Four, the specific embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is elaborated.
Embodiment 1
The present invention in the specific implementation, be to make: Radix Angelicae Sinensis 90g by following medicine, Rhizoma Chuanxiong 100g, Pericarpium Zanthoxyli 100g, Rhizoma Et Radix Notopterygii 90g, Rhizoma Polygoni Cuspidati 90g, Herba Asari 90g, Radix Angelicae Dahuricae 90g, Radix Aconiti Preparata 90g, Herba Sarcandrae 90g, Sanguis Draxonis 25g, Borneolum Syntheticum 18g and Mentholum 18g, wherein, earlier with Radix Angelicae Sinensis, Rhizoma Chuanxiong, Pericarpium Zanthoxyli, Rhizoma Et Radix Notopterygii, Rhizoma Polygoni Cuspidati, Herba Asari, the Radix Angelicae Dahuricae, Radix Aconiti Preparata and Herba Sarcandrae nine flavor Chinese medicines mix, be ground into coarse powder, according to the percolation under the extractum item, use Radix Angelicae Sinensis, Rhizoma Chuanxiong, Pericarpium Zanthoxyli, Rhizoma Et Radix Notopterygii, Rhizoma Polygoni Cuspidati, Herba Asari, the Radix Angelicae Dahuricae, Radix Aconiti Preparata and Herba Sarcandrae weight and 8 times of amount mass concentrations be 70% ethanol, speed percolation with 5ml/min, collect percolate, reclaim ethanol, standby; Sanguis Draxonis is with alcohol dipping twice, and the mass concentration that at every turn adds 10 times of Sanguis Draxonis weight is 70% ethanol, floods 24 hours, merges impregnation liquid, and is standby; Borneolum Syntheticum and Mentholum are with the dissolve with ethanol of mass concentration 30%, and be standby; Merge above-mentioned three kinds of medicinal liquids, adjust and to contain the alcohol amount to reach mass concentration be 30%, leave standstill filtration, getting filtrate, to add mass concentration be that 30% ethanol is adjusted to every milliliter of medicinal liquid and contains and be equivalent to 1 gram crude drug, promptly gets the Chinese medicine pain relieving spray of the treatment osteoarthrosis pain of 1000ml.
Embodiment 2
The present invention is in concrete enforcement, also can be by Radix Angelicae Sinensis 100g, Rhizoma Chuanxiong 120g, Pericarpium Zanthoxyli 120g, Rhizoma Et Radix Notopterygii 100g, Rhizoma Polygoni Cuspidati 100g, Herba Asari 100g, Radix Angelicae Dahuricae 100g, Radix Aconiti Preparata 100g, Herba Sarcandrae 100g, Sanguis Draxonis 30g, Borneolum Syntheticum 20g, Mentholum 20g makes, wherein, earlier with Radix Angelicae Sinensis, Rhizoma Chuanxiong, Pericarpium Zanthoxyli, Rhizoma Et Radix Notopterygii, Rhizoma Polygoni Cuspidati, Herba Asari, the Radix Angelicae Dahuricae, Radix Aconiti Preparata and Herba Sarcandrae nine flavor Chinese medicines mix, be ground into coarse powder, according to the percolation under the extractum item, use Radix Angelicae Sinensis, Rhizoma Chuanxiong, Pericarpium Zanthoxyli, Rhizoma Et Radix Notopterygii, Rhizoma Polygoni Cuspidati, Herba Asari, the Radix Angelicae Dahuricae, Radix Aconiti Preparata and Herba Sarcandrae weight and 8 times of amount mass concentrations be 70% ethanol, speed percolation with 5ml/min, collect percolate, reclaim ethanol, standby; Sanguis Draxonis is with alcohol dipping twice, and the mass concentration that at every turn adds 10 times of Sanguis Draxonis weight is 70% ethanol, floods 24 hours, merges impregnation liquid, and is standby; Borneolum Syntheticum and Mentholum are with the dissolve with ethanol of mass concentration 30%, and be standby; Merge above-mentioned three kinds of medicinal liquids, adjust and to contain the alcohol amount to reach mass concentration be 30%, leave standstill filtration, getting filtrate, to add mass concentration be that 30% ethanol is adjusted to every milliliter of medicinal liquid and contains and be equivalent to 1 gram crude drug, promptly gets the Chinese medicine pain relieving spray of the treatment osteoarthrosis pain of 1000ml.
Embodiment 3
The present invention is when implementing, also can be by Radix Angelicae Sinensis 110g, Rhizoma Chuanxiong 140g, Pericarpium Zanthoxyli 140g, Rhizoma Et Radix Notopterygii 110g, Rhizoma Polygoni Cuspidati 110g, Herba Asari 110g, Radix Angelicae Dahuricae 110g, Radix Aconiti Preparata 110g, Herba Sarcandrae 110g, Sanguis Draxonis 35g, Borneolum Syntheticum 22g, Mentholum 22g makes, wherein, earlier with Radix Angelicae Sinensis, Rhizoma Chuanxiong, Pericarpium Zanthoxyli, Rhizoma Et Radix Notopterygii, Rhizoma Polygoni Cuspidati, Herba Asari, the Radix Angelicae Dahuricae, Radix Aconiti Preparata and Herba Sarcandrae nine flavor Chinese medicines mix, be ground into coarse powder, according to the percolation under the extractum item, use Radix Angelicae Sinensis, Rhizoma Chuanxiong, Pericarpium Zanthoxyli, Rhizoma Et Radix Notopterygii, Rhizoma Polygoni Cuspidati, Herba Asari, the Radix Angelicae Dahuricae, Radix Aconiti Preparata and Herba Sarcandrae weight and 8 times of amount mass concentrations be 70% ethanol, speed percolation with 5ml/min is collected percolate, reclaim ethanol, standby; Sanguis Draxonis is with alcohol dipping twice, and the mass concentration that at every turn adds 10 times of Sanguis Draxonis weight is 70% ethanol, floods 24 hours, merges impregnation liquid, and is standby; Borneolum Syntheticum and Mentholum are with the dissolve with ethanol of mass concentration 30%, and be standby; Merge above-mentioned three kinds of medicinal liquids, adjust and to contain the alcohol amount to reach mass concentration be 30%, leave standstill filtration, getting filtrate, to add mass concentration be that 30% ethanol is adjusted to every milliliter of medicinal liquid and contains and be equivalent to 1 gram crude drug, promptly gets the Chinese medicine pain relieving spray of the treatment osteoarthrosis pain of 1000ml.
The science compatibility of said medicine, and through science preparation, support to have damp eliminating cold expelling, relaxing muscles and tendons and activating QI and blood in the collateral, blood circulation promoting and blood stasis dispelling, reducing swelling and alleviating pain mutually has the good curing effect to bone joint pain etc., and has obtained fully proving that interrelated data is as follows through experiment:
1, reagent and instrument
1.1 reagent: spray of the present invention is an experiment product, and aconitine is a reference substance, and when being classified as control medicinal material, Rhizoma Chuanxiong control medicinal material, above reference substance be all available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, silica gel G (Qingdao Haiyang chemical industry group), and agents useful for same is analytical pure.
1.2 instrument: electronic balance (Sartorius); Quantitative capillary tube (Drummond co.); Three usefulness ultraviolet analyzers (Shanghai ZF-6 type)
2. Study on Preparation of the present invention, experiment green pepper rhizome of chuanxiong spray are to be made by prescription of the present invention by Radix Angelicae Sinensis, Rhizoma Chuanxiong, Pericarpium Zanthoxyli, Rhizoma Et Radix Notopterygii, Rhizoma Polygoni Cuspidati, Herba Asari, the Radix Angelicae Dahuricae, Radix Aconiti Preparata, Herba Sarcandrae, Sanguis Draxonis, Borneolum Syntheticum and Mentholum, physicochemical property according to square Chinese crude drug pharmacological experiments and contained effective ingredient or effective site, Rhizoma Chuanxiong, Radix Angelicae Sinensis, Pericarpium Zanthoxyli, Rhizoma Et Radix Notopterygii, Rhizoma Polygoni Cuspidati, Herba Asari, the Radix Angelicae Dahuricae, Radix Aconiti Preparata and Herba Sarcandrae nine flavor Chinese medicines mix in will writing out a prescription, be ground into coarse powder, use ethanol percolation; With the Sanguis Draxonis alcohol dipping, extract its effective ingredient; Borneolum Syntheticum and Mentholum is standby with 30% dissolve with ethanol.
2.1 extraction process is investigated
2.1.1 percolation technology is investigated according to the preparation technology's factor that influence this product quality: the design that experimentizes of determining alcohol (A), ethanol consumption (B), the following factor level table of percolation speed (C) selection sees Table 1
Table 1 percolation factor level table
Take by weighing Rhizoma Chuanxiong by prescription, Radix Angelicae Sinensis etc., totally 9 parts, carry out percolation by each experimental program in table 1 factor level and the table 2 (), extracting solution reclaims ethanol, is concentrated into volume and is 100ml, and is standby.The accurate 10ml that draws puts in the evaporating dish that is dried to constant weight, water bath method, and dry 3 hours of 105 degree were put in the exsiccator 30 minutes, weighed immediately.
Extractum leaching amount is carried out intuitive analysis, and optimum extraction condition is 8 times of amounts of 70% ethanol, carries out percolation with 5ml/min speed.
2.1.2 impregnation technology is investigated the character according to chemical constituent in the Sanguis Draxonis, in conjunction with the character of impregnation liquid, we are solvent impregnated with 70% ethanol, ethanol, and relatively the character of lixiviating solution is decided to be at last with alcohol dipping twice, adds 10 times of amounts of ethanol at every turn, floods 24 hours.
2.2 preparation technology
2.2.1 the definite of extraction process gets Radix Angelicae Sinensis, Rhizoma Chuanxiong, Pericarpium Zanthoxyli, Rhizoma Et Radix Notopterygii, Rhizoma Polygoni Cuspidati, Herba Asari, the Radix Angelicae Dahuricae, Radix Aconiti Preparata and Herba Sarcandrae nine flavor Chinese medicines in the prescription ratio, be ground into coarse powder, according to the percolation under the extractum item (" appendix IO of Chinese pharmacopoeia version in 2000), with 8 times of amount 70% ethanol, speed percolation with 5ml/min, collect percolate, reclaim ethanol, standby.Sanguis Draxonis adds 10 times of amounts of ethanol with alcohol dipping twice at every turn, floods 24 hours, and it is standby to merge impregnation liquid.Borneolum Syntheticum and Mentholum are standby with 30% dissolve with ethanol.
2.2.2 the above-mentioned three kinds of medicinal liquids of definite merging of dosing technology are adjusted and to be contained the alcohol amount and reach 30%, leave standstill filtration, get filtrate and add 30% ethanol and be adjusted to every milliliter and be equivalent to 1 gram crude drug promptly.Above-mentioned recipe quantity prepares finished product 1000ml.
3. quality standard research
3.1 differentiating, the thin layer of Radix Angelicae Sinensis, Rhizoma Chuanxiong gets this product 10ml, water bath method, and adding distil water 10ml dissolving adds n-hexane extraction twice, each 20ml, combining extraction liquid, water-bath concentrates, and is concentrated into 1ml, as test sample liquid.Get Rhizoma Chuanxiong control medicinal material 1g, Radix Angelicae Sinensis control medicinal material 1 gram adds normal hexane 20ml respectively, shines medical material solution in pairs with legal system.Make scarce Radix Angelicae Sinensis, Rhizoma Chuanxiong sample in proportion in addition, make negative controls by the preparation method of test sample liquid.According to the thin layer chromatography test, draw each 2 μ l of above-mentioned four kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with normal hexane-ethyl acetate (9: 1), launch, take out, dry, put to examine under the ultra-violet lamp (365nm) and know.In the test sample chromatograph, with control medicinal material chromatograph relevant position on, show three identical fluorescence speckles, the negative control liquid chromatography does not then have this speckle.
Get this product 20ml 3.2 the thin layer of Borneolum Syntheticum is differentiated, water-bath is steamed to there not being the alcohol flavor, adds chloroform 30ml extracting twice, combining extraction liquid, and water bath method (low temperature), residue add chloroform 1ml makes dissolving, as need testing solution.Get the negative control product 20ml that lacks Borneolum Syntheticum, make negative control product solution with method.Other gets the Borneolum Syntheticum reference substance, and chlorination is copied into the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix V of Chinese Pharmacopoeia version in 2000 I B), draw need testing solution 2 μ l, draw negative control product solution 2 μ l, draw reference substance solution 10 μ l, put respectively on same silica gel g thin-layer plate, with benzene-ethyl acetate (17: 1) is developing solvent, launch, take out, dry, spray is with 5% phosphomolybdic acid ethanol solution, about 10 minutes of 105 degree heating.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.And the negative control product do not show the speckle of same color on corresponding position.
3.3 aconitine limit is got this product 72ml (being equivalent to Radix Aconiti Kusnezoffii Preparata 3g), puts in the tool plug triangular flask water bath method, add ammonia solution 10ml, placed 1 hour, 150ml adds diethyl ether, close plug shakes up, and lets slip night, filter the medicinal residues 80ml that adds diethyl ether, supersound extraction 15min, filter, medicinal residues reuse ether is washed 3 times, each 15ml, filter, washing liquid and filtrate merge, the low temperature evaporate to dryness.Residue adds 1%HCl liquid 20ml, filters, and filtrate adds ammonia solution and regulates pH10~11, and ether (20ml * 2) jolting washing merges ether solution, and evaporate to dryness, residue add dehydrated alcohol 1ml, and dissolving is as need testing solution.Other gets the aconitine reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.Do not contain the negative sample of Radix Aconiti Kusnezoffii Preparata medical material again by the recipe quantity preparation, the preparation method of shining need testing solution prepares negative controls.According to thin layer chromatography (Chinese Pharmacopoeia version appendix in 2000 VIB) test, draw each 10 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, subnatant with chloroform-ethyl acetate-ammonia (10: 10: 2) is developing solvent, launches, and takes out, dry, spray is with the improvement bismuth potassium iodide test solution.In the test sample chromatograph, with reference substance chromatograph relevant position on, the speckle of appearance should or speckle not occur less than the speckle of reference substance.
One, antiinflammatory action experiment
Green pepper rhizome of chuanxiong spray gives the mice auricular concha local continuous 2 externals to be higher than clinical 1 times of liquor strength, normal clinical liquor strength, to be lower than clinical 1 times of liquor strength, and it is swollen significantly to suppress mice auricular concha caused by dimethylbenzene xylene; Give the rat left and right sides metapedes sole of the foot local continuous 2 externals, can significantly suppress rat Ovum Gallus domesticus album pedal swelling.
1 experiment purpose
Green pepper rhizome of chuanxiong spray is the effective preparation of a clinical treatment arthralgia pain, for verifying its curative effect and inquiring into its effect characteristics, has observed the antiinflammatory action of green pepper rhizome of chuanxiong spray.
2 experiment materials
2.1 experimental drug green pepper rhizome of chuanxiong spray is provided by auxilliary core medical sci-tech development company; Dimethylbenzene, reagent four Kunshan subsidiary factories of factory in Shanghai produce; The Shaolin bonesetting is smart, and Foshan Dezhong Pharmaceutical Co., Ltd. produces.
2.2 the laboratory animal mice, the Kunming kind, rat, SD provides by Henan Province's medical experiment animal center.
2.3 experimental apparatus PV-200 rat paw analyzer, Tai Sheng Science and Technology Ltd. in Chengdu produces
3 experimental techniques and result
3.1 to the bullate influence of mice auricular concha caused by dimethylbenzene xylene
Get 50 of body weight 18~21g male mices, evenly be divided into 5 groups at random, be respectively big, in, low dose of green pepper rhizome of chuanxiong spray group, smart group of Shaolin bonesetting and vehicle group, be coated with greatly to mouse right ear respectively, in, (high dose group is that drug level is higher than 1 times of clinical application [2g/ml] to low dose of green pepper rhizome of chuanxiong spray, middle dosage group is normal clinical medication [1g/ml], small dose group is in 1 times of clinical application at the bottom of the drug level, the dosage medicinal liquid is with composing with dilution agent 1 times [0.5g/ml] promptly), boneset smart original liquid and of Shaolin with the excipient (ethanol 30%) of volume, consumption is 0.1ml, all tow sides medications.Behind the medication 10min, clean auris dextra, immediately to mouse right ear drip be coated with 0.04ml dimethylbenzene (tow sides all are coated with) cause swollen, 10min after giving dimethylbenzene, every group of the same method of mouse right ear is coated with relative medicine once more.4h after giving dimethylbenzene, mice takes off cervical vertebra puts to death, and cuts ears, and alignment is laid left and right sides auricle same area with the 8mm card punch, weighs on analytical balance rapidly, and calculates the swelling degree.Swelling degree=auris dextra weight-left ear is heavy.The results are shown in Table 2 (one).
Table 2 (one) green pepper rhizome of chuanxiong spray is to the bullate influence of mice auricular concha caused by dimethylbenzene xylene
* represent with the excipient matched group than P<0.01
Can find out that from last table with excipient matched group ratio, the green pepper rhizome of chuanxiong spray group of big or middle dosage and the smart group of Shaolin bonesetting all can significantly reduce mice auricular concha caused by dimethylbenzene xylene swollen (P<0.01), low dose of green pepper rhizome of chuanxiong spray group has the bullate trend of the mice auricular concha caused by dimethylbenzene xylene of inhibition.Be that the external of green pepper rhizome of chuanxiong spray can significantly suppress the swelling of mice auricular concha.
3.2 influence to rat Ovum Gallus domesticus album pedal swelling
Get 50 of body weight 180~200g male rats, be divided into 5 groups at random, in sufficient sole of the foot analyzer, survey the normal left and right sides of each Mus metapedes sole of the foot volume before the experiment earlier, and be adjusted in the scribing position of rat ankle joint, make rat normal foot sole of the foot volume about 1ml.Then the rat left and right sides metapedes sole of the foot is immersed in greatly, in, (heavy dose of group drug level is higher than 1 times of clinical application [2g/ml] in the low dose of green pepper rhizome of chuanxiong spray medicinal liquid, middle dosage group is normal clinical medication [1g/ml], the small dose group drug level is lower than 1 times of clinical application, the dosage group is used (excipient dilutes 1 times [0.5g/ml]) promptly, Shaolin boneset (ethanol 30%) in smart original liquid and the vehicle group, consumption is the 1ml/ foot, it is 10min that the foot sole of the foot soaks the medicine time, allow the free movable 20min of rat, then in left and right sides metapedes plantar subcutaneous injection 10% fresh-laid egg clear liquid 0.1ml/ foot, 10min behind the albumen injection, again the rat left and right sides metapedes sole of the foot is immersed in 10min in the corresponding medicinal liquid, after giving Ovum Gallus domesticus album 0.5,1,2,4,6,8h, in sufficient sole of the foot analyzer, survey respectively and respectively organize rat left and right sides metapedes sole of the foot volume, and calculate toes swelling degree.The results are shown in Table 2 (two).
(2) can find out from table 2, and each organizes no explicitly difference between the rat normal foot sole of the foot volume, illustrate that grouping evenly.With excipient matched group ratio, dosage green pepper rhizome of chuanxiong spray group big or middle 30~480min after medication all can significantly suppress also sole of the foot swelling of rat Ovum Gallus domesticus album (P<0.01), low dose of green pepper rhizome of chuanxiong spray group 30min, 120~360min after medication can obviously suppress rat Ovum Gallus domesticus album pedal swelling (P<0.05), can significantly suppress the clear property of rat pedal swelling (P<0.01) at 60min; The smart group of Shaolin bonesetting 30~360min after medication can significantly suppress rat Ovum Gallus domesticus album pedal swelling (P<0.01), can obviously suppress rat Ovum Gallus domesticus album pedal swelling (P<0.05) at 480min.
4 brief summaries
The external of green pepper rhizome of chuanxiong spray is swollen to mice auricular concha caused by dimethylbenzene xylene, rat Ovum Gallus domesticus album pedal swelling all has better inhibitory action, and promptly green pepper rhizome of chuanxiong spray has better antiinflammatory action.
Two, analgesic activity experiment
Green pepper rhizome of chuanxiong spray can significantly alleviate the pain that formaldehyde causes the mice foot sole of the foot to be higher than clinical 1 times of liquor strength, normal clinical liquor strength, to be lower than clinical 1 times of liquor strength to 1 external of the mice foot sole of the foot, makes the pain prolongation of latency, and pain symptom alleviates; Can significantly improve hot plate method and cause the threshold of pain of mice pain.
1 experiment purpose
Green pepper rhizome of chuanxiong spray is the effective hospital of a clinical treatment arthralgia pain system, for verifying its effect and inquiring into its effect characteristics, has observed the analgesic activity of green pepper rhizome of chuanxiong spray.
2 experiment materials
2.1 experimental drug green pepper rhizome of chuanxiong spray is provided by auxilliary core medical sci-tech development company; Formaldehyde, solvent factory in Shanghai produces; The Shaolin bonesetting is smart, and Foshan Dezhong Pharmaceutical Co., Ltd. produces.
2.2 the laboratory animal mice, the Kunming kind is provided by Henan Province's medical experiment animal center.
2.3 experimental apparatus YLS-6A type mice hot plate analyzer, medical science equipment station, Shandong produces.
3 experimental techniques and result
3.1 the mice formaldehyde method is caused the influence of pain
Get 60 of body weight 18~21g mices, male and female half and half are divided into 5 groups at random.Respectively the right back foot of each group mice is immersed in greatly, in, (the high dose group drug level is higher than 1 times of clinical application [2g/ml] to low dose of green pepper rhizome of chuanxiong spray, middle dosage group is normal clinical medication [1g/ml], the small dose group drug level is lower than 1 times of clinical application, dosage dilutes 1 times [0.5g/ml] with excipient promptly), Shaolin boneset (ethanol 30%) in smart original liquid and the excipient, dosage is the 0.2ml/ foot, soaking the medicine time is 10min, allow the free movable 5min of mice recover normal then, give the formalin 30 μ l of the right back instep of mice subcutaneous injection 2.5% new preparation immediately, every mice of immediate record licks first to sting to lick in right back sufficient incubation period and the 5min to sting to lick in right back sufficient number of times and the 10min and stings right back sufficient number of times, the results are shown in Table 3.
Table 3 green pepper rhizome of chuanxiong spray causes the influence of pain to the mice formaldehyde method
* represent with the excipient matched group than P<0.05** represent with the excipient matched group than P<0.01
Can find out from last table, with excipient matched group ratio, heavy dose of green pepper rhizome of chuanxiong spray group can obviously prolong the incubation period (P<0.05) that formaldehyde method causes mice foot sole of the foot pain, can significantly reduce the interior mice of 5min and 10min and lick the number of times (P<0.01) of stinging the sufficient sole of the foot, in, the smart group of low dose of green pepper rhizome of chuanxiong spray group and Shaolin bonesetting can reduce significantly all that mice licks the number of times (P<0.01) of stinging the right back sufficient sole of the foot in 5min and the 10min, but PARA FORMALDEHYDE PRILLS(91,95) causes prolongation trend is only arranged the incubation period of mice foot sole of the foot pain.Be that green pepper rhizome of chuanxiong spray has good analgesic activity.
3.2 the mice hot plate method is caused the influence of pain
Get body weight 18~21g female mice, (55 ± 0.5 ℃) screen the normal threshold of pain of mice on hot-plate instrument earlier, all threshold of pains are lower than 3sec (pain sensation is too responsive) or are higher than all discarding of 30sec (pain sensation is insensitive) need not, select 60 of the qualified mices in the threshold of pain, by the threshold of pain mice is divided into 5 groups at random, every group 12, respectively the two metapedes of each group mice are immersed in greatly, in, (heavy dose of group drug level is higher than 1 times of clinical application [2g/ml] to low dose of green pepper rhizome of chuanxiong spray, middle dosage group is normal clinical medication [1g/ml], the small dose group drug level is lower than 1 times of clinical application, dosage dilutes 1 times [0.5g/ml] with excipient promptly), the Shaolin bonesets by (ethanol 30%) in smart original liquid and the excipient matched group, and dosage is the 0.2ml/ foot.Soaking the medicine time is 10min, allows the movable 5min of mice recover normal then, and 10min, 30min, 60min, 90min, 120min after medication measure the mice threshold of pain once more at hot-plate instrument respectively.Calculate threshold of pain improvement value, the different time threshold of pain after the improvement value=administration of the threshold of pain-with the preceding threshold of pain of Mus administration, all threshold of pains improvement value is that the negative person is by 0.The results are shown in Table 4.
Table 4 green pepper rhizome of chuanxiong spray causes the influence of mice pain to hot plate method
* represent with the excipient matched group than P<0.05, * * represent with the excipient matched group than P<0.01.In the bracket is threshold of pain improvement value.
Can find out that from last table each is organized between the normal threshold of pain of mice than no significant difference, illustrate that grouping evenly.With excipient matched group ratio, heavy dose of green pepper rhizome of chuanxiong spray group 10min, 30min after medication can significantly improve the mice threshold of pain (P<0.01), can obviously improve the mice threshold of pain (P<0.05) at medication 60min, 90min, 120min; Middle dosage green pepper rhizome of chuanxiong spray group 10~90min after medication can significantly improve the mice threshold of pain (P<0.01); Low dose of green pepper rhizome of chuanxiong spray group 10~30min after medication can obviously improve the mice threshold of pain (P<0.05); The smart group of Shaolin bonesetting 10~30min after medication can significantly improve the mice threshold of pain (P<0.01), can obviously improve the mice threshold of pain (P<0.05) at medication 60min, 90min, 120min.Act as excellently with the heavy dose group, the external application analgesic effect is held time in 90min.
4 brief summaries
Green pepper rhizome of chuanxiong spray external PARA FORMALDEHYDE PRILLS(91,95) method causes mice pain, hot plate method causes the mice pain all better analgesic activity.
Three, to the influence of rat assist agent arthritis
Green pepper rhizome of chuanxiong spray gives the rat left and right sides metapedes sole of the foot local continuous 21 externals being higher than clinical 1 times of liquor strength, normal clinical liquor strength, being lower than clinical 1 times of liquor strength, can significantly suppress pedal swelling due to the rat assist agent arthritis.
1 experiment purpose
Green pepper rhizome of chuanxiong spray is the effective hospital preparation of a clinical treatment arthralgia pain, for verifying its curative effect and inquiring into its effect characteristics, has observed the influence to adjuvant arthritis of green pepper rhizome of chuanxiong spray.
2 experiment materials
2.1 experimental drug green pepper rhizome of chuanxiong spray is provided lot number 050416 by Anyang iron and steel group company worker hospital general; Dimethylbenzene, reagent four Kunshan subsidiary factories of factory in Shanghai produce, lot number 020727; The Shaolin bonesetting is smart, and Foshan Dezhong Pharmaceutical Co., Ltd. produces, lot number 050601; Lanoline (chemical pure), Chinese Shanghai Huating lanoline factory, lot number 20020115; Liquid paraffin (chemical pure), Tianjin good fortune chemical reagent in morning factory produces, lot number 020401; Bacillus calmette-guerin vaccine, (BCG): 100mg/ props up, and lot number 0505001 is provided by Shanghai Vaccine and Serum Institute.
2.2 experimental animal rat, Wistar, male, Hebei province's Experimental Animal Center, cleaning level, the rat quality certification number, the moving word 410116 of Henan doctor.
2.3 experimental apparatus PV-200 rat paw analyzer, Tai Sheng Science and Technology Ltd. in Chengdu produces
3 experimental techniques and result
Get 40 of Mus great and mighty or powerful, be divided into 5 groups at random, all make the adjuvant arthritis model, method for give the sufficient lift in a rat left side down the Freud. Freund's complete adjuvant of the new preparation of injection (getting 4 parts of heat fused of anhydrous lanolin is placed in the mortar, cools off slightly, adds liquid paraffin to 6 part while grinding, autoclave sterilization, press 5mg/ml again and add bacillus calmette-guerin vaccine, grind well, be Freund's complete adjuvant).In sufficient sole of the foot analyzer, survey the normal left and right sides of each Mus metapedes sole of the foot volume before the experiment earlier, and be adjusted in the scribing position of rat ankle joint, make rat normal foot sole of the foot volume about 1ml.Promptly began administration the same day in injection adjuvant, (it is that normal clinical medication [1g/ml], small dose group drug level are lower than 1 times of clinical application that heavy dose of group drug level is higher than clinical application 1 times of [2g/ml], middle dosage group to use the green pepper rhizome of chuanxiong spray of large, medium and small dosage at left toes respectively, the dosage group bonesets by (ethanol 30%) in smart original liquid and the vehicle group with (excipient dilution 1 times [0.5g/ml]), Shaolin promptly, and consumption is 1ml/ enough.Absorbent cotton is adsorbed corresponding 1ml medicinal liquid, absorbent cotton embraced gently apply on the toes of the rat left and right sides, use immobilization with adhesive tape after external one deck cellophane wrapped, make medicinal liquid contact 1 hour with the rat toes, remove then and be subjected to the reagent thing, every toes medicinal liquid 1ml, administration every day 1 time, continuous use 21 days cutd open rat extremely on the 21st day.Respectively at reaching back 3 hours of injection before the injection adjuvant, 2nd, in sufficient sole of the foot analyzer, surveyed in 4,6,9,12,15,18,21 days and respectively organize the left back sufficient sole of the foot volume of rat, surveyed in sufficient sole of the foot analyzer in the 9th, 12,15,18,21 day and respectively to organize the right back sufficient sole of the foot volume of rat, calculate toes swelling degree.The results are shown in Table 5, table 6.
Can find out from table 5, corresponding time ratio with the excipient matched group, the smart group of dosage green pepper rhizome of chuanxiong spray group big or middle and Shaolin bonesetting all can be after giving adjuvant significantly suppresses the swelling of rat paw due to the adjuvant in the 2nd day, 4 days, 6 days, 8 days, 12 days, 15 days, 18 days, 21 days, make the pedal swelling rate significantly reduce (P<0.01), heavy dose of green pepper rhizome of chuanxiong spray group makes the pedal swelling rate obviously reduce (P<0.05) in the swelling that administration 3h can obviously suppress rat paw due to the adjuvant; Low dose of green pepper rhizome of chuanxiong spray group obviously suppressed the swelling of rat paw due to the adjuvant in the 2nd day, 4 days, 6 days, 8 days, 12 days, 15 days, 18 days, 21 days after giving adjuvant, make the pedal swelling rate obviously reduce (P<0.05).
Can find out from table 6, corresponding time ratio with the excipient matched group, big or middle dosage green pepper rhizome of chuanxiong spray and the smart group of Shaolin bonesetting are being used adjuvant the 12nd~21 day, can obviously suppress the Secondary cases swelling (P<0.05) of the rat right side foot sole of the foot.
4 brief summaries
The external of green pepper rhizome of chuanxiong spray has better inhibitory action to pedal swelling due to the rat assist agent arthritis, can obviously suppress partial early stage inflammatory reaction of injection adjuvant and the swelling once again after 12 days, obviously suppress the Secondary cases foot swelling of offside toes again, promptly green pepper rhizome of chuanxiong spray has better antiinflammatory action.
Four, clinical experimental data:
Clinical data: patient's 40 examples, male 22 examples, women 18 examples, the oldest 65 years old, minimum 32 years old; Cervical vertebra ache 6 examples wherein, lumbar vertebra pain 5 examples, the gonalgia 29 examples.Typical clinical manifestation is all arranged: joint " rest pain " is stiff, and be movable dumb.Cervical vertebra ache causes symptoms such as numbness of the fingers and dizziness; Symptoms such as bony crepitus can appear in the gonalgia.The course of disease is the longest 20 years, and is the shortest 3 years.
Therapeutic Method: spray is sprayed on sore spot, spray in per 1 hour 1 time, the spray consumption is there to be obvious liquid in the part; 7d-10d was 1 course of treatment.But as 1 course of treatment of DeGrain reuse.Should withdraw all analgesic drug products during the treatment and when changing dressings, some little red pimples occur, illustrate to reach drug effect as the discovery disease sites.
Criterion of therapeutical effect: effectively: pain disappears, and function of joint is improved; Produce effects: pain relief, function of joint obtain part to be improved; Invalid: the same with symptom before the treatment.
Therapeutic outcome: the course of treatment the shortest 7d, the longest 20d.Effective 27 examples account for 67.5%: produce effects 11 examples account for 27.5%; Invalid 2 examples account for 5.00%.Total effective rate is 95.00%.
In a word, clearly show that spray of the present invention has damp eliminating cold expelling, relaxing muscles and tendons and activating QI and blood in the collateral, blood circulation promoting and blood stasis dispelling, reducing swelling and alleviating pain by above-mentioned data, bone joint pain etc. had the good curing effect, good effect, effective percentage height, having no side effect, is the innovation on the Chinese medicine external curing bone joint pain medicine.