CN109223840A - A kind of pharmaceutical composition for treating knee osteoarthritis - Google Patents
A kind of pharmaceutical composition for treating knee osteoarthritis Download PDFInfo
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- CN109223840A CN109223840A CN201811492825.4A CN201811492825A CN109223840A CN 109223840 A CN109223840 A CN 109223840A CN 201811492825 A CN201811492825 A CN 201811492825A CN 109223840 A CN109223840 A CN 109223840A
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- achyranthis bidentatae
- radix achyranthis
- pharmaceutical composition
- knee osteoarthritis
- total saposins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
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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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
Abstract
The present invention relates to pharmaceutical technology field, the pharmaceutical composition of especially a kind for the treatment of knee osteoarthritis containing radix achyranthis bidentatae total saposins, gentiamarin and tanshinone IIA；The effective component of described pharmaceutical composition includes gentiamarin and radix achyranthis bidentatae total saposins；The present invention is on the basis of theory of traditional Chinese medical science, research Meng He doctor sends medication historical law, original new drug combination, compatibility is combined with more medicable Chinese medical extract, wherein gentiamarin is main therapeutic agent as monarch, radix achyranthis bidentatae total saposins containing 50% total saposins active component are minister, tanshinone IIA anti-inflammatory and antalgic is assistant, the theory of traditional Chinese medical science and saponin component ensuringd proper downward flow of the blood by radix achyranthis bidentatae have potential rush absorbent properties as surfactant, promote the therapeutic effect of gentiamarin and insoluble drug tanshinone IIA to knee osteoarthritis.
Description
Technical field
It is especially a kind of containing radix achyranthis bidentatae total saposins, gentiamarin and tanshinone IIA the present invention relates to pharmaceutical technology field
The pharmaceutical composition for treating knee osteoarthritis.
Background technique
The prosperity of the Schools TCMs researchs such as Meng He doctor group, and it is close to the attention of traditional Chinese culture and regression relation in recent years
It cuts.Chinese medicine needs to innovate, and with greater need for succession, innovation on the basis of succession just meets the rule of Development of Traditional Chinese Medicine and requires.Only
Innovation on the basis of traditional theory and method succession, is only the spy that the real Innovative TCMs compatibility of Chinese medicine is traditional Chinese medicine theory
Color and essence.Medicine is to being that the brief and exquisite experience that successive dynasties traditional Chinese medicine scholar accumulates during the side's of sending medication for a long time is total
Knot, although only two medicines are harmonious, arrange in pairs or groups that ingenious, ratio is proper, the characteristics of Chinese medicine " gregarious magical effect " can be annotated well.It excavates
Herbal pair in Schools TCM finds its excellent therapeutic effect, is the responsibility that modern traditional Chinese medicine worker should bear.
Knee osteoarthritis (Knee Osteoarthritis, KOA) is a kind of degenerative joint disease also known as knee joint hyperplasia
Property arthritis, degenerative osteoarthritis etc., using the regression of articular cartilage as major pathologic features, and involve synovium of joint, joint
The chronic inflammatory reaction of the periarticulars tissue such as capsule and periarticular osteophyte formation, menisic aging, joint space are narrow etc.
Pathological change seriously affects the life matter of patient so as to cause clinical symptoms such as knee joint swelling, pain, moving obstacle, deformities
Amount, is one of most common disease of the elderly.40 years old or less crowd is pointed out in the report of the famous emerging academician of orthopaedics house keeper Qiu Gui in China
Disease incidence about 5%, 60~75 years old crowd's disease incidence be higher than 5%, and crowd's disease incidence is up to 80% within 75 years old or more.The people of China six times
Mouth census data shows that 60 years old or more the elderly accounts for 13.26%, and with the rising of elderly population specific gravity, illness rate also rises therewith
Height brings heavy medical economy burden to individual, family, society, seriously affects the daily life and life of the middle-aged and the old
Quality.
For a long time, Meng He doctor group has always research and corresponding clinical treatment official documents and correspondence to record the treatment of rheumatism.?
In " four, Meng river doctor's collection ", Meng Heming cures Fei Boxiong for the single-row chapter of rheumatism, and illustrates the etiology and pathogenesis and warp of rheumatism
Proved recipe, it is indicated that " heresy of husband's six external factors which cause diseases, heat dryness-heat are sun, and chill is wet for yin.Negative gas repeatedly multiplies, obstructed circulation of nutrient QI and de fensive QI, passages through which vital energy circulates retardance, into bone and flesh
Three all diseases, and the disease of three numbness is made ", and point out that available three types of Arthralgia decoction, self-control temperature are supported flourish soup, Long Huotang, vertical pole soup etc. respectively
Treat wandering arthritis (migratory arthralgia), arthritis, numbness.Meng Heming doctor's horse training points out that " liver-kidney Yin deficiency, sun are bright wet in " arthroncus of knee " chapters and sections
Hot bettors, knee swell hot pain person, can't help heat consumption negative into pungent-warm " etc., it is believed that " so must assert fever and chills, with arteries and veins when diagnosis and treatment
Medication, this nothing are affected adversely ".
By investigation, it has been found that in the case that Meng river doctor treats bi Zheng, be by dredging collateral, support to seek and be controlled as common
Method.In treatment rheumatism medication, herbal pair includes " gentianae macrophyllae-radix achyranthis bidentatae " etc. for existing document report Meng He doctor group.Gentianae macrophyllae wind-damp dispelling,
Stopping numbness pain, clearind deficient heat；Radix achyranthis bidentatae dispelling stasis of blood and stimulating the menstrual flow, nourishing liver and kidney, strengthening the bones and muscles, inducing diuresis for treating strangurtia are ensuringd proper downward flow of the blood.Two medicines share, and gather dredging collateral altogether
Analgesic and feeding battalion's heat-clearing.
Gentianae macrophyllae is the dry root of gentianaceae plant gentianae macrophyllae (or gentiana straminea maxim, gentiana crassicaulis Duthie, radix gentiane dahuvicae).Its acrid flavour, hardship, property
It is flat.Return stomach, liver, gallbladder channel, there is wind-damp dispelling, stopping numbness pain, reducing the asthenic fever, clearing away damp-heat and other effects is conventionally used to rheumatic arthralgia, and tendon and vessel is arrested
Contraction, the diseases such as skeletal pain, modern pharmacological studies have shown that gentianae macrophyllae has the effects that anti-inflammatory, analgesia, calm, antipyretic, and related pharmacology
Effect has certain be associated with the iridoid glycosides such as gentiamarin etc. contained in gentianae macrophyllae.Wherein gentiamarin is iridoid
Class compound is easy to be hydrolyzed by gastric acid under one's belt, and due to its poorly water-soluble, it is organic to be generally dissolved in hot water or alcohols etc. after oral
Reagent, therefore be not easy to absorb.
Radix achyranthis bidentatae is the dry root of amaranthaceous plant radix achyranthis bidentatae, and bitter, acid are mild-natured.Return liver kidney channel, there is nourishing liver and kidney, strengthening the bones and muscles,
And " all medicine downlinks can be drawn " and other effects, it ranks first in the frequency of use for the treatment of bone and joint diseases.But the medicine of radix achyranthis bidentatae priming downlink
Imitate that material base and mechanism of action are unclear, existing research shows and the insectean metamorphosis activity sterone constituents that contain in radix achyranthis bidentatae
Such as ecdysterone, polysaccharide and triterpenoid saponins have certain association.Wherein the aglycon of saponin component has certain
Lipophilicity, sugar chain has stronger hydrophily again, saponins itself made to become a kind of surfactant.
Radix Salviae Miltiorrhizae is the dry root and rhizome of Lamiaceae plant, and bitter, slightly cold, converge to heart and liver channels have promoting blood circulation, stop
It bitterly, is the clinically used promoting blood circulation class drug of orthopedics and traumatology the effect of cool blood to disappear carbuncle.Existing research shows the anti-inflammatory effect master of Radix Salviae Miltiorrhizae
It is related with the tanshinone IIA contained in Radix Salviae Miltiorrhizae.Wherein tanshinone IIA is almost insoluble in water, influences the absorption of oral administration.
MMP-1, also known as clostridiopetidase A -1 are first by MMP that is qualitative and being purified from human fibroblasts, are had
Research shows that MMP-1 obviously increases in people's KOA synovial fluid.MMP-1 expresses most stable, cartilage degeneration during cartilage degeneration
More serious, MMP-1 expression is higher.
MMP-3, also known as stromelysins MMP-3 play critically important effect, previously studies have shown that KOA in KOA
MMP-3 content increases compared with Healthy People in patients serum and joint fluid, and MMP-3 is by cartilage cell, fibroblast, osteoblast
It secretes, MMP-3 expression is increased horizontal consistent with cartilage destruction degree in KOA articular cartilage synovia.
Summary of the invention
The object of the present invention is to provide a kind of, and the treatment kneecap containing radix achyranthis bidentatae total saposins, gentiamarin and tanshinone IIA closes
Scorching pharmaceutical composition is saved, the composition has the function of good anti-inflammatory and cartilage protection；By gentiamarin, gentianae macrophyllae total saposins
After tanshinone IIA compatibility, MMP-1 in knuckle synovia (p < 0.01) and MMP-3 (p < 0.05) can be significantly inhibited in cartilage cell
In expression, and stimulate the proliferation of chondrocytes, have the function of good anti-inflammatory and cartilage protection.
In order to solve the above technical problems, The technical solution adopted by the invention is as follows:
A kind of pharmaceutical composition for treating knee osteoarthritis, the effective component of described pharmaceutical composition include gentiamarin and
Radix achyranthis bidentatae total saposins.
Further, the mass ratio of the gentiamarin and radix achyranthis bidentatae total saposins is 1:5~5:1.
Further, the mass ratio of the gentiamarin and radix achyranthis bidentatae total saposins is 1:3.
It further, further include tanshinone IIA.
Further, the gentiamarin, radix achyranthis bidentatae total saposins, tanshinone IIA mass ratio are 1~5:1~5:1~5.
Further, the gentiamarin, radix achyranthis bidentatae total saposins, tanshinone IIA mass ratio are 1:3:1.
Further, which is characterized in that the radix achyranthis bidentatae total saposins the preparation method is as follows: radix achyranthis bidentatae through water or 0.1~30%
Ethyl alcohol extracts, and being concentrated into relative density is 1.02~1.10, is purified using D101 or HPD100 macroreticular resin, after purification
Radix achyranthis bidentatae total saponin content is no less than 50% in extract.
Further, described pharmaceutical composition is configured to oral preparation use.
Further, described pharmaceutical composition is for treating knee osteoarthritis illness.
It is using the beneficial effect of technical solution of the present invention:
Chinese medicine compatibility is assistant, attenuation to treat the drug of primary symptom and demonstrate,prove for monarch, enhancing or collaboration monarch drug in a prescription curative effect as minister, treatment time
Or coordinating the drug actions of a prescription is to make.For the present invention on the basis of theory of traditional Chinese medical science, research Meng He doctor sends medication historical law, original new drug combination side
Formula is combined compatibility with more medicable Chinese medical extract, and wherein gentiamarin is main therapeutic agent as monarch, contains
The radix achyranthis bidentatae total saposins of 50% total saposins active component are minister, tanshinone IIA anti-inflammatory and antalgic is assistant, are ensuringd proper downward flow of the blood by radix achyranthis bidentatae
Medical knowledge opinion and saponin component have as surfactant potentially promotees absorbent properties, promotes gentiamarin and insoluble drug red
Join therapeutic effect of the ketone IIA to knee osteoarthritis.
Specific embodiment
The following examples can make those skilled in the art that the present invention be more fully understood, but these embodiments are not
Limiting the scope of the invention." one embodiment " or " embodiment " referred to herein, which refers to, may be included in the present invention at least
A particular feature, structure, or characteristic in one implementation.In the present specification different places occur " in one embodiment
In " the same embodiment not is referred both to, nor the individual or selective embodiment mutually exclusive with other embodiments.
Embodiment 1
The embodiment of the invention provides a kind of for treating the pharmaceutical composition of knee osteoarthritis, pharmaceutical composition it is effective
Ingredient includes gentiamarin and radix achyranthis bidentatae total saposins, and the mass ratio of gentiamarin and radix achyranthis bidentatae total saposins is 1:5~5:1, and preferably 1:
3。
Embodiment 2
The embodiment of the invention provides a kind of for treating the pharmaceutical composition of knee osteoarthritis, pharmaceutical composition it is effective
Ingredient includes gentiamarin, radix achyranthis bidentatae total saposins and tanshinone IIA, and gentiamarin, radix achyranthis bidentatae total saposins, tanshinone IIA mass ratio are 1
~5:1~5:1~5, preferably gentiamarin, radix achyranthis bidentatae total saposins, tanshinone IIA mass ratio are 1:3:1.
Pharmaceutical composition in embodiment 1-2 is configured to oral preparation use, for treating knee osteoarthritis illness.
Radix achyranthis bidentatae total saposins in embodiment 1 and embodiment 2 the preparation method is as follows:
Method 1: taking radix achyranthis bidentatae medicine materical crude slice, adds water to cook twice, and every time plus 10 times of water, filtration, collecting decoction are concentrated into containing crude drug
0.5g/ml (relative density 1.02~1.05), upper D101 macroporous resin column are removal of impurities solvent with 5BV water, with 2BVh-1Flow velocity
Elution removal of impurities is carried out, water elution is discarded；Again with 50% ethyl alcohol 8BV, 4BVh-1Flow velocity is eluted, and eluent is collected.In order to
Concentration and dry efficiency are improved, reduces the loss of effective component, 70 DEG C or less are recovered under reduced pressure ethyl alcohol, are further concentrated to relative density
1.12~1.15 (70 DEG C), 70 DEG C or less vacuum drying, crush spare.Wherein radix achyranthis bidentatae total saponin content is no less than 50%.
Method 2: taking radix achyranthis bidentatae medicine materical crude slice, and 30% ethyl alcohol is added to extract twice, and every time plus 10 times of 30% ethyl alcohol of amount, filtration, merging mention
Liquid is taken, ethyl alcohol is recovered under reduced pressure, is concentrated into 0.5g/ml containing crude drug (relative density 1.02~1.05), upper D101 macroporous resin column, with
5BV water is removal of impurities solvent, carries out elution removal of impurities with 2BVh-1 flow velocity, discards water elution；Again with 60% ethyl alcohol 6BV, 4BV
H-1 flow velocity is eluted, and eluent is collected.In order to improve concentration and dry efficiency, reduce the loss of effective component, 70 DEG C with
Under ethyl alcohol is recovered under reduced pressure, be further concentrated to relative density 1.12~1.15 (70 DEG C), 70 DEG C or less vacuum drying, crush it is spare.Its
Middle radix achyranthis bidentatae total saponin content is no less than 60%.
One, KOA pharmacodynamics test is treated:
1, experimental material
(1) experimental animal
Take SD rat (half male and half female) 56,200 ± 20g of weight, 6 week old.
(2) major experimental instrument and instrument
Finland MK352 type microplate reader, SIGMA high-speed refrigerated centrifuge, Olympus inverted microscope, YD-6L organization embedding
Machine, RM2235 histotome, the cold dynamic platform of YD-6L organization embedding, TP1020 tissue processor, GNP9080 type water isolation type constant temperature
Incubator, Philips board electric shaver-for women, ultra low temperature freezer (Thermo scientific Forma 702), electronic scale is (often
State is double outstanding), board-washing machine etc..
(3) major experimental reagent
75% alcohol, 10% chloraldurate, Benzylpenicillin sodium salt, Yihong, dimethylbenzene, haematoxylin, goat serum, PBS buffer solution,
Primary antibody dilution (3%BSA in TBS, pH7.4), citrate buffer, DAB working solution etc., MMP-1, MMP-3 detection examination
Agent box (R&D systems company of the U.S.).
2, test method
(1) foundation of KOA model:
Anesthesia: taking SD rat, electronic scale weighing, and (0.3ml/100g) anesthesia is injected intraperitoneally in 10% chloraldurate.
Operation group: first buckling Rat Right knee, after alcohol disinfecting, it is seen that having a white ligament in front of knee joint is that kneecap is tough
Band cuts skin with blade on the inside of ligamentum patellae, appears ligamentum patellae, and knee-expanding position cuts inside ligamentum patellae, bends the knee kneecap is outside
Lateral dislocation sufficiently appears condyle of femur and shin bone, in femur and tibial medial gap, finds medial meniscus, cuts off inside half a month
1/3 before plate, then it is interrupted anterior cruciate ligament, stretching knee resets kneecap, and art mouth is given the drop prevention of 200,000 U/ml penicillin 2 and infected, and 1
Number silk suture art mouth.
Sham-operation group: cutting inside ligamentum patellae, after appearing articular cavity, does not damage meniscus and ligamentaum cruciatum, sutures joint
Capsule and skin.
All SD rats are postoperative to take right side gluteus maximus intramuscular injection penicillin prevention infection, every 0.3ml/ days, continuous 5
It.
(2) grouping administration: the SD rat of buying is grouped, respectively blank group, sham-operation group, Gentiana Water Extract group, ox
Knee decocting liquid group, gentiamarin-radix achyranthis bidentatae total saposins group, gentiamarin-radix achyranthis bidentatae total saposins-Tanshinone I IAGroup, diacerein (west
Medicine positive control drug) group, every group 8.Once a day.Successive administration 10 days, after administration in the 10th day after 2h, sacrificed by decapitation, and
Each tissue is taken out rapidly, washes away remained blood with physiological saline, it is spare in -80 DEG C of stored frozens.Testing index component content.
Radix achyranthis bidentatae decocting liquid configuration method: taking radix achyranthis bidentatae medicine materical crude slice, adds water to cook 2 times, and every time plus 8 glasss of water, decoction 1 hour filter,
Collecting decoction is concentrated into 1g/ml containing crude drug.
Gentiana Water Extract configuration method: taking gentianae macrophyllae medicine materical crude slice, adds water to cook 2 times, and every time plus 10 glasss of water, decoction 1 hour are filtered
It crosses, collecting decoction, is concentrated into 1g/ml containing crude drug.
Gentiamarin-radix achyranthis bidentatae total saposins group configuration method: gentiamarin and radix achyranthis bidentatae total saposins are weighed according to 1:3 ratio, is added
Appropriate hot water dissolving or dispersion are diluted to 1g/ml containing extract, can suitably shake up before or ultrasound makes to be uniformly dispersed.It is big every time
2ml, i.e. 2g extract is administered in mouse.
Gentiamarin-radix achyranthis bidentatae total saposins-tanshinone IIA group sample configuration method: rough gentian hardship is weighed according to 1:3:1 ratio
Glycosides, radix achyranthis bidentatae total saposins and tanshinone IIA add appropriate hot water dissolving or dispersion to be diluted to 1g/ml containing extract, can be appropriate before
It shakes up or ultrasound makes to be uniformly dispersed.2ml, i.e. 2g extract is administered in each rat.
(3) collection of specimens and preservation
Joint irrigation: rat after anesthesia being fixed, sterilizes art mouth, opens right side articular cavity, it is soft to visually observe knee joint
Bone and synovial membrane pump 0.5ml physiological saline repeated flushing articular cavity with 1ml syringe, take its joint fluid, be put into EP pipe, from
The heart (3000r/min) 5 minutes, takes supernatant, performs label, and -80 DEG C of stored frozens are spare.
Articular cartilage: after taking joint fluid, rat is put to death using cervical dislocation, right knee tibial plateau cartilage is taken, with 10%
Formaldehyde is fixed, decalcification, dehydration, transparent, paraffin embedding, and longitudinal 5 μm of serial section are stored at room temperature.
(4) degree of degeneration and synovial tissue's pathological change situation in knee cartilage face are visually observed.
(5) ELISA detects MMP-1, -3 contents in joint irrigation
Experimental principle: the tissue inhibiting using double antibody sandwich method measurement joint fluid MMP-1, -3 is horizontal.With purifying
MMPs antibody be coated with microwell plate, solid phase antibody is made, sequentially adds MMPs to single-phase antibody micropore is coated with, is marked with HRP
MMPs antibody combine, formed antibody-antigene-hrp-antibody complex, after washed, add substrate TMB develop the color；TMB is in HRP enzyme
Catalysis under convert au bleu, acid effect under be converted into yellow；MMPs in the depth and sample of color is positively correlated；Use enzyme
Mark instrument measures absorbance (OD value) at 450 nm, and the concentration of MMPs in sample is calculated by standard curve.
The dilution of standard items and sample-adding: being marked with quasi- 10 hole of sample wells on enzyme mark coating plate, the 1st, 2 holes add standard items μ l, point
Not Xi Shi after, each hole sample-adding amount is all 50 μ l, and concentration is respectively 150,100,50,25,12.5 μ l/l.
Sample-adding: setting blank well and sample to be tested hole respectively, is first loaded product dilution in sample to be tested hole on enzyme mark coating plate
Then 40 μ l of liquid adds 10 μ l of sample to be tested (the final dilution of sample is 5 times) again, sample is added on ELISA Plate hole bottom, as far as possible
Hole wall is not touched, mixing is shaked gently.
It incubates: with 37 DEG C of incubation 30min after sealing plate film sealing plate.
With liquid: will be spare after 20 times of concentrated cleaning solutions, 20 times of distilled water dilutions.
Washing: carefully taking sealing plate film off, discard liquid, dries, and cleaning solution is filled it up in every hole, discards after standing 30 seconds, so
It is repeated 5 times.
Enzyme: every hole is added 50 μ l of enzyme marking reagent, except blank well.
It incubates: 37 DEG C of incubation 30min.
Washing: removing liquid, dries, and cleaning solution is filled it up in every hole, discards after standing 30 seconds, is so repeated 5 times.
Colour developing: being added 50 μ l of color developing agent A, add 50 μ l of color developing agent B, and gently concussion mixes, and 37 DEG C are protected from light colour developing
15min。
Terminate: every hole adds 50 μ l of terminate liquid, terminates reaction.
Measurement: being returned to zero with blank well, and 450nm wavelength sequentially measures the OD value in each hole, establishes standard curve, and it is dense to calculate sample
Degree.
Statistical analysis: data are handled using SPSS21.0, two comparison among groups are examined using t, and more comparison among groups use variance
Analysis, inspection level α=0.05, p < 0.05 are statistically significant.
3, experimental result
(1) cartilage and synovial membrane regression situation are visually observed
Normal group and sham-operation group: articular cartilage surface is smooth, is in blue and white, flawless and softening stove, has no spur shape
At；The smooth no adhesion of synovial membrane；
Model group: articular cartilage forms in faint yellow, rough surface, local bony defect, periphery spur, has mucous membrane increasing
It is raw；
Gentianae macrophyllae group: articular cartilage is in faint yellow, surface is slightly coarse, local bony defect, periphery spur are formed, there have to be a small amount of viscous
Film hyperplasia；
Radix achyranthis bidentatae group: articular cartilage forms to yellow-white, rough surface, local bony defect, periphery spur, has in faint yellow
Mucosal hyperplasia；
Gentianae macrophyllae radix achyranthis bidentatae group: articular cartilage surface is essentially smooth, white to micro- red and white, there is a small amount of softening stove and spur shape
At synovial membrane is smooth substantially without adhesion；
Gentianae macrophyllae radix achyranthis bidentatae Radix Salviae Miltiorrhizae group: articular cartilage surface is essentially smooth, white to micro- red and white, there is a small amount of softening stove and bone
Superfluous to be formed, synovial membrane is smooth substantially without adhesion；
Diacerein group: articular cartilage surface is essentially smooth, white, has a small amount of softening stove and spur to be formed, synovial membrane light
It slides substantially without adhesion；
(2) knuckle synovia MMP-1, MMP-3 testing result
1 knuckle synovia MMP-1, MMP-3 testing result of table
Note: the comparision contents of MMP-1 and MMP-3 use one-way analysis of variance in each group knuckle synovia.* with sham-operation group
Compare, P < 0.05, * * is compared with sham-operation group, p < 0.01；Compared with model group, P < 0.054 discusses &
This results showed that
MMP-1 and MMP-3 is low in the expression of cartilage cell in sham-operation group, and MMP-1 and MMP-3 are in cartilage for model group
The expression of cell is significantly raised (p < 0.01).Illustrate that MMP-1 and MMP-3 can be used as grading articular cartilage in articular cartilage
The biological marker of regression.
In administration group, gentianae macrophyllae decocting liquid be applied alone that MMP-1 and MMP-3 can be inhibited in knuckle synovia in the table of cartilage cell
It reaches, but there was no significant difference compared with model group (p > 0.05).After gentiamarin and gentianae macrophyllae total saposins compatibility, can significantly it press down
Expression (p < 0.05) of the MMP-1 and MMP-3 in cartilage cell in knuckle synovia processed, and the proliferation of chondrocytes is stimulated, have good
The effect of good anti-inflammatory and cartilage protection.By gentiamarin, gentianae macrophyllae total saposins and Tanshinone I IAAfter compatibility, pass can be significantly inhibited
The expression of MMP-1 (p < 0.01) and MMP-3 (p < 0.05) in cartilage cell in synovia is saved, and stimulates the proliferation of chondrocytes, is had
There is good anti-inflammatory and cartilage protection.
Two, to the influence of intraperitoneal injection glacial acetic acid induced mice writhing response
Experiment purpose
Pharmaceutical composition is observed to the analgesic activity of mouse.
1 experimental material
1.1 test samples:
Radix achyranthis bidentatae decocting liquid configuration method: taking radix achyranthis bidentatae medicine materical crude slice, adds water to cook 2 times, and every time plus 8 glasss of water, decoction 1 hour filter,
Collecting decoction is concentrated into 1g/ml containing crude drug.
Gentiana Water Extract configuration method: taking gentianae macrophyllae medicine materical crude slice, adds water to cook 2 times, and every time plus 10 glasss of water, decoction 1 hour are filtered
It crosses, collecting decoction, is concentrated into 1g/ml containing crude drug.
Gentiamarin-radix achyranthis bidentatae total saposins group configuration method: gentiamarin and radix achyranthis bidentatae total saposins are weighed according to 1:3 ratio, is added
Appropriate hot water dissolving or dispersion are diluted to 0.5g/ml containing extract, can suitably shake up before or ultrasound makes to be uniformly dispersed.Every time
2ml, i.e. 1g extract is administered in rat.
Gentiamarin-radix achyranthis bidentatae total saposins-tanshinone IIA group sample configuration method: rough gentian hardship is weighed according to 1:3:1 ratio
Glycosides, radix achyranthis bidentatae total saposins and tanshinone IIA add appropriate hot water dissolving or dispersion to be diluted to 0.5g/ml containing extract, can fit before
When shaking up or ultrasound makes to be uniformly dispersed.2ml, i.e. 1g extract is administered in each rat.
1.2 reference substance
Panadol (paracetamol tablets), Sino-America Tianjin Shike Pharmaceutical Co., Ltd., 0.5g/ piece, 1 tablet once, and daily 3
It is secondary, i.e., 3 (1.5g) on the one.
Required concentration is according to dosage configured to distilled water when use, 4 DEG C of refrigerators save.
1.3 reagent
Glacial acetic acid, Guangzhou Chemical Reagent Factory.
1.4 instrument
JJ3000 animal electronic scale, G&G Products；BS224S electronic balance (1/0,000), German SARTORIUS product；
Mouse writhing scope, self-control.
1.5 animal
SPF grades of NIH mouse, half male and half female, 18~22g of weight.
2 experimental methods
2.1 animal packet
SPF grades of NIH mouse 72 are taken, half male and half female is randomly divided into 6 groups, respectively control group, panadol piece by weight
Group, Gentiana Water Extract group, radix achyranthis bidentatae decocting liquid group, gentiamarin-radix achyranthis bidentatae total saposins pharmaceutical composition group, gentiamarin-radix achyranthis bidentatae are total
Saponin(e-Tanshinone I IAPharmaceutical composition group.
The setting of 2.2 dosage
Panadol piece, according to package insert mark based on daily 1.5g clinic consumption per day, with reference to " herbal pharmacology is ground
Study carefully methodology " middle dosage conversion method " body surface area ratio " conversion animal clinical equivalent dose, using this clinical equivalent dosage as
Panadol piece mouse pharmacodynamic experiment dosage.Panadol piece mouse pharmacodynamic experiment dosage is 0.195gkg-1, to steam when experiment
Distilled water is according to dosage prepared, and 4 DEG C of refrigerators save.
Gentiana Water Extract, radix achyranthis bidentatae decocting liquid, gentiamarin-radix achyranthis bidentatae total saposins mixture, gentiamarin-radix achyranthis bidentatae total saposins-
Tanshinone I IA2ml is administered in mixture every time, is respectively equivalent to 2g crude drug or 1g extract.
2.3 experiment statistics methods
All measurement datas are plus or minus standard deviation from the meanIt indicates.The comparison of mean uses single factor test side between multiple groups
Difference analyses (One-Way ANOVA), and mean compares two-by-two between group, and SNK method is used when variance is neat；It is used when heterogeneity of variance
Dunnett ' s T3 method.It is completed by SPSSl5.0 software, α=0.05.
2.4 medication
Each dosage group of test sample drug and positive control drug according to dosage gastric infusion, administered volume 20mlkg-1, daily 1
Secondary, control group gives distilled water 20mlkg with method stomach-filling-1。
2.5 measuring method
SPF grades of NIH mouse 72 are taken, half male and half female is randomly divided into 6 groups, respectively control group, panadol piece by weight
Group, Jinan-Tai'an piece group, the prosperous prosperous high, medium and low dosage group of abstaining from drugs ball, every group 12.Each administration group according to dosage gastric infusion 20mL
kg-1, control group gives isometric distilled water with method stomach-filling, 1 time a day, continuous 5d.The equal fasting of groups of animals is not before last dose
Prohibit water 12h.1h after the last administration, every 0.7% glacial acetic acid 10mLkg of mouse ip-1, record the writhing incubation period of mouse in 20min
With writhing number, the difference of more each administration group and control group.
3 experimental results
Table 2 shows compared with the control group, gentiamarin-radix achyranthis bidentatae total saposins mixture group, gentiamarin-radix achyranthis bidentatae total saposins-
Tanshinone I IAMixture group mouse writhing incubation period is obviously prolonged (P < 0.01), writhing number substantially reduce (P < 0.01 or P <
0.05)。
Compared with panadol piece group, gentiamarin-radix achyranthis bidentatae total saposins mixture group, gentiamarin-radix achyranthis bidentatae total saposins-Radix Salviae Miltiorrhizae
Ketone IIAMixture group mouse writhing incubation period is obviously shortened (P < 0.05), and writhing number obviously increases (P < 0.05).
2 pharmaceutical composition of table to intraperitoneal injection glacial acetic acid induced mice writhing response influence (N=12)
Note: compared with the control group, P < 0.01 * P < 0.05, * *.Compared with panadol piece group, △ P < 0.05, △ △ P <
0.01。
Gentiamarin-radix achyranthis bidentatae total saposins medicinal mixture group and gentiamarin-radix achyranthis bidentatae total saposins-Tanshinone I IADrug is mixed
Closing object has obvious inhibiting effect to glacial acetic acid induced mice pain writhing response, illustrates that two kinds of pharmaceutical compositions have centainly
Analgesic activity.
Taking the above-mentioned ideal embodiment according to the present invention as inspiration, through the above description, relevant staff is complete
Various changes and amendments can be carried out without departing from the scope of the technological thought of the present invention' entirely.It is all in essence of the invention
Within mind and principle, any modification, equivalent substitution, improvement and etc. done be should all be included in the protection scope of the present invention.This
The technical scope of item invention is not limited to the contents of the specification, it is necessary to its technology is determined according to scope of the claims
Property range.
Claims (9)
1. a kind of pharmaceutical composition for treating knee osteoarthritis, it is characterised in that: the effective component of described pharmaceutical composition includes
Gentiamarin and radix achyranthis bidentatae total saposins.
2. a kind of pharmaceutical composition for treating knee osteoarthritis according to claim 1, it is characterised in that: the rough gentian is bitter
The mass ratio of glycosides and radix achyranthis bidentatae total saposins is 1:5~5:1.
3. a kind of pharmaceutical composition for treating knee osteoarthritis according to claim 2, it is characterised in that: the rough gentian is bitter
The mass ratio of glycosides and radix achyranthis bidentatae total saposins is 1:3.
4. a kind of pharmaceutical composition for treating knee osteoarthritis according to claim 1, it is characterised in that: further include Radix Salviae Miltiorrhizae
Ketone IIA.
5. a kind of pharmaceutical composition for treating knee osteoarthritis according to claim 4, it is characterised in that: the rough gentian is bitter
Glycosides, radix achyranthis bidentatae total saposins, tanshinone IIA mass ratio are 1~5:1~5:1~5.
6. a kind of pharmaceutical composition for treating knee osteoarthritis according to claim 5, it is characterised in that: the rough gentian is bitter
Glycosides, radix achyranthis bidentatae total saposins, tanshinone IIA mass ratio are 1:3:1.
7. a kind of pharmaceutical composition for treating knee osteoarthritis according to claim 1 to 6, which is characterized in that
The radix achyranthis bidentatae total saposins the preparation method is as follows: radix achyranthis bidentatae is extracted through water or 0.1~30% ethyl alcohol, being concentrated into relative density is 1.02
~1.10, it is purified using D101 or HPD100 macroreticular resin, radix achyranthis bidentatae total saponin content is no less than in extract after purification
50%。
8. a kind of pharmaceutical composition for treating knee osteoarthritis according to claim 7, it is characterised in that: the medicine group
It closes object and is configured to oral preparation use.
9. a kind of pharmaceutical composition for treating knee osteoarthritis according to claim 7, it is characterised in that: the medicine group
Object is closed for treating knee osteoarthritis illness.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201811492825.4A CN109223840A (en) | 2018-12-07 | 2018-12-07 | A kind of pharmaceutical composition for treating knee osteoarthritis |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811492825.4A CN109223840A (en) | 2018-12-07 | 2018-12-07 | A kind of pharmaceutical composition for treating knee osteoarthritis |
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Citations (3)
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| CN104127455A (en) * | 2014-07-18 | 2014-11-05 | 华侨大学 | Use of achyranthes root total saponins in drugs for preventing and treating osteoarthritis |
| CN104383301A (en) * | 2014-11-28 | 2015-03-04 | 山东省立医院 | Chinese patent medicine for treating osteoarthritis and preparation method thereof |
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