CN107080796A - A kind of Chinese medicine preparation for being used to treat Osteoarthritis - Google Patents
A kind of Chinese medicine preparation for being used to treat Osteoarthritis Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/32—Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/21—Amaranthaceae (Amaranth family), e.g. pigweed, rockwort or globe amaranth
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/59—Menispermaceae (Moonseed family), e.g. hyperbaena or coralbead
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/65—Paeoniaceae (Peony family), e.g. Chinese peony
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/894—Dioscoreaceae (Yam family)
- A61K36/8945—Dioscorea, e.g. yam, Chinese yam or water yam
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Abstract
The invention belongs to technical field of traditional Chinese medicines, pain caused by Osteoarthritis and hypertrophic arthritis, swelling, numbness, joint joint stuffiness, anchylosis, the Chinese medicine preparation of lopsided disease are particularly treated.The raw material medicaments in part by weight array of the Chinese medicine preparation turns into：8 13 parts of deer horn, 8 12 parts of the root of bidentate achyranthes, 8 12 parts of caulis sinomenii, 10 15 parts of rhizoma dioscoreae nipponicae, 8 13 parts of stir-baked RADIX PAEONIAE ALBA.The present invention is with caused by liver and kidney deficiency, and obstruction of meridians and collaterals is the interpretation of the cause, onset and process of an illness, based on classic classical prescription " deer horn ball ", Osteoarthritis clinical experience for many years is treated according to Affiliated Hospital of Liaoning University of Traditional Chinese Medicine orthopedics expert, drug dose is constantly adjusted, plus taste is formed, the clinical experience side summed up；It is pain caused by the treatment Osteoarthritis and hypertrophic arthritis being made up of pure Chinese medicine, swelling, numbness, joint joint stuffiness, anchylosis, the active drug of lopsided disease.Prove that there is significant analgesia, antiinflammatory action by zoopery.
Description
Technical field
The invention belongs to technical field of traditional Chinese medicines, more particularly to a kind of Chinese medicine preparation for being used to treat Osteoarthritis, especially
It is pain, swelling, numbness, joint joint stuffiness, anchylosis, deformity caused by treatment Osteoarthritis and hypertrophic arthritis
The Chinese medicine preparation of disease.
Background technology
Osteoarthritis is a kind of common disease, is mainly in after the middle age, category traditional Chinese medical science numbness disease-heumatism category.Numbness disease is by wind
The exopathogens such as cold-dampness attack human body, and numbness resistance channels and collaterals, QI-blood circulation is not smooth caused, is bent with the pain in joint, swelling, numbness, joint
Stretch it is unfavorable, very then anchylosis, deformity be main clinical manifestation.Liver governing tendons, Shen controls bone, woman six or seven, man six or eight, void declines it
As fading in, " irritability declines, and muscle is not active ", " kidney declines, body all pole " clinical visible muscular contracture and contraction, act it is forced, or impotence it is soft and
It is powerless.It is with the card of this disease actually asthenia in origin and asthenia in superficiality.This void, disease is in muscles and bones, and this is in caused by liver and kidney deficiency, and muscles and bones, which is lost, supports；Mark is real, and space between skin and muscles is empty
Void, wind-cold damp pathogen impatency channels and collaterals, sends out as numbness pain.State Administration of Traditional Chinese Medicine Eleventh Five-Year Plan heumatism (osteoarthritis) diagnosis and treatment scheme is faced
Bed checking is summarized in report, 320 patients, and syndrome of cold-dampness blocking collaterals is 53, and syndrome of dampness-heat blocking collaterals is 50, and caused by liver and kidney deficiency card is 196
Example, syndrome of deficiency of both qi and blood is 21, shows asthenia in origin and asthenia in superficiality, caused by liver and kidney deficiency is this sick main pathogenesis.Clinically Chinese traditional treatment bone
Arthritis is divided into internal therapy and external therapy, and wherein internal therapy is based on oral medicinal herb, and clinical common traditional Chinese medicine for oral administration prescription is more multiple
Miscellaneous, flavour of a drug quantity is more, and decocting method is more numerous, and cost of drugs is larger.Therefore, further research raw material it is few, it is with strong points in
Drug composition, it appears particularly important.
The content of the invention
In view of the above-mentioned problems, the present invention provides a kind of Chinese medicine preparation for being used to treat Osteoarthritis, bone is particularly treated
Property arthritis and hypertrophic arthritis caused by pain, swelling, numbness, joint joint stuffiness, anchylosis, lopsided disease.Should
Chinese medicine preparation raw material is few, curative effect is preferable, with strong points, clinical application effect is notable.
To achieve these goals, the present invention is adopted the following technical scheme that：The raw material medicaments in part by weight number of the Chinese medicine preparation
Constitute and be：8-13 parts of deer horn, 8-12 parts of the root of bidentate achyranthes, 8-12 parts of caulis sinomenii, 10-15 parts of rhizoma dioscoreae nipponicae, 8-13 parts of stir-baked RADIX PAEONIAE ALBA.
Described preparation is the clinically-acceptable pharmaceutical preparations such as pill, powder, mixture, disintegrated tablet or dispersible tablet.
The preparation method of the Chinese medicine preparation, comprises the following steps：Above-mentioned raw materials are matched by weight after weighing, plus
Decocting is boiled, and water is added by 8 times of medicine gross weight, is decocted 2 times, 1.5 hours every time, filtration, merging filtrate after concentration, was added
Formulation needed for clinically the acceptable auxiliary material of pharmaceutical preparation is made.
The property of medicine analysis of each raw material.
Deer horn：Salty, temperature, returns kidney, Liver Channel；With warm kidney sun, strengthening the bones and muscles, the effect of promoting circulation of blood is subsided a swelling.
Caulis sinomenii：It is bitter, pungent, flat, Return liver, the spleen channel；With wind-damp dispelling, the effect of degrading the channel.
The root of bidentate achyranthes：It is bitter, sweet, sour, flat, return liver and kidney channel；With filling liver kidney, strengthening the bones and muscles, the effect of dispelling stasis of blood and stimulating the menstrual flow.
Rhizoma dioscoreae nipponicae：Sweet, bitter, temperature, Return liver, kidney, lung channel；With dispelling wind and eliminating dampness, channels sootheing and network vessel quickening, the effect of promoting blood circulation and stopping pain.
The root of herbaceous peony：Acid, it is bitter, be slightly cold, Return liver, the spleen channel；With nourishing blood for regulating menstruation, the effect of soft liver relieves pain.
The prescription compatibility of the present invention clearly, complements each other between flavour of a drug；Wherein, the salty temperature Return liver kidney channel of deer horn taste, warm kidney sun,
Strengthening the bones and muscles, the effect of promoting circulation of blood detumescence, is monarch drug in a prescription.Root of bidentate achyranthes bitter acid flat return liver warp, with filling liver kidney, strengthening the bones and muscles, dispelling stasis of blood and stimulating the menstrual flow
Effect；Caulis sinomenii toil is put down, Return liver, the spleen channel, with wind-damp dispelling, the effect of degrading the channel；The two auxiliary deer horn treats the waist of heumatism
Knee joint soreness, muscles and bones is powerless, arthroncus, the disease such as limb paralysis, is for ministerial drug.Rhizoma dioscoreae nipponicae sweetness and bitterness temperature, Return liver, kidney, lung channel have
Dispelling wind and eliminating dampness, channels sootheing and network vessel quickening, the effect of promoting blood circulation and stopping pain；The root of herbaceous peony is bitter to be slightly cold, Return liver, the spleen channel, with nourishing blood for regulating menstruation, soft liver analgesic
Effect；The two auxiliary the medicine of the monarch and his subjects three, the arthroncus for treating heumatism, pain numbness, four limbs contraction pain etc. disease, be for assistant
Medicine.The root of bidentate achyranthes can ensuring proper downward flow of the blood, and to make medicine.All medicines share, can filling liver kidney, strengthening the bones and muscles, dispelling wind and eliminating dampness, promoting blood circulation and stopping pain plays a surname altogether
Numbness dredging collateral, the effect of dispelling stasis of blood and stimulating the menstrual flow.
Beneficial effects of the present invention.
The present invention is with caused by liver and kidney deficiency, and obstruction of meridians and collaterals is the interpretation of the cause, onset and process of an illness, based on classic classical prescription " deer horn " ball, according to Liaoning traditional Chinese medical science
Affiliated hospital of medicine university orthopedics expert treats Osteoarthritis clinical experience for many years, constantly adjusts drug dose, plus taste is formed, always
The clinical experience side born；It is pain, swelling, the fiber crops as caused by Osteoarthritis and hypertrophic arthritis being made up of pure Chinese medicine
The active drug of the disease such as wood, joint joint stuffiness, anchylosis, deformity；And prove have significantly by zoopery
Analgesia, antiinflammatory action.
Deer horn ball, come from Song Yanyong and《Prescriptions for Succouring the Sick》.The book is carried：" bone fictitious pole is controlled, edema of the face dirt is black, and rachiagia can not be stood long,
The deficiency of vital energy deals with that tooth is withered, and pain along spinal column very then likes saliva.Two liang of deer horn, the two halves of radix cyathulae one.Upper is smalls, such as making pellets by mixing medical powder with honey, Phoenix Tree Seed
Greatly, 70 balls are often taken, hollow salt soup is sent down ".
Deer horn filling liver kidney, strengthening the bones and muscles enter liver kidney channel are monarch drug in a prescription in we；Root of bidentate achyranthes tonifying liver and kidney and strengthening bones and muscles, dispelling stasis of blood and stimulating the menstrual flow, with green grass or young crops
Wind rattan wind-damp dispelling, degrading the channel is altogether ministerial drug, helps deer horn to treat the soreness of waist and knee joint of heumatism, and muscles and bones is powerless, arthroncus, limb paralysis
Deng disease.《Sheng Nong's herbal classic》Record the root of bidentate achyranthes：" bitter, acid, main cold-dampness phlegm numbness, spasm of limbs, goinyalgia are unable to flex and stretch the extremities, by vim and vigour, wound
It is showing tremendous enthusiasm rotten, induced abortion ".《Collected statements on the herbal foundation》Record：" caulis sinomenii, the medicine of dispersing wind and cold arthritis with fixed pain caused by dampness can relax the muscles and stimulate the blood circulation, the sharp marrow of bonesetting, therefore wind
Disease is weak and feeble, and the card that strength is attached undue importance to one thing to the neglect of the other by force, long term usage informal dress, makes outstanding contributions greatly "；Rhizoma dioscoreae nipponicae dispelling wind and eliminating dampness, channels sootheing and network vessel quickening, promoting blood circulation stops
Bitterly；Root of herbaceous peony nourishing blood for regulating menstruation, soft liver analgesic；The two auxiliary medicine of the monarch and his subjects three, the arthroncus for treating heumatism, pain numbness, four
The diseases such as limb contraction pain, are for adjutant.《Northeast medicine plants will》Record rhizoma dioscoreae nipponicae：" relax the muscles and stimulate the blood circulation, control lumbocrural pain, muscles and bones is numb "；《God
Agriculture book on Chinese herbal medicine warp》Call the root of herbaceous peony：" removing blood-arthralgia, break hard product, fever and chills hernia lump in the abdomen, analgesic ".The root of bidentate achyranthes can ensuring proper downward flow of the blood, and to make medicine.Five kinds of Chinese medicine
Share, catch the essence of this sick caused by liver and kidney deficiency, the work(for tonifying liver and kidney and strengthening bones and muscles are played altogether, removing obstruction in channels to relieve pain.Modern pharmacology shows, deer horn
With the destruction for reducing cartilage matrix, promote the effect of chondrocyte proliferation；The root of bidentate achyranthes plays important work in terms of anti-inflammatory, analgesia
With；Caulis sinomenii can suppress TNF-α and IL-1 β mRNA expressions；Rhizoma dioscoreae nipponicae can remove inflammatory cell, thin to mitigate synovial membrane
The inflammatory reaction of born of the same parents and disorganization；The root of herbaceous peony can be with pain of alleviation symptom.From Pharmacological Analysis, traditional Chinese medicine of the five flavours is mainly being adjusted simultaneously
Suppress to play anti-inflammatory, analgesic effect in terms of the generation of the related inflammatory factor.
Brief description of the drawings
The Chinese medicine preparation of Fig. 1 present invention is to anterior cruciate ligament detachment rat osteoarthritis HE coloration results.
The Chinese medicine preparation of Fig. 2 present invention is to anterior cruciate ligament detachment rat osteoarthritis Masson coloration results.
The Chinese medicine preparation of Fig. 3 present invention is to anterior cruciate ligament detachment rat osteoarthritis Toluidine blue staining result.
The Chinese medicine preparation of Fig. 4 present invention is to papain+cysteine induced rat osteoarthritis HE coloration results.
The Chinese medicine preparation of Fig. 5 present invention is tied to papain+cysteine induced rat osteoarthritis Masson dyeing
Really.
The Chinese medicine preparation of Fig. 6 present invention is to papain+cysteine induced rat osteoarthritis Toluidine blue staining
As a result.
Embodiment
With reference to specific embodiment, the present invention is further illustrated.
Embodiment 1.
The raw material medicaments in part by weight array of the herb mixture turns into：10 parts of deer horn, 12 parts of the root of bidentate achyranthes, 10 parts of caulis sinomenii wears mountain
15 parts of dragon, 10 parts of stir-baked RADIX PAEONIAE ALBA.
The preparation method of the herb mixture, comprises the following steps：Above-mentioned raw materials are matched by weight after weighing, plus
Decocting is boiled, and water is added by 8 times of medicine gross weight, is decocted 2 times, 1.5 hours every time, filtration, merging filtrate was concentrated into former mixing
/ 5th of object product, you can.
First, experimental method.
(1) Chinese medicine preparation of the present invention causes the arthritic protective effect of rat bone to study to anterior cruciate ligament detachment.
1st, packet and modeling.
180g~220g rats 112 are taken, male and female half and half, are randomly divided into 7 groups by every group 16, be respectively：(1) it is normal right
According to group：Distilled water 1ml/100g/d；(2) model group:Distilled water 1ml/100g/d；(3) the Chinese medicine preparation low dosage of embodiment 1
Group：5g crude drugs/kg/d；(4) the Chinese medicine preparation middle dose group of embodiment 1：15g crude drugs/kg/d；(5) Chinese medicine preparation of embodiment 1
High dose group：50g crude drugs/kg/d；(6) Chinese medicine positive controls：Zhuanggu guanjie pill, 1.2g/kg/d；(7) Western medicine positive control
Group：Diacerein, 0.01g/kg/d.
Modeling method：Rat dorsal position is fixed on plank, cuts off the leg in the knee joint cavity periphery 1cm regions to be injected
Hair, chelated iodine sterilization；Inner incision by right knee kneecap, successively opens articular cavity, kneecap is pushed away to being allowed to dislocate laterally, at knee joint
In maximum flexion state, exposure articular cavity cuts anterior cruciate ligament；Intersect before not cut after Normal group opening articular cavity tough
Band；The medicine given corresponding to above-mentioned every group is continuously given 8 weeks by each treatment group, gastric infusion, 2 times/day.
2nd, light Microscopic observation patellar surface damage degree scores.
24h after 8 weeks is administered, experimental animal all put to death by anesthesia, separates knee joint, exposure articular surface is observed, observed
The integrity degree and damaged condition of articular surface, standards of grading are as follows：0 point-articular surface finishing, color and luster is as usual；1 point-articular surface is coarse,
There is small crack and color and luster is gloomy；2 points-cartilage defect is as deep as cartilage middle level；3 points-articular surface ulcer is formed, cartilage defect as deep as
Cartilage deep layer；4 points-cartilage is stripped off, and subchondral bone matter is exposed；It the results are shown in Table 1.
Patellar surface damage degree scores under the light microscopic of table 1.
As a result show, the scoring of detachment model group is higher than Normal group, P<0.01, show the experimental model group modeling into
Work(, each dosage group of embodiment 1 is compared with detachment model group, statistically significant, P<0.01, and the high dose group of embodiment 1 with
Western medicine positive control drug diacerein is more statistically significant, P<0.05；It can be seen that, Chinese medicine preparation of the invention is tough to preceding intersection
Band detachment rat knee joints inflammation has protective effect.
3rd, Pathomorphology detection method.
Sample put into immediately after cutting 10% formaldehyde fix 48h, after enter 10% formic acid solution immersion, continuous decalcification 12d, extremely
Organize complete decalcification；Distilled water immersion, conventional dehydration, transparent, waxdip, embedding, 6 μm of section are saved backup, and carry out HE,
Masson, Toluidine blue staining, and do corresponding scoring.
3.1HE dyeing：Section is by the dewaxing of dimethylbenzene gradient, and haematine contaminates 5min, and washing reenters Yihong dip-dye
5min, washing, dimethylbenzene is transparent, neutral gum sealing.
As a result see Fig. 1, wherein 1- Normal groups, 2- detachments model group, 3- zhuanggu guanjie pills group, 4- diacereins group,
5- low dose groups, 6- middle dose groups, 7- high dose groups.
3.2Masson dyeing：Bounin fixers fix 48h, through 30% formic acid decalcification 48h, whole samples routinely stone
Wax section program processing, 7um thickness section, section moves to washing, enters PH6.0 Acetic acid-sodium acetate buffer solution dipping 30min
(37 DEG C), move into the hyaluronic acid enzymic digestion 6h that concentration is 0.5%, after washing, cut into slices and embathed into 2mol/L liquid calcium chlorides
48h, then Masson dyeing is done after washing, 10min is dyed in the mixing of Weigert haematoxylin A and Weigert haematoxylin B grades ratio,
Distillation washing 1min, Masson complex stainings liquid dyeing 15min, distilled water is slightly rinsed, 1% phosphomolybdic acid liquid processing 5min, aniline
Blue dyeing liquor redyes 5min, 1% glacial acetic acid water process 1min, is dehydrated transparent mounting.
As a result see Fig. 2, wherein 1- Normal groups, 2- detachments model group, 3- zhuanggu guanjie pills group, 4- diacereins group,
5- low dose groups, 6- middle dose groups, 7- high dose groups.
3.3 Toluidine blue staining：Toluidine blue dye liquor 30min, running water is rinsed, and glacial acetic acid breaks up clear to karyon, point
The change time is difficult long be otherwise easily caused and breaks up excessive, and running water is rinsed again, and dimethylbenzene is transparent after air-drying, and neutral gum is sealed
Gu.
As a result see Fig. 3, wherein 1- Normal groups, 2- detachments model group, 3- zhuanggu guanjie pills group, 4- diacereins group,
5- low dose groups, 6- middle dose groups, 7- high dose groups.
According to the dyeing of three of the above method, the scoring in terms of cartilage structure, quantity, matrix is carried out, standards of grading are such as
Under, it is shown in Table 2.
Standards of grading in terms of the cartilage structure of table 2, quantity, matrix.
Appraisal result such as table 3.
Table 3HE, Masson, Toluidine blue staining histological scores and comprehensive grading result.
Compared with normal group*P<0.05,**P<0.01, compared with detachment model group#P<0.05,##P<0.01, closed with zhuanggu
Section ball group compares Δ P<0.05,ΔΔP<0.01.
Three kinds of dyeing scorings of detachment model group and comprehensive grading are higher than Normal group, equal P<0.01, show the experiment
Model group modeling success, the comprehensive grading of each dosage group of embodiment 1 is compared with detachment group, statistically significant, equal P<0.01,
And low middle dose group is compared statistically significant with zhuanggu guanjie pill group, P<0.01.It can be seen that Chinese medicine preparation of the invention is handed over preceding
Pitching ligament detachment rat knee joints inflammation has protective effect.
4th, embodiment 1 to detachment anterior cruciate ligament rat blood serum IL-1 β, TNF-α, PGE2 contents influence.
It is administered 24h after 8 weeks, experimental animal is all anaesthetized, femoral artery takes blood, 3000 turns/min, 10min, take supernatant to carry out
Experiment, is operated, experimental result such as table 4 by IL-1 β, TNF-α, PGE2 kits explanation.
The rat blood serum IL-1 β of table 4, TNF-α, PGE2 content detection results.
| Group | Dosage (g/kg) | Number of elements | IL-1β(ng/L) | TNF-α(ng/L) | PGE2(ng/L) |
| Normal group | --- | 10 | 15.18±2.30 | 67.83±13.27 | 605.25±52.53 |
| Detachment group | --- | 10 | 19.72±3.68** | 92.45±19.23** | 668.90±71.71* |
| Low dose group | 5 | 10 | 14.27±3.17## | 81.49±17.67 | 595.02±59.18# |
| Middle dose group | 15 | 10 | 16.36±3.52## | 80.50±14.84 | 653.98±104.08 |
| High dose group | 50 | 10 | 15.78±2.48## | 91.31±16.02 | 648.78±80.22 |
| Zhuanggu guanjie pill group | 1.2 | 10 | 15.94±2.45## | 89.59±16.05 | 643.61±75.71 |
| Diacerein group | 0.01 | 10 | 15.07±2.77## | 92.28±24.02 | 574.63±67.47# |
As a result show, model group IL-1 β, TNF-α, PGE2 contents are compared with normal group raises, and show the experimental model
Group modeling success, embodiment 1 has reduction effect, P to the β of serum IL -1<0.01, it is not notable to TNF-α, PGE2 contents,
Only low dose group reduces serum PG E2 contents, there is significant difference, P<0.05.Compared * P with normal group<0.05, * * P<
0.01, Δ P is compared with model group<0.05, Δ Δ P<0.01.
(2) protective effect of Chinese medicine preparation of the invention to papain+cysteine induced rat osteoarthritis is ground
Study carefully.
1 packet and modeling.
180~220g rats 112 are taken, male and female half and half, are randomly divided into 7 groups by every group 16, be respectively：(1) normal control
Group：Distilled water 1ml/100g/d；(2) model group:Distilled water 1ml/100g/d；(3) low dose group of the Chinese medicine preparation of embodiment 1：
5g crude drugs/kg/d；(4) middle dose group of the Chinese medicine preparation of embodiment 1：15g crude drugs/kg/d；(5) height of the Chinese medicine preparation of embodiment 1
Dosage group：50g crude drugs/kg/d；(6) Chinese medicine positive controls：Zhuanggu guanjie pill, 1.2g/kg/d；(7) Western medicine positive controls：
Diacerein, 0.01g/kg/d.
Modeling method：The modeling liquid of 4% papain and 0.3M cysteines is prepared, dorsal position is fixed on plank,
Cut off the shank-feathering in the knee joint cavity periphery 1cm regions to be injected, chelated iodine sterilization.By 45 ° of rat knee joints flexing with kneecap
The Xiyan of lower extremely white kneecap tendon outer rim is entry point, punctures inserting needle to fossa intercondyloidea direction, arrives at and 2mm is withdrawn after condyle of femur, joint
Intracavitary slowly injects 0.1ml.Normal group injects isometric physiological saline.Each medication therapy groups continuously give said medicine
6 weeks, gastric infusion, 2 times/day.
2nd, light Microscopic observation patellar surface damage degree scores.
24h after 6 weeks is administered, experimental animal all put to death by anesthesia, separates knee joint, exposure articular surface is observed, observed
The integrity degree and damaged condition of articular surface, standards of grading are as follows：0 point-articular surface finishing, color and luster is as usual；1 point-articular surface is coarse,
There is small crack and color and luster is gloomy；2 points-cartilage defect is as deep as cartilage middle level；3 points-articular surface ulcer is formed, cartilage defect as deep as
Cartilage deep layer；4 points-cartilage is stripped off, and subchondral bone matter is exposed.It the results are shown in Table 5.
Patellar surface damage degree scores under the light microscopic of table 5.
As a result show, the scoring of papain+cysteine group is higher than Normal group, P<0.01, show the experiment
Model group modeling success, each dosage group of embodiment 1 is compared with papain+cysteine group, statistically significant, P<
0.01, Chinese medicine positive control drug zhuanggu guanjie pill group and Western medicine positive control drug diacerein and papain+cysteine
More statistically significant, the P of group<0.05.It can be seen that the present invention has to the rat knee joints inflammation of papain+cysteine induction
There is protective effect.Compared with normal group*P<0.05,**P<0.01, compared with papain+cysteine group#P<0.05,##P<
0.01。
3rd, Pathomorphology detection method.
Sample put into immediately after cutting 10% formaldehyde fix 48h, after enter 10% formic acid solution immersion, continuous decalcification 12d, extremely
Organize complete decalcification.Distilled water immersion, conventional dehydration, transparent, waxdip, embedding, 6 μm of section are saved backup, and carry out HE,
Masson, Toluidine blue staining, and do corresponding scoring.
3.1HE dyeing：Section is by the dewaxing of dimethylbenzene gradient, and haematine contaminates 5min, and washing reenters Yihong dip-dye
5min, washing, dimethylbenzene is transparent, neutral gum sealing.
As a result such as Fig. 4, wherein 1- Normal groups；2- papains+cysteine group；3- zhuanggu guanjie pill groups；4-
Diacerein group；5- low dose groups；6- middle dose groups；7- high dose groups.
3.2Masson dyeing：Bounin fixers fix 48h, through 30% formic acid decalcification 48h, whole samples routinely stone
Wax section program processing, 7um thickness section, section moves to washing, enters PH6.0 Acetic acid-sodium acetate buffer solution dipping 30min
(37 DEG C), move into the hyaluronic acid enzymic digestion 6h that concentration is 0.5%, after washing, cut into slices and embathed into 2mol/L liquid calcium chlorides
48h, then Masson dyeing is done after washing, 10min is dyed in the mixing of Weigert haematoxylin A and Weigert haematoxylin B grades ratio,
Distillation washing 1min, Masson complex stainings liquid dyeing 15min, distilled water is slightly rinsed, 1% phosphomolybdic acid liquid processing 5min, aniline
Blue dyeing liquor redyes 5min, 1% glacial acetic acid water process 1min, is dehydrated transparent mounting.
As a result such as Fig. 5, wherein 1- Normal groups；2- papains+cysteine group；3- zhuanggu guanjie pill groups；4-
Diacerein group；5- low dose groups；6- middle dose groups；7- high dose groups.
3.3 Toluidine blue staining：Toluidine blue dye liquor 30min, running water is rinsed, and glacial acetic acid breaks up clear to karyon, point
The change time is difficult long be otherwise easily caused and breaks up excessive, and running water is rinsed again, and dimethylbenzene is transparent after air-drying, and neutral gum is sealed
Gu.
As a result such as Fig. 6, wherein 1- Normal groups；2- papains+cysteine group；3- zhuanggu guanjie pill groups；4-
Diacerein group；5- low dose groups；6- middle dose groups；7- high dose groups.
According to the dyeing of three of the above method, the scoring in terms of cartilage structure, quantity, matrix is carried out, standards of grading are such as
Table 6.
Standards of grading in terms of the cartilage structure of table 6, quantity, matrix.
Appraisal result is shown in Table 7.
Table 7HE, Masson, Toluidine blue staining histological scores and comprehensive grading result.
Compared with normal group*P<0.05,**P<0.01, compared with model group#P<0.05,##P<0.01。
Three kinds of dyeing scorings of papain+cysteine group and comprehensive grading are higher than Normal group, equal P<0.01,
Show experimental model group modeling success, toluidine blue and comprehensive grading and the Guang of papain+half of each dosage group of embodiment 1
Propylhomoserin group compares, statistically significant, equal P<0.01, the HE of middle high dose group and papain+cysteine group is dyed
Score more statistically significant, P<0.01.Chinese and Western positive drug is in each dyeing scoring and papain+cysteine group ratio
It is more statistically significant, P<0.05, P<0.01, it is seen that Chinese medicine preparation of the invention is big to papain+cysteine group
Mouse gonitis has protective effect.
4. embodiment 1 induces gonitis rat blood serum IL-1 β, TNF-α, PGE2 to contain papain+cysteine
The influence of amount.
It is administered 24h after 6 weeks, experimental animal is all anaesthetized, femoral artery takes blood, 3000 turns/min, 10min, take supernatant to carry out
Experiment, is operated, experimental result such as table 8 by IL-1 β, TNF-α, PGE2 kits explanation.
The rat blood serum IL-1 β of table 8, TNF-α, PGE2 content detection results.
As a result show, papain+cysteine group IL-1 β, TNF-α content is compared with normal group raises, show
Experimental model group modeling success, middle dose group has reduction effect, P to the β of serum IL -1<0.01, to TNF-α, PGE2
Content is not notable, and middle dose group is compared with papain+cysteine group, the reduction of the β of serum IL -1 contents, P<0.05, middle dose
Amount group is compared with papain+cysteine group, the reduction of the β of serum IL -1 contents, P<0.05, high dose group and papain
+ cysteine group compares, the reduction of TNF-α content, P<0.01, and compared with zhuanggu guanjie pill group, TNF-α content drop
It is low, P<0.05.Compared * P with normal group<0.05,**P<0.01, compared with model group#P<0.05,##P<0.01, with zhuanggu joint
Ball group compares Δ P<0.05,ΔΔP<0.01.
Embodiment 2.
The raw material medicaments in part by weight array of the Chinese medicine preparation turns into：8 parts of deer horn, 8 parts of the root of bidentate achyranthes, 12 parts of caulis sinomenii, rhizoma dioscoreae nipponicae
13 parts, 8 parts of stir-baked RADIX PAEONIAE ALBA.
Embodiment 3.
The raw material medicaments in part by weight array of the Chinese medicine preparation turns into：13 parts of deer horn, 10 parts of the root of bidentate achyranthes, 8 parts of caulis sinomenii, rhizoma dioscoreae nipponicae
10 parts, 13 parts of stir-baked RADIX PAEONIAE ALBA.
2nd, model case.
Case 1：It is remaining certain, man, 56 years old；Main suit：Left gonalgia 1 year, aggravates two weeks.It is existing symptoms include：Left knee joint pain
Bitterly, pain is in shouting pain, and fixed pain, dark tongue quality, tongue is white, veins string；Without past medical history；Without allergies；Training is checked：Left knee swelling
(-), interior lateral articular gap tenderness (+), limitation of activity (+), floating patella test (-), McMurray sign (-), front and rear drawer test and side
To stress test (-)；Imageological examination：Left knee joint anterioposterior and lateral film shows that inner side joint space is narrow, constitutes the different degrees of bone of bone
Matter hyperplasia；Tcm diagnosis：Rheumatism (syndrome of blood stasis and stagnant qi), doctor trained in Western medicine diagnosis：Left knee osteoarthritis；Therapy and formula：The system of embodiment 1
14 doses of agent, twice daily orally, medication rear left knee pain symptom are clearly better.
Case 2：Zhang, female, 63 years old；Main suit：Right knee joint swelling pain 3 months, aggravates companion's anchylosis 4 days；Existing disease
See：Right knee joint swelling pain, soreness and weakness of waist and knees, red tongue body, tongue is white, deep thready pulse；Past medical history：Diabetes medical history 5 years, takes diformazan certainly
Biguanides sustained release tablets, glycemic control is stable；Without allergies；Training is checked：Right knee swelling (+), lateral articular gap tenderness (+) is living
Dynamic limited (++), floating patella test (-), McMurray sign (-), front and rear drawer test and lateral stress experiment (-), the right apodization function of foot five are lived
Dynamic and dermal sensation is normal；Tcm diagnosis：Rheumatism quench one's thirst (caused by liver and kidney deficiency card) doctor trained in Western medicine diagnosis：The right type glycosuria of knee osteoarthritis 2
Disease；Therapy and formula：14 doses of the preparation of embodiment 2, day is oral twice, and medication rear right knee pain symptom mitigates, and 14 doses, symptom are taken again
It is clearly better earlier above.
Case 3：Liu, man, 59 years old；Main suit：Double knee joint pain 6 months, is aggravated 2 days；It is existing symptoms include：Double knee joint aches
Bitterly, pain is in shouting pain, and fixed pain, day at light night is heavy, and dark tongue quality tongue is white, veins string；Without past medical history；Without allergies；Training is checked：It is double
Joint space tenderness (+) on the inside of knee swelling (-), left knee lateral articular gap tenderness (+), right knee, limitation of activity (+) floats kneecap examination
Test (-), McMurray sign (-), front and rear drawer test and lateral stress experiment (-), the activity of the apodization function of biped five and dermal sensation are normal,
Peripheral blood is law-abiding.Imageological examination：Double knee joint anterioposterior and lateral film shows that intercondylar ridge comes to a point on the inside of left knee, and kneecap arnold ligament ossify；Right knee
Joint lateral gap narrows, and constitutes the different degrees of osteoproliferation of bone；Tcm diagnosis：Rheumatism (syndrome of blood stasis and stagnant qi), doctor trained in Western medicine diagnosis：It is double
Knee osteoarthritis therapy and formula：20 doses of the preparation of embodiment 3, day is oral twice.Rear left knee pain symptom of taking medicine mitigates, right knee
Improve unobvious, 15 doses are taken again, right knee pain symptom is clearly better earlier above.
Case 4：Yan, female, 53 years old, main suit：Flank pain, limitation of activity 3 years is aggravated 10 days.It is existing symptoms include：Waist aches
Tong , Acid are soft, and leg knee is powerless, meet labor and aggravate, recurrent exerbation, pale tongue, deep thready pulse.Without past medical history；Without allergies；Training is checked：Waist
Portion's physiological camber is straightened, and tenderness (+) at 1.5cm is opened on the spinous process of waist 4- sacrums 1 and side, and left vertical leg raises 50 ° (+) of experiment, right straight legs lift
70 ° (-) of height experiment, strengthens experiment (-), sciatic stretch test (-), " 4 " word experiment (-), babinski sign (-), double lower limb flesh
Power, dermal sensation are normally and the activity of five apodization functions is normal.Imageological examination：Waist CT shows waist 4-5, the disc herniation of waist 5- sacrums 1.
Tcm diagnosis：Lumbago diseases (caused by liver and kidney deficiency card), doctor trained in Western medicine diagnosis：The protrasion of the lumbar intervertebral disci.Therapy and formula：The preparation 14 of embodiment 1
Agent, day is oral twice.Symptom after taking medicine mitigates, and 14 doses is taken again, symptom is clearly better.
Claims (6)
1. a kind of be used to treat the Chinese medicine preparation of Osteoarthritis, it is characterised in that its raw material medicaments in part by weight array into including：
8-13 parts of deer horn, 8-12 parts of the root of bidentate achyranthes, 8-12 parts of caulis sinomenii, 10-15 parts of rhizoma dioscoreae nipponicae, 8-13 parts of stir-baked RADIX PAEONIAE ALBA.
2. the Chinese medicine preparation as claimed in claim 1 for being used to treat Osteoarthritis, it is characterised in that its bulk drug presses number
Constitute and be：10 parts of deer horn, 12 parts of the root of bidentate achyranthes, 10 parts of caulis sinomenii, 15 parts of rhizoma dioscoreae nipponicae, 10 parts of stir-baked RADIX PAEONIAE ALBA.
3. the Chinese medicine preparation as claimed in claim 1 for being used to treat Osteoarthritis, it is characterised in that its bulk drug is by weight
Number is constituted：8 parts of deer horn, 8 parts of the root of bidentate achyranthes, 12 parts of caulis sinomenii, 13 parts of rhizoma dioscoreae nipponicae, 8 parts of stir-baked RADIX PAEONIAE ALBA.
4. the Chinese medicine preparation as claimed in claim 1 for being used to treat Osteoarthritis, it is characterised in that its bulk drug is by weight
Number is constituted：13 parts of deer horn, 10 parts of the root of bidentate achyranthes, 8 parts of Sabia japonica, 10 parts of rhizoma dioscoreae nipponicae, 13 parts of stir-baked RADIX PAEONIAE ALBA.
5. being used for as described in claim 1-4 is any treats the Chinese medicine preparation of Osteoarthritis, it is characterised in that described system
Agent is the clinically-acceptable pharmaceutical preparations such as pill, powder, mixture, disintegrated tablet or dispersible tablet.
6. being used for as described in claim 1-5 is any treats the preparation method of the Chinese medicine preparation of Osteoarthritis, its feature exists
In above-mentioned raw materials being matched by weight after weighing, added water to cook, water are added by 8 times of medicine gross mass, decoct 2 times, often
Secondary 1.5 hours, filtration, merging filtrate, concentration, you can.
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| CN117045733A (en) * | 2023-09-26 | 2023-11-14 | 辽宁中医药大学 | Extraction and purification method of traditional Chinese medicine compound for treating knee osteoarthritis |
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