destruction；1 point, special mess shape destruction ＜ 1/3；2
Point, 1/2 destruction；3 points, 2/3 destruction；4 points, all destruction.Bone erosion degree：It is 0 point, non-corrosive；It is 1 point, a small amount of to corrode ＜ 50%；
2 points, erosion area ＞ 50%；It is 3 points, a large amount of to corrode.
2.3.3 joint X-ray is analyzed
Score standard.Bone erosion：0 point, normally；It is 1 point, suspicious or have 1 small local lesion；2 points, 2 local lesions or disease
Stove total surface area≤50%；3 points, the focus or focus total surface area ＞ 50% of more than 3；4 points, articular surface is destroyed completely, or is gone out
Existing bone reduces.Joint space：0 point, normally；It is 1 point, suspicious or have 1 small local lesion；2 points, joint occurs narrow；3 points,
Arthrocleisis or joint space are broadening.Osteoclasia：0 point, normally；1 point, osteoporosis phase；2 points, destruction of bone phase；3 points, sclerotin
The heavy damage phase；It is 4 points, tetanic.
2.3.4 proinflammatory cytokine and inflammatory mediator content
RIA detects the content of proinflammatory cytokine IL-1 β, IL-6 and TNF-α in serum and joint homogenates supernatant, enzyme-linked
The content of immunoadsorbent technics detection serum PG E2, concrete operations are carried out by kit specification.
3 statistical methods
Using SPSS15.0 softwares, the evaluation statistical method of histopathology and iconography score uses non-parametric test
(Kruskal-Wallis test), morbidity statistics uses Chi-square Test, and remaining uses the multifactor analysis of variance.Experimental data is with x
± s is represented.P ＜ 0.05 are that difference is statistically significant.
4 results
Influence of 4.1 pharmaceutical compositions to collagen induced-arthritis rat arthritis Clinical points and the incidence of disease
Normal rats joint deforms without redness；Model group rats osteoarthritis clinical symptom starts on the 10th after immune
Existing, 17 d are red and swollen substantially to be aggravated, 8.2 points of highest Clinical points, the partially visible dysarthrasises of 28 d.The incidence of disease is from after being immunized
23 d play the high level for maintaining 90% always, and 32 d reach 100%.Pharmaceutical composition group can substantially reduce Clinical points
（3.0 point）And the incidence of disease（35%）.
Influence of 4.2 pharmaceutical compositions to collagen induced-arthritis rat pathological change
Normal group articulation structure is normal, and synovial tissue is formed without hyperplasia without congestion and edema, synovial cell without pannus, and joint is soft
Bone surface is smooth, nothing is stripped off, and bone trabecula size, arrangement are normal under articular cartilage；The visible obvious synovial cell proliferation of model group,
Arrangement disorder, synovial tissue's congestion and edema, and visible inflammatory cell infiltration, pannus are formed, cartilage cells of superficial layer denaturation, it is bad
Extremely, articular cartilage face is stripped off, and is had in articular cavity and substantial amounts of is stripped off slough, subchondral bone girder arrangement disorder；Drug regimen
Wu Zu synovial tissues congestion and edema substantially mitigates, and synovial cell proliferation substantially mitigates, and inflammatory cell infiltration is substantially reduced, and blood vessel increases
Raw and pannus is formed and is suppressed.
Influence of 4.3 pharmaceutical compositions to collagen induced-arthritis rat Imageology
Normal group articular surface of ankle is smooth, and joint space is clear；Model group articular surface is destroyed in worm-eaten sample, marginality bone erosion,
With osteoproliferation reaction, epiphysis hyperplasia, thick, joint space is narrow or broadening, and even bony ankylosis is mutually merged in joint；With mould
Type group is compared, and pharmaceutical composition group articular surface is more smooth, and joint space is more visible.
4.4 pharmaceutical compositions are situated between to collagen Ⅱ induced rat blood serum and joint tissue proinflammatory cytokine and inflammation
The influence of matter
In model group serum and inflamed joint tissue in proinflammatory cytokine IL-1 β, TNF-α and IL-6 contents and serum
The content of inflammatory mediator PGE2 is significantly raised compared with normal group, and difference is statistically significant (P ＜ 0.05, P ＜ 0.01).Except pass
In section tissue outside TNF-α, pharmaceutical composition group can significantly reduce increase in CIA rat blood serums and inflamed joint proinflammatory
The content (P ＜ 0.05, P ＜ 0.01, P ＜ 0.001) of cell factor IL-1 β, TNF-α and IL-6 and inflammatory mediator PGE2.
Claims (8)
1. a kind of pharmaceutical composition for preventing and treating rheumatoid arthritis, it is characterised in that be made the bulk drug of the pharmaceutical composition
Composition and weight portion are：
Anemone rivularis 342-348 weight portion Portugal 312-318 weight portion little Hua orrisroot 124-128 weight portion moth trees 112-
116 weight portion dichotomous fimbristylis herb 105-109 weight portions.
2. a kind of pharmaceutical composition for preventing and treating rheumatoid arthritis according to claim 1, it is characterised in that be made the medicine
The composition and weight portion of the bulk drug of compositions be：
The weight portion dichotomous fimbristylis herb of 345 weight portion Portugal of anemone rivularis, 315 weight portion little Hua orrisroots, 126 weight portion moth tree 114
107 weight portions.
3. a kind of pharmaceutical composition for preventing and treating rheumatoid arthritis according to claim 1, it is characterised in that drug regimen
Thing can prepare piece agent or capsule or dripping pill using the conventional method of galenic pharmacy.
4. a kind of pharmaceutical composition for preventing and treating rheumatoid arthritis according to claim 1, it is characterised in that drug regimen
The preventing and treating medicine for treating rheumatoid arthritis that thing is constituted with chemical drugs or Chinese medicine.
5. a kind of preparation method of the pharmaceutical composition for preventing and treating rheumatoid arthritis, it is characterised in that prepare as follows：
The composition and weight portion of bulk drug be：Anemone rivularis 342-348 weight portion Portugal 312-318 weight portion little Hua orrisroots
124-128 weight portion moth tree 112-116 weight portion dichotomous fimbristylis herb 105-109 weight portions；
Preparation method：
（1）Anemone rivularis, Portugal, little Hua orrisroots, moth tree, dichotomous fimbristylis herb are taken by bulk drug proportioning, is mixed, it is dense with percentage by weight
16% ethanol being spent as solvent, being taken in 37 DEG C of temperature extractions, extraction time is 18 times, and each extraction time is 65 hours, each solvent
Consumption is 44 times of bulk drug gross weight, is filtered, and obtains dregs of a decoction A and extract solution A, and extract solution A reclaims ethanol, is concentrated into relative density
1.08, filtration, liquid is first washed with water by SP70 large pore resin absorption columns, then with the ethanol solution of weight percent concentration 32%
Wash-out SP70 large pore resin absorption columns, collect the ethanol eluate of weight percent concentration 32%, reclaim ethanol, and concentrate drying is obtained final product
Extract A；
（2）Take step（1）Dregs of a decoction A, with the ethanol of weight percent concentration 48% as solvent, heating and refluxing extraction 31 times is carried every time
The time is taken for 0.3 hour, each solvent load is 78 times of dregs of a decoction A weight, and filtration obtains dregs of a decoction B and extract solution B, and extract solution B is returned
Ethanol is received, relative density 1.12 is concentrated into, filtered, liquid is first washed with water, then use by SIP1200 large pore resin absorption columns
The ethanol solution of weight percent concentration 86% elutes SIP1200 large pore resin absorption columns, collects the ethanol of weight percent concentration 86%
Eluent, reclaims ethanol, and concentrate drying obtains final product extract B；
（3）Extract A and extract B are mixed, pharmaceutical composition is obtained final product.
6. a kind of preparation method of the pharmaceutical composition for preventing and treating rheumatoid arthritis according to claim 5, its feature exists
Prepared in as follows：
The composition and weight portion of bulk drug be：The weight portion of 345 weight portion Portugal of anemone rivularis, 315 weight portion little Hua orrisroots 126
The weight portion of 114 weight portion dichotomous fimbristylis herb of moth tree 107；
Preparation method：
（1）Anemone rivularis, Portugal, little Hua orrisroots, moth tree, dichotomous fimbristylis herb are taken by bulk drug proportioning, is mixed, it is dense with percentage by weight
16% ethanol being spent as solvent, being taken in 37 DEG C of temperature extractions, extraction time is 18 times, and each extraction time is 65 hours, each solvent
Consumption is 44 times of bulk drug gross weight, is filtered, and obtains dregs of a decoction A and extract solution A, and extract solution A reclaims ethanol, is concentrated into relative density
1.08, filtration, liquid is first washed with water by SP70 large pore resin absorption columns, then with the ethanol solution of weight percent concentration 32%
Wash-out SP70 large pore resin absorption columns, collect the ethanol eluate of weight percent concentration 32%, reclaim ethanol, and concentrate drying is obtained final product
Extract A；
（2）Take step（1）Dregs of a decoction A, with the ethanol of weight percent concentration 48% as solvent, heating and refluxing extraction 31 times is carried every time
The time is taken for 0.3 hour, each solvent load is 78 times of dregs of a decoction A weight, and filtration obtains dregs of a decoction B and extract solution B, and extract solution B is returned
Ethanol is received, relative density 1.12 is concentrated into, filtered, liquid is first washed with water, then use by SIP1200 large pore resin absorption columns
The ethanol solution of weight percent concentration 86% elutes SIP1200 large pore resin absorption columns, collects the ethanol of weight percent concentration 86%
Eluent, reclaims ethanol, and concentrate drying obtains final product extract B；
（3）Extract A and extract B are mixed, pharmaceutical composition is obtained final product.
7. a kind of preparation method of the pharmaceutical composition for preventing and treating rheumatoid arthritis according to claim 5, its feature exists
Piece agent or capsule or dripping pill can be prepared using the conventional method of galenic pharmacy in pharmaceutical composition.
8. a kind of preparation method of the pharmaceutical composition for preventing and treating rheumatoid arthritis according to claim 5, its feature exists
In pharmaceutical composition and chemical drugs or Chinese medicine composition preventing and treating medicine for treating rheumatoid arthritis.
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