CN106822377B - Compound osteitis cataplasm patch and preparation method thereof - Google Patents
Compound osteitis cataplasm patch and preparation method thereof Download PDFInfo
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- CN106822377B CN106822377B CN201710204257.2A CN201710204257A CN106822377B CN 106822377 B CN106822377 B CN 106822377B CN 201710204257 A CN201710204257 A CN 201710204257A CN 106822377 B CN106822377 B CN 106822377B
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- 238000004018 waxing Methods 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
- 239000009750 xian ling gu bao Substances 0.000 description 1
Classifications
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/11—Pteridophyta or Filicophyta (ferns)
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- A61K36/18—Magnoliophyta (angiosperms)
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- A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
- A61K9/7023—Transdermal patches and similar drug-containing composite devices, e.g. cataplasms
- A61K9/703—Transdermal patches and similar drug-containing composite devices, e.g. cataplasms characterised by shape or structure; Details concerning release liner or backing; Refillable patches; User-activated patches
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- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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Abstract
The invention provides a compound osteitis cataplasm patch and a preparation method thereof. The cataplasm is prepared from twelve medicinal materials of rhizoma drynariae, valeriana jatamansi, eucommia ulmoides, rhizoma cibotii, asiatic toddalia root, caulis spatholobi, caulis periplocae, herba epimedii, caulis perllae, and caulis sargentodoxae on the basis of fully excavating a Miao medicine fumigation proved recipe.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a compound osteitis cataplasm and a preparation method thereof.
Background
Osteoarthritis is one of the common diseases in clinic, and can cause osteonecrosis in later period and dysfunction of hip joint, thus seriously affecting the life quality and labor capacity of patients. The femoral head necrosis patients in China are 500-800 ten thousand, and more than 15 ten thousand new cases per year. The disease has high clinical treatment cost, long period and unsatisfactory curative effect, and brings great burden to patients and society.
Miao ethnic group medicine fumigation therapy is a representative external treatment method of Miao ethnic group medicine, Miao ethnic group medicine fumigation therapy is a method for treating diseases by fumigating diseased parts of patients with water vapor evaporated by medicine, and medicinal effective components can enter the body along with medicine vapor through respiratory tract and expanded pores. The Miao medicine is good in fresh medicine treatment, the Miao medicine fumigation therapy combines the medicine therapy and the thermal therapy, the capillary vessels of the skin and the muscles of the affected part can be expanded by the thermal therapy, the blood circulation is accelerated, the excretion of vivotoxin can be accelerated by matching with the medicine, the metabolism is promoted, the obstacles such as the skin, the ligaments, the tendons, the bones and the blood vessels are relieved, and the normal function of the human body is recovered, so that the treatment purpose is achieved.
However, the traditional Miao fumigation therapy also has the following problems in clinical use: the operation of the apparatus is inconvenient, the skin of a patient is directly contacted with the fumigation instrument, cross infection is easy to occur, and the selection of time and place is inconvenient, so that the treatment of the patient is difficult, and the popularization and the application are difficult. In addition, the Miao medicine fumigation proved recipe is mainly a folk experience prescription, the dosage of the medicine formula is mainly based on experience, scientific and reasonable optimization is lacked, the clinical application mainly aims at auxiliary treatment, and the Miao medicine value cannot be better exerted.
Cataplasma is an external emplastrum, and has a long history of application in japan. Is prepared by mixing the extract of medicinal materials, medicinal materials or/and chemical drugs with appropriate hydrophilic matrix, and coating on backing material to obtain patch, which comprises backing (common non-woven fabric, elastic cloth), ointment, and anti-mucosa (isolating membrane on surface of ointment). However, due to factors such as high selection requirements on medicines (especially traditional Chinese medicine extract) and matrixes, difficulty in screening, high research cost and the like, the development of the cataplasm is directly restricted, and in recent years, with the rapid development of pharmaceutical academia and application thereof at home and abroad, the unique medication simplicity and safety of a transdermal drug delivery system make the transdermal drug delivery system become one of the most remarkable fields in the drug development of the 21 st century.
Therefore, the invention aims to overcome the defects of fumigation therapy, exert the curative effect to the maximum extent, develop folk proved recipes to provide an effective, safe and convenient treatment method and medicament for osteoarthritis patients, develop the cataplasm which not only retains the efficacy of the traditional Miao medicine fumigation therapy, but also is economic and convenient, fully exert the potential value of Miao medicine and have important significance for the development of Miao medicine.
Disclosure of Invention
In order to solve the technical problems, the invention provides a compound osteitis cataplasm patch and a preparation method thereof. The cataplasm is prepared on the basis of fully excavating a Miao medicine fumigation proved recipe, has a rigorous formula structure and a proper and simple preparation process, overcomes the defects of a Miao medicine fumigation method in clinical application, has large drug-loading rate, high drug utilization degree and quick response, and has a very remarkable effect of treating osteoarthritis.
Specifically, the technical scheme of the invention is as follows:
a compound cataplasma for treating osteitis is mainly prepared from the following substrates and traditional Chinese medicine extracts:
matrix: the material is prepared by blending 7-9 parts of polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (soluplus), 7-9 parts of gelatin, 5-7 parts of sodium polyacrylate, 7-9 parts of titanium dioxide, 5-7 parts of sodium carboxymethylcellulose, 20-40 parts of polyvinylpyrrolidone, 1-2 parts of tartaric acid, 0.1-1 part of dihydroxyaluminum glycinate, 180 parts of glycerol 140 and 50-90 parts of distilled water.
Traditional Chinese medicine extract: is prepared by extracting 5-10 parts of rhizoma drynariae, 5-10 parts of valeriana jatamansi jones, 5-10 parts of eucommia bark, 5-10 parts of cibotium barometz, 5-10 parts of asiatic toddalia root, 5-10 parts of suberect spatholobus stem, 5-10 parts of periploca forrestii, 5-10 parts of epimedium herb, 5-10 parts of lysimachia christinae hance, 5-10 parts of lycopodium clavatum and 5-10 parts of sargentgloryvine stem.
Further, the compound osteitis cataplasm is mainly prepared from the following matrixes and traditional Chinese medicine extracts:
matrix: the adhesive is prepared by blending 8 parts of polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (soluplus), 8 parts of gelatin, 6 parts of sodium polyacrylate, 8 parts of titanium dioxide, 6 parts of sodium carboxymethylcellulose, 30 parts of polyvinylpyrrolidone, 1.5 parts of tartaric acid, 0.5 part of dihydroxyaluminum glycinate, 160 parts of glycerol and 70 parts of distilled water in parts by weight;
traditional Chinese medicine extract: the traditional Chinese medicine composition is prepared by extracting 8 parts by weight of rhizoma drynariae, 8 parts by weight of valeriana jatamansi jones, 8 parts by weight of eucommia ulmoides, 8 parts by weight of cibotium barometz, 8 parts by weight of asiatic toddalia root, 8 parts by weight of suberect spatholobus stem, 8 parts by weight of periploca forrestii, 8 parts by weight of epimedium herb, 8 parts by weight of caulis perllae, 8 parts by weight of common clubmoss.
Further, the proportion of the matrix to the traditional Chinese medicine extract in the compound osteitis cataplasm is 10: 1-5; preferably, the ratio of the matrix to the Chinese medicinal extract is 10: 3.
The traditional Chinese medicine extract is prepared by the following steps:
weighing 12 medicinal materials of rhizoma drynariae, valeriana jatamansi jones, eucommia ulmoides, cibotium barometz, asiatic toddalia root, suberect spatholobus stem, periploca forrestii, epimedium, lysimachia christinae hance, lycopodium clavatum and sargentgloryvine stem, adding 8 times of water, decocting for 2 times, 1.5 hours each time, filtering, recovering filtrate, concentrating, drying, crushing and sieving with a 200-mesh sieve to obtain the traditional Chinese medicine extract.
The invention also provides a preparation method of the compound osteitis cataplasm patch, which comprises the following steps:
precisely weighing sodium polyacrylate and titanium dioxide, mixing, adding glycerol, and grinding to obtain phase A. Precisely weighing carboxymethylcellulose sodium, gelatin, polyvinylpyrrolidone, and polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (soluplus), mixing, adding water, and adding in oil bath to make it fully swell to obtain phase B. A, B grinding the two phases, mixing, adding tartaric acid and aluminum glycoxide, adding the Chinese medicinal extract, and mixing. Coating the cataplasm on a back lining, and airing to obtain the cataplasm.
The invention has the beneficial effects that:
firstly, in the pharmacodynamic research process of the traditional Miao medicine fumigation therapy and the classic Miao medicine fumigation proved recipe, the problems that most of the prescriptions are handed down from experience, the formula dosage is combined randomly, scientific verification is lacked, the clinical efficacy is not obvious enough, the efficacy is slow to take effect and the like are caused, and the Miao medicine fumigation therapy is mainly taken as auxiliary treatment in clinic at present. Therefore, the invention carries out the re-matching of primary and secondary medicaments, and finally determines the prescription as follows through a plurality of times of prescription screening: rhizoma Drynariae, radix Pimpinellae Candolleanae, Eucommiae cortex, rhizoma Cibotii, radix Toddaliae Asiaticae, caulis Spatholobi, caulis Seu radix Schisandrae Henryi, caulis et folium Periplocae Forrestii, herba Epimedii, caulis et folium Gaultheriae Yunnanensis, herba Lycopodii, and caulis Sargentodoxae. Rhizoma drynariae: nature and taste: bitter and warm. Enters kidney and liver meridians, and has the effects of tonifying kidney, strengthening bone, healing wound and relieving pain. Valeriana jatamansi jones: 2, pungent taste; slightly bitter; warm in nature, regulate qi-flowing for regulating the middle warmer, dispel cold and remove dampness; promoting blood circulation and relieving swelling. Eucommia ulmoides: it is sweet in taste and warm in nature. Has effects in tonifying liver and kidney, strengthening muscle and bone, regulating Chong and ren meridians, and preventing miscarriage. Rhizoma cibotii: bitter, sweet and warm. Tonify liver and kidney, strengthen waist and spine, dispel wind-damp. Asiatic toddalia root: dispelling pathogenic wind, relieving pain, removing blood stasis, and stopping bleeding; caulis spatholobi: the properties and tastes of the herbs are bitter, sweet and warm. It enters liver and kidney meridians. The main treatment is blood enriching, blood circulation promoting and vein relaxing. Caulis spatholobi: the main functions are promoting qi circulation, relieving pain, promoting blood circulation and removing blood stasis; black-bone vine: promote blood circulation to restore menstrual flow, dispel wind-damp. Herba Epimedii: herba Epimedii can tonify kidney yang, strengthen tendons and bones, and dispel wind-damp. Bone penetrating incense: dispel wind and dampness, activate blood and dredge collaterals. B, common clubmoss herb: dispel wind and cold, remove dampness and relieve swelling, relax tendons and activate collaterals. Sargentgloryvine stem: detoxicating, resolving carbuncle, promoting blood circulation, dredging collaterals, dispelling pathogenic wind, and killing parasite; the formula adopts a formula concept of three monarch drugs, three minister drugs, three adjuvant drugs and three guide drugs, rhizoma drynariae, eucommia bark and herba epimedii are used as monarch drugs, caulis periplocae, lycopodium clavatum and caulis spatholobi are used as minister drugs, a co-assistant drug is Wei, rhizoma cibotii, jatamans valeriana rhizome and caulis spatholobi are used as adjuvant drugs, and asiatic toddalia root, Tokyo and sargentgloryvine stem are used as conductant drugs to lead the drugs to float upwards, the twelve drugs supplement each other, coordinate and enhance the effects and achieve the drug effects together, and the cataplasm is large in drug property and very suitable for being prepared. According to the invention, a large number of screening tests are carried out on the adaptability of the cataplasm matrix and the traditional Chinese medicine extract, and the finally prepared cataplasm is large in drug-loading rate, high in bioavailability, very obvious in relation to treatment of osteoarthritis, and convenient and simple in test.
To further illustrate the advantageous effects of the present invention, the present invention also provides the following test examples, which are intended to illustrate the advantageous effects of the present invention, but in no way limit the contents of the present invention:
firstly, screening a prescription:
the prescription of the invention is added or subtracted on the basis of a classical 'Miao medicine fumigation proved prescription', the invention respectively uses twelve medicines of the prescription of the invention, a ministerial medicine component, an adjuvant medicine component and a messenger medicine component to prepare a cataplasm for pharmacodynamic investigation, and inspects 2012 and 2016 years, patients in orthopedics clinics and wards of a second subsidiary hospital of the Guiyang traditional Chinese medical college are diagnosed as early femoral head necrosis and accord with the inclusion standard, and the patients are randomly divided into four groups of a normal treatment group and a cataplasm group according to the admission sequence. Normal treatment group treatment regimen: 6ml/d of compound bone peptide and 60ml/d of breviscapine are infused into the solution by vein, and the Xianlinggubao is orally taken for 3 tablets/time and 3 times/d, after 3 weeks of treatment, the solution is rested for 1 week and is continuously treated for 3 months. Treatment regimen of babu group: the cataplasm treatment is added on the basis of the normal treatment scheme, 1 time/d, and the treatment time is the same as that of the normal treatment group. Before and after treatment, the clinical evaluation table and the grade judgment standard of the patient are respectively applied to carry out comprehensive evaluation on the treatment, and the clinical curative effect of the cataplasm is summarized and compared.
The results show that: in contrast to the normal treatment group: the excellent rate of the twelve-medicine composition is improved by 43 percent, the excellent rate of the ministerial medicine group is improved by 15 percent, the excellent rate of the adjuvant medicine group is 20 percent, and the excellent rate of the messenger medicine group is 22 percent.
Secondly, screening by a babu plaster process:
in the preparation process of the traditional Chinese medicine extract cataplasm, a better cataplasm matrix formula is screened by adopting the prior art for preparation, and the quality evaluation is carried out:
test group 1 was prepared using a matrix formulation NP700 crosslinker [ al (oh)3 EDTA-2Na ALCL3] glycerol 0.75(0.02:0.01:0.02): 10.
Test group 2: the preparation is prepared from a matrix formula of carbomer, namely sodium polyacrylate, namely polyvinylpyrrolidone K30, and sodium carboxymethyl cellulose, namely polyvinyl alcohol, namely 3.0:4.0:0.5:0.2: 0.5.
Test group 3: the matrix formula is adopted: 0.8g of gelatin, 0.6g of sodium polyacrylate, 0.8g of titanium dioxide, 16g of glycerol, 0.6g of sodium carboxymethylcellulose, 3g of polyvinylpyrrolidone, 0.15g of tartaric acid and 0.05g of dihydroxyaluminum glycolate.
Test group 4: the matrix formula is adopted: 0.8g of polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (soluplus), 0.8g of gelatin, 0.6g of sodium polyacrylate, 0.8g of titanium dioxide, 16g of glycerol, 0.6g of sodium carboxymethylcellulose, 3g of polyvinylpyrrolidone, 0.15g of tartaric acid and 0.05g of dihydroxyaluminum glycolate.
Investigation and evaluation method: the cataplasm was evaluated by using initial adhesion, holding adhesion, and sensory evaluation (appearance, skin comfort, residual ointment, skin followability, repeated uncovering) as evaluation indexes. Finally, all indexes are integrated to obtain a comprehensive score, and the quality of the compound osteomyelitis cataplasm prepared from different matrixes is evaluated according to the score, which is shown in table 1.
TABLE 1 quality evaluation of Compound GUYAN cataplasma
| Evaluation index | Method of producing a composite material | Index score |
| Initial adhesion | First method of adhesion determination method of XIIE emplastrum in appendix of Chinese pharmacopoeia | 0～25 |
| Holding power of adhesion | Method for measuring adhesive force of XIIE emplastrum in appendix of Chinese pharmacopoeia | 0～25 |
| Appearance character | Uniform appearance, no insoluble particles and bubbles | 0～10 |
| Skin comfort | Moderate hardness, no tension and no irritation | 0～10 |
| Residual property of paste | After the plaster is removed, the plaster surface is kept intact, and no residue is left on the skin | 0～10 |
| Skin following property | After being stuck on the wrist, the wrist is thrown forcefully for 10 times without falling off | 0～10 |
| Repeated stripping property | After repeatedly peeling off, good viscosity can be maintained | 0～10 |
The results are shown in Table 2 below:
table 2 investigation of test results
| Test group | Initial adhesion | Holding power of adhesion | Sensory evaluation | Composite score |
| Test group 1 | 18 | 17 | 35 | 70 |
| Test group 2 | 20 | 20 | 36 | 76 |
| Test group 3 | 22 | 22 | 40 | 84 |
| Test group 4 | 24 | 23 | 47 | 94 |
The test results show that the invention screens out a better cataplasm substrate formula in the prior art to prepare cataplasms (test group 1 and test group 2), the quality evaluation comprehensive score of the cataplasms is lower, the quality of the cataplasms is poorer, the preparation of the substrate (test group 3) preferably selected by the invention is obviously improved compared with that of the test group 1-2, but the effect is still not ideal enough, and the in vitro dissolution test result is only 72%; in order to meet the requirements of the cataplasm well, the invention carries out analysis and research on the extract, and unexpectedly finds that after the extract is dried, crushed and sieved by a 200-mesh sieve, and the polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (soluplus) is added into a cataplasm substrate, the dissolution rate of the cataplasm is greatly improved, the in-vitro dissolution rate test result is 92%, and the prepared cataplasm is comprehensively evaluated for 94 minutes and has optimal quality.
Thirdly, pharmacodynamics and related researches of the compound osteitis cataplasm:
(1) anti-inflammatory experimental study of compound osteitis babu plaster
1 materials of the experiment
1.1 medicaments
Test drugs osteitis remedy prepared in examples 1-5 of the present invention, 3cm × 3 cm; positive control drug: the four-side tablet muscle and bone pain patch is produced by four-side pharmaceutical industry group Limited company in Henan province, the batch number is 1260019, and the specification is 7cm multiplied by 10 cm; the functions and indications are as follows: relieving pain caused by femoral head necrosis, hyperostosis, cervical syndrome, lumbar vertebra disease, intervertebral disc prolapse, rheumarthritis, ankylosing spondylitis, scapulohumeral periarthritis, traumatic injury and injury, senile osteoarthropathy, etc., and promoting its rehabilitation.
1.2 Experimental animals
Kunming mouse, male and female half, weight 18-22g, license number SCXK 20120006. 30 male rats with common wistar, the weight of 200-: chongqing city experimental animal detection center.
1.3 Experimental reagents and instruments
Sodium sulfide: henjin chemical reagents manufacturing Co., Ltd, lot number 20120221 (8% solution prepared with 75% ethanol just before use); xylene Chongqing metallocene chemical reagent, Inc. batch No. 20130508; a puncher (6mm), a timer and an electronic balance; micrometer calipers.
2 method of experiment
2.1 Effect of Compound osteitis cataplasm on swelling degree of mouse auricle caused by xylene
40 mice were randomly divided into 5 groups of 8 mice (half female and half male), and 24 hours prior to the experiment were abdomen dehaired with 8% sodium sulfide, vehicle control: 0.3ml of 30% ethanol is taken per dose; positive control group: the plaster is 3cm × 3cm for each time; compound osteitis cataplasm high dose group: each patch was given the cataplasm patch prepared in example 5 of the present invention; the bone inflammation diminishing cataplasm medium dosage is as follows: the cataplasma prepared in the 2-4 steps of the invention embodiment is given; cataplasm low dose group: the cataplasm prepared in example 1 of the present invention was administered. The preparation is administered 1 time per day for 3 days, and each group is administered 6 hours per day via abdominal administration. 1 hour after the last administration, the right ear of the mouse was coated with 0.05 ml/mouse of xylene, and the left ear was used as a control. After 30 minutes, the cervical vertebrae were removed to kill the mice, the left and right auricle lugs were cut off, circular lugs at the same positions were taken from both auricles with a 6 mm-diameter punch, and the obtained weights were weighed, and the difference between the drug group and the vehicle control group was compared with each other by using the difference in weight between the two lugs as an index of swelling degree, and the inhibition ratio was determined.
Swelling degree (mg) ═ weight of right ear tablet (mg) — weight of left ear tablet (mg)
The swelling inhibition ratio (%) was (average swelling degree in the excipient group-average swelling degree in the administered group)/average swelling degree in the excipient group x 100%.
2.2 Effect of Compound osteitis cataplasm on degree of swelling of foot sole
Healthy wistar male rats were taken 30 and randomly divided into 5 groups of 6, vehicle control groups: 0.3ml of 30% ethanol is taken per dose; positive control group: the plaster is 3cm × 3cm for each time; compound osteitis cataplasm high dose group: each patch was given the cataplasm patch prepared in example 5 of the present invention; the bone inflammation diminishing cataplasm medium dosage is as follows: the cataplasma prepared in the 2-4 steps of the invention embodiment is given; cataplasm low dose group: the cataplasm prepared in example 1 of the present invention was administered. The preparation is administered 1 time per day for 3 days, and each group is administered 6 hours per day via abdominal administration. The thickness of the right hind paw of each rat was measured on day 3 as the normal value before molding, and after 1 hour of the last dose, 0.1ml of 10% fresh egg white was subcutaneously injected to the hind paw of each animal in each group to create an acute paw edema model, and the thickness of the same part of the hind paw was measured with a micrometer caliper at 0.5, 1, 2,4 and 6 hours after inflammation, respectively. The difference of the diameters of the footpads before and after inflammation of each rat is the swelling degree, and the swelling degree is taken as a detection index. The difference between the swelling degree of the drug-administration group and the swelling degree of the excipient control group was compared, and statistical analysis was performed.
Swelling degree (mm) is the diameter of the metatarsal of the foot after inflammation-the diameter of the metatarsal of the foot before inflammation.
3 data processing and statistical method
The experimental data are expressed by Mean +/-standard deviation (Mean +/-SD), and SPSS (Statistical product and Service Solutions, SPSS)19.0 Statistical analysis software is adopted for mathematical Statistical analysis, in the experiment of influence of the compound osteitis cataplasm on swelling degree of mouse auricles caused by dimethylbenzene, each group of experimental data are subjected to normality test by a Shapiro-Wilk method and a Kolmogrov-Smitnov method, then the homogeneity of variance is analyzed by a Leven method, if the experimental data accord with the normal distribution and the homogeneity of variance, variance analysis is adopted for comparison of multiple groups of data, LSD-t test is adopted for comparison of multiple groups, the significance of variance analysis is alpha 0.05, and P <0.05 shows that the Statistical significance is achieved. In the experiment of the influence of the compound osteitis babu plaster on the degree of swelling of the foot sole, the variance analysis of multiple groups of repeated measurement design data is adopted for each group of experimental data, and P <0.05 shows that the statistical significance is achieved.
4.1 Effect of Compound osteitis cataplasm on swelling degree of mouse auricle caused by xylene
In an anti-inflammatory experiment, the medium-dose group and the positive control group of the cataplasm have certain inhibition effects on mouse auricle swelling caused by xylene, and the differences of the medium-dose group and the positive control group have statistical significance compared with an excipient control group. The anti-inflammatory effect of the medium-dose group of the cataplasm patch is greater than that of the positive control group, and the medium-dose of the cataplasm patch can obviously resist inflammatory reaction caused by dimethylbenzene. See table 3.
TABLE 3 Effect of Compound Guyan babu plaster on swelling of mouse auricle caused by xylene: (
n＝8)
Note: each group was compared to the vehicle group, P < 0.05.
4.2 Effect of Compound osteitis cataplasm on degree of swelling of foot sole
Under the test level of alpha being 0.05, the result of the spherical test is not satisfied (P is less than 0.01), so that the data information (table 2) is subjected to variance analysis by using a Spss19.0 repeated measurement special module. The Huynh-Feldt test result shows that: the interaction between the treatment effect and the time effect has no statistical significance (F is 1.891, P is 0.52), the time main effect has a statistical significance (F is 67.832, P is less than 0.0001), the treatment main effect also has a statistical significance (F is 5.009, P is less than 0.01), and the results of pairwise comparison of the effects among the groups show that: the difference between the low dose group of the cataplasm patch and the excipient control group is not statistically significant (P >0.05), the difference between the middle dose group of the cataplasm patch and the excipient control group is statistically significant (P <0.05), and the difference between the middle dose group of the cataplasm patch and the positive control group of the cataplasm patch and the excipient control group is statistically significant (P <0.01), which indicates that the middle dose group has obvious inhibition effect on the vola swelling of rats caused by egg white and is superior to the high and low dose groups. See Table 4
TABLE 4 Effect of Fufang Guyan babu Patch on egg white swollen degree (mm) of rat: (
n＝8)
Swelling degree of foot: f group is 5.009, and P is less than 0.01; f time is 67.832, and P is less than 0.01; and F interaction is 1.891, and P > 0.05. (2) Analgesic pharmacodynamics research of compound osteitis cataplasm
1 materials of the experiment
1.1 medicaments
1 materials of the experiment
1.1 medicaments
Test drugs osteitis remedy prepared in examples 1-5 of the present invention, 3cm × 3 cm; positive control drug: the four-side tablet muscle and bone pain patch is produced by four-side pharmaceutical industry group Limited company in Henan province, the batch number is 1260019, and the specification is 7cm multiplied by 10 cm; the functions and indications are as follows: relieving pain caused by femoral head necrosis, hyperostosis, cervical syndrome, lumbar vertebra disease, intervertebral disc prolapse, rheumarthritis, ankylosing spondylitis, scapulohumeral periarthritis, traumatic injury and injury, senile osteoarthropathy, etc., and promoting its rehabilitation.
1.2 Experimental animals
40 Kunming mice, each half of male and female, the weight is 18-22g, and the issuing unit: chongqing city experimental animal detection center. License number SCXK (Yu) 20120006.
1.3 Experimental reagents
Sodium sulfide: henjin chemical reagents manufacturing Co., Ltd, lot number 20120221 (8% solution prepared with 75% ethanol just before use); glacial acetic acid: national drug group chemical laboratory limited, lot number T20110106. CJ-8402 type hot plate pain measuring instrument, Zhejiang Ninghaibaishi electronic medical instrument factory.
2. Experimental methods
2.1 Effect of cataplasma Patches on writhing response in mice
The grouping and application method and time of 40 mice are the same as the anti-inflammatory experiment. After the topical drug was washed with warm water, each mouse was injected with 0.1ml 10g-1 of 0.6% glacial acetic acid (acetic acid solution was prepared just before use) to induce writhing reaction, and the writhing frequency of each mouse was recorded within 15 minutes after injection of pain, and each administration group was compared with the vehicle group.
2.2 Effect of Compound osteitis cataplasm on Hot plate pain in mice
A plurality of female mice are taken, the mice are placed on a hot plate instrument with the temperature of (55 +/-0.5) DEG C one by one, the pain threshold value of the mice is measured and recorded, 40 mice with the pain threshold value within 5-30 s are selected by taking licked feet as the pain index, the mice are randomly divided into 5 groups, and each group comprises 8 mice. Then, the mice were placed one by one on a hot plate apparatus, the pain threshold was measured 2 times each (at intervals of 10min), and the average value was taken as the basic pain threshold of the mice, i.e., the normal pain threshold before administration. The mice of each group were belly-dehaired with 8% sodium sulfide and each day belly dosed separately, vehicle control group dosed with 30% ethanol 0.3 ml/mouse; the positive control group is a square plaster of 3cm × 3cm for muscle and bone pain each time;
the pain threshold of the mice was determined 30min after the last administration and was taken as the post-administration pain threshold. If the mice were still not algesic within 60s on the hotplate, the pain threshold increase before and after administration was calculated and counted as 60 s.
Percent increase in pain threshold (%) (pain threshold after administration-basal pain threshold)/basal pain threshold × 100%
3 data processing and statistical method
The experimental data are expressed by Mean +/-standard deviation (Mean +/-SD), and are subjected to mathematical Statistical analysis by SPSS (Statistical product and Service Solutions, SPSS)19.0 Statistical analysis software, each group of experimental data are subjected to normality test by a Shapiro-Wilk method and a Kolmogrov-Smitnov method, and then are subjected to homogeneity of variance analysis by a Leven method, if the normal distribution and the homogeneity of variance are met, the multiple groups of data are compared by variance analysis, the multiple groups of data are compared by an LSD-t test, the significance of the difference analysis is 0.05, and P <0.05 shows that the Statistical significance is achieved.
4 results
4.1 Effect of Compound Guyan cataplasm on mouse writhing response
In an analgesic experiment, the low, medium and high dose groups of the compound osteitis cataplasm patch are different from the excipient group, and have statistical significance (p is less than 0.05). The results show that each group can resist the pain reaction caused by glacial acetic acid stimulation of mice, so that the times of mouse body twisting are obviously reduced, and the dosage group has the optimal effect. See table 5.
TABLE 5 Effect of Compound Guyan cataplasm on analgesic action of mice (wriggle method) (wriggle method)
n＝8)
| Group of | n | Number of wriggling within 15min |
| Control group of excipients | 8 | 40.88±3.36 |
| Positive control group | 8 | 23.75±2.49* |
| Cataplasm high dose group | 8 | 28.25±2.12* |
| Cataplasm medium dose group | 8 | 19.38±2.67* |
| Cataplasm low dose group | 8 | 29.50±2.07* |
Note: each group was compared to the vehicle group, p < 0.05.
4.2 Effect of Compound osteitis cataplasm patch on hot plate-induced pain of mice
In an analgesic experiment, both the high and medium dose groups after administration increased the pain threshold in mice compared to the vehicle group, with the difference having statistical significance (P < 0.05). And the analgesic effect of the medium-dose group of the cataplasm is strongest. Compared with a group with high and medium dose of the cataplasma before self administration, the pain threshold of the mice can be improved to different degrees, all have significant difference and have statistical significance (P < 0.05). See table 6.
TABLE 6 Effect of FUFANGGUYANBABU Patch on analgesic effect of mice (Hot plate method) (for hot plate)
n＝8)
Note: each group was compared to the vehicle group: p < 0.05; comparison with pre-dose pain threshold: delta P < 0.05.
(3) Establishment of rabbit osteoarthritis model and influence of compound osteitis cataplasm on tissue morphology
1 materials of the experiment
1.1 Experimental animals
70 healthy adult New Zealand white rabbits with the weight of 2.0 plus or minus 0.5kg in each half of the male rabbit and the female rabbit are selected. Issuing unit: chongqing city experimental animal detection center. License number SCXK (Yu) 20120006.
1.2 control of the Experimental conditions
The temperature of a laboratory room is kept at about 26 ℃, the humidity is 50% -60%, ventilation is kept, and during the experiment, only one single rabbit is used for each group, and the light is normally collected.
1.3 feed
The rabbit feed is fed by a special rabbit feed provided by Guiyang Chinese medicine college animals, and water is freely drunk.
1.4 drugs and Main laboratory Equipment
Sodium sulfide: tianjin Hengxing Chemicals manufacturing Co., Ltd, Lot 20120221 (prepared as an 8% solution with 75% ethanol just before use).
② compound osteitis cataplasm: cataplasms prepared according to examples 1-5 of the present invention. The specification of each patch is 3cm multiplied by 3cm provided by the preparation laboratory of Guiyang traditional Chinese medicine college.
And thirdly, the compound osteomyelitis cataplasm blank matrix is prepared by blending and coating water, sodium carboxymethylcellulose, NP-700, glycerol, azone, talcum powder, gelatin and the like on non-woven fabrics according to a proportion, placing the non-woven fabrics in a blast constant temperature dryer, slightly drying the non-woven fabrics and covering the non-woven fabrics with a preservative film, wherein the prepared compound osteomyelitis cataplasm matrix is semitransparent milky white. The specification of each patch is 3cm multiplied by 3 cm.
Fourthly, positive contrast medicine: the four-side tablet muscle and bone pain patch is produced by the four-side pharmaceutical industry group ltd of Henan province, the batch number is 1260019, and the specification is 7cm multiplied by 10 cm: the functions and indications are as follows: relieving pain caused by femoral head necrosis, hyperostosis, cervical syndrome, lumbar vertebra disease, intervertebral disc prolapse, rheumarthritis, ankylosing spondylitis, scapulohumeral periarthritis, traumatic injury, senile osteoarthropathy, etc., and promoting its rehabilitation.
The rabbit hutch box is provided by the laboratory of animal institute of traditional Chinese medicine of Guiyang.
HH-W digital display constant temperature water bath.
Seventy-seven (B) 05002584y ultrasonic atomizer for fish jumping
And eighthly, an embedding machine.
Ninthly CELLSENS STANDAR D biological microscopic image analyzer
And cutter for LEICA RM 2235.
2 method of experiment
2.1 grouping of Experimental animals
70 healthy New Zealand white rabbits. The male and female halves were each 2.0. + -. 0.5kg in body weight and were randomly divided into 7 groups. Each group had 8. The method comprises (10 model groups), a normal control group, a positive control group, a square muscle and bone pain patch, a Miao medicinal fumigation group, a compound osteitis cataplasm high dose group, a compound osteitis cataplasm medium dose group, and a compound osteitis cataplasm medium dose group.
2.2 making of Experimental animal models
Except for the normal group, the other groups adopt the plaster cast fixation of the left hind limb ankle joint extension position from the first week to the sixth week of the experiment, the length of the plaster cast is that the upper end is close to the middle-upper 1/3 of the tibia, and the lower end is close to the 1/3 of the foot, and the fixation is continued for 6 weeks. After 6 weeks, the model group was sacrificed 2, articular cartilage was taken, gross specimens and pathological changes of cartilage were observed, left and right were separately marked, and 10% neutral formalin solution was placed for fixation and examined. So as to know the joint braking effect. After the molding is finished, the plaster is removed for fixation, and the left hind limb ankle cartilage is taken as a specimen for pathological observation after the rest (except the normal group) is treated for 6 weeks.
2.3 groups of administration methods
Normal group: feeding normally without administration of medicine.
Model group: the compound osteitis cataplasm blank matrix is given.
Positive control group: the plaster is administered with 3cm × 3cm for each time.
Miao medicine fumigation therapy group: the vaccine concentrate is fumigated once a day, half an hour each time.
The fumigation method comprises the following steps:
firstly, the concentrated fumigant solution is put into a container containing the fumigant of the B05002584y fish jumping ultrasonic atomizer, and a spraying pipe is put into a water bath kettle.
Controlling the temperature of the water in the HH-W digital display constant temperature water bath kettle to be 60 ℃.
Thirdly, the spraying pipe is connected with the rabbit hutch box after passing through the water bath pot, and the temperature of the chemical vapor is controlled to be 35-40 ℃.
And fourthly, putting the experimental animal rabbit into a rabbit cage box, and fumigating for half an hour. The rabbit box is covered with cotton cloth to prevent the medicine vapor from diffusing, so as to avoid affecting the treatment effect.
Compound osteitis cataplasm high dose group: the cataplasm prepared in example 5 of the present invention was administered.
The compound osteomyelitis cataplasm middle dose group comprises: the cataplasma prepared in examples 2 to 4 of the present invention was administered.
Compound osteitis cataplasm low dose group: the cataplasm prepared in example 1 of the present invention was administered.
The above groups were administered once a day for 6 hours continuously for 6 weeks.
2.4 Observation index
2.4.1 general Observation
Observing the mental state, activity condition, feeding, diarrhea, hair loss, body surface infection, survival condition and the like of the experimental animal, weighing the weight of the experimental animal once a week to observe the change of the weight of the experimental animal.
2.4.2 Observation of gross specimens
All animals were fasted for 12h after 6 weekend treatment, and the experimental animals were sacrificed by ear-margin venous air embolization. The hair is cut off and disinfected at the ankle joint to be tested, the skin, the superficial fascia and the deep fascia are sequentially incised by a scalpel, the ankle joint cavity is exposed, and the general changes of the shape, the color, the thickness and the like of the joint cavity and the surface of the articular cartilage are observed by naked eyes. Articular cartilage was scored according to literature standard for gross observation: 0 minute: the joint surface is flat and the color is normal; 1 minute: the joint surface is rough, has cracks and is dark in color; and 2, dividing: joint surface erosion, cartilage defect reaches cartilage surface and middle layer; and 3, dividing: joint surface ulcer is formed, and cartilage defect reaches the deep layer of cartilage; and 4, dividing: the cartilage is stripped and the subchondral bone is exposed.
2.4.3 Observation of cartilage Pathology
Using a scalpel to draw a fixed point at the cartilage of the ankle joint surface of the experimental side to reach the cartilage lower layer deeply, trimming the cartilage to 0.5cmx0.3cmx 0.2cm, cleaning the cartilage with 0.9% of physiological saline, placing the cartilage in 10% of neutral formic acid solution for fixing for one day, and decalcifying the fixed cartilage for one month at normal temperature (changing the fluid once in 3 days) by 10% of EDTA (PH-7). Specimen is dehydrated regularly (70% alcohol for half an hour → 80% alcohol for 1 hour 20 minutes → 90% alcohol for 1 hour → 95% alcohol for 45 minutes → 100% alcohol for 30 minutes), so as to form a gradient dehydration; conventional transparence (putting bone tissues into 100% alcohol respectively: the xylene is 1: 1 solution for 20 minutes → xylene (first 30 minutes → xylene) (second 30 minutes); conventional waxing (respectively soaking bone tissues into a wax jar for 1 hour → a wax jar for 1 hour); routine paraffin embedding (4 um thick serial sections, serial number, then Hematoxylin-eosin (HE) staining, finally complete cartilage full-layer). The pathological changes of the articular cartilage are scored according to the Mankin' S scoring standard, and the tissue structure, the cell number and the tide line integrity of the articular cartilage are observed in each section. The higher the score obtained, the more severe the lesion. See table 7.
TABLE 7 Mankin' S Scoring standards
2.4.4 bone histomorphometry Studies
According to the cartilage pathological sections, the thickness of the cartilage is observed by using an CELLSENS STANDAR D biological microscopic image analyzer, 4 visual fields are randomly selected from each section, and the thickness of the ankle joint cartilage of each group is measured and photographed.
2.4.5 bone histomorphometry Studies
4 high power visual fields (200 times) are randomly selected from each cartilage section, wherein 2 positions are located on a cartilage surface layer and a cartilage transitional layer, and the other 2 positions are located on a cartilage radiation layer and a cartilage calcified layer, and attention is paid to the fact that 4 layer areas which are adjacent to a degeneration defect part but have complete cartilage structures are selected for observation. The total number of chondrocytes in the 4-layer region was counted.
3 data processing and statistical method
All the experimental data are expressed by Mean +/-standard deviation (Mean +/-SD), and are subjected to mathematical statistical analysis by SPSS (statistical product and Service Solutions, SPSS)19.0 statistical analysis software, each group of experimental data are subjected to normality test by a Shapiro-Wilk method and a Kolmogrov-Smitnov method, and then are subjected to homogeneity of variance analysis by a Leven method, if the normal distribution and the homogeneity of variance are met, the multiple groups of data are compared by variance analysis, the multiple groups of data are compared by an LSD-t test, the significance of the difference analysis is alpha 0.05, and P <0.05 shows that the statistical significance is achieved.
4 results
4.1 general observations:
the animals in the normal group had good mental status, sensitive response, free movement of the ankle joint and glossy hair color during the experiment. All the groups after the molding have limp to different degrees, the motion of ankle joints is obviously limited, the spirit is low, and the luster is poor. After the compound osteitis cataplasm high and medium dose groups are treated, the degree is relieved, and the medium dose effect is most obvious.
4.2 gross macroscopic observations of rabbit ankle joint specimens:
normal control group: the articular cartilage has smooth surface, regular edges, milk white color, high transparency, no cracks and defects, clear and bright synovial fluid and less quantity.
Model control group: the articular cartilage has rough and uneven surface, dark color, grayish yellow color and obvious small cracks; the molding of varying size, morphology and depth can result in thinning and fragmentation of cartilage. Hyperemia and edema of the synovium; joint fluid is increased and the color is pale yellow.
Miao medicine fumigation group: the joint surface is light white, the surface is smooth, and cracks are formed on the surface of part of the specimen.
Compound osteitis cataplasm low dose group: the surface of the articular cartilage loses luster and is slightly grayish yellow, obvious cracks exist, and the wear degree of the cartilage is slightly reduced compared with that of a model control group.
The compound osteomyelitis cataplasm middle dose group and the positive control group are as follows: the articular cartilage is yellow-white, the surface part loses normal luster, the color is slightly dark and smooth, and obvious erosive cartilage defect is not seen. The difference between the two groups was not apparent by visual inspection. The appearance and color of the two groups of articular cartilage are better than those of the model group. The results are shown in Table 8.
TABLE 8 score table of total articular cartilage at 6 weeks (n. 4) for each group
| Group of | Number of animals | 0 point (min) | 1 minute (1) | 2 is divided into | 3 points of | 4 is divided into |
| Normal group | 4 | 4 | 0 | 0 | 0 | 0 |
| Model set | 4 | 0 | 0 | 1 | 1 | 2 |
| Positive control group | 4 | 0 | 1 | 2 | 1 | 0 |
| Miao medicine fumigation group | 4 | 0 | 1 | 1 | 2 | 0 |
| Cataplasm high dose group | 4 | 0 | 1 | 2 | 1 | 0 |
| Cataplasm medium dose group | 4 | 0 | 1 | 2 | 2 | 0 |
| Cataplasm low dose group | 4 | 0 | 0 | 1 | 2 | 1 |
4.3 results of articular cartilage observation under optical microscope:
normal group: the articular cartilage structure is normal, the cells are arranged regularly, the cell nucleus is dark blue, and the cytoplasm is pink. The surface layer cartilage cells are fusiform and similar to fiber cells; the cells in the middle layer are circular and irregularly arranged; the columnar cell layer is arranged in a columnar shape, and tide lines can be seen. The cartilage matrix is uniformly colored and pink.
Model control group: the thickness of cartilage becomes thin, the number of cells is reduced, the arrangement is disordered, the number of cracks is more, part of cartilage reaches a calcified layer, particularly, the number of superficial cells is mainly reduced, the cartilage is covered by more inflammatory cells, the density of cells in the cartilage is uneven, and the tide line is disordered.
Positive control group: cartilage structures exist, cells are arranged regularly, and the tide lines are clear. The cartilage matrix is uniformly colored and pink.
The medium dose group: the coloring of the cartilage matrix is uniform, the layering of the cells is clear and ordered, the cell morphology is regular, and the tide lines are clear.
High and low dose groups: a few fissures appear on the cartilage surface, partly reaching the irradiated layer. The cells are not well-leveled, irregularly arranged and incompletely affected by the fuzzy tide lines. But better than the model set.
Miao medicine fumigation group: the coloring of the cartilage matrix is uniform, the cells are arranged in a column shape, the layers are clear, the cell morphology is regular, and the tide lines are clear. The results are shown in Table 9.
TABLE 9 eachGroup cartilage Mankin' S score results (
n＝4)
| Group of | n | Average integral (point) |
| Model set | 4 | 10.00±0.82★ |
| Positive drug group | 4 | 6.00±0.82▲ |
| Miao medicine fumigation group | 4 | 7.25±0.96▲★ |
| Compound osteitis cataplasm high-dose group | 4 | 6.25±0.96▲ |
| Compound osteitis cataplasm medium-dose group | 4 | 5.25±0.96▲ |
| Compound osteitis cataplasm low-dose group | 4 | 8.75±0.96▲★ |
Note: comparing the solidup-p of each group with the model group is less than 0.05; comparison of ≤ p in positive drug group is 0.05.
4.4 cartilage thickness statistics
Each group had a significant difference (p <0.05) compared to the model group. The medium dose group has no statistical difference (p <0.05) with the positive control group, which indicates that the medium dose group of the compound osteomyelitis cataplasm has better treatment effect with the positive control group in preventing and treating the thickness of the articular cartilage of osteoarthritis. The results are shown in Table 10.
TABLE 10 statistics of ankle cartilage thickness table (
n＝4)
Note: comparing each group with the model group, wherein tangle-solidup-p is less than 0.05; comparison with the positive drug group, p < 0.05.
4.5 chondrocyte count statistics
Compared with the model group, the low-dose group of the compound osteitis cataplasm has no significant difference (p > 0.05). Each of the other groups had a significant difference (p <0.05) compared to the model group. The ankle cartilage cells of the medium dose group and the positive control group were compared. The compound osteitis cataplasm shows that the medium-dose group of the compound osteitis cataplasm has better treatment effect on preventing and treating the osteoarthritis and increasing the number of chondrocytes compared with a positive control group. The results are shown in Table 11.
Table 11 statistics of chondrocyte counts table (
n＝4)
Note: comparing the groups with the model group, wherein a-solidup-p is less than 0.05; comparison with the positive drug group, p < 0.05.
(4) Discussion of the related Art
The experiment of inflammation caused by auricle edema of a mouse is that the right ear is smeared with an inflammation-causing medicine, namely dimethylbenzene, and the left ear is used as a control to cause the local capillary blood vessel congestion, permeability increase, exudation increase and edema, which show the symptoms of red, swelling, heat, pain and the like of the auricle of the mouse. The experiment on inflammation caused by swelling degree of foot sole of a rat is that egg white is injected into the foot sole of the rat to cause local acute inflammation and local tissue edema. The inhibition of the drug on the swelling degree of the hind foot was observed. Is a molding method for acute inflammation. The hot plate test of mice is a special reaction of foot addition when the feet of the mice are stimulated by hot plate to cause pain. The pain threshold value is the time required after the hot plate is contacted with the hot plate and the feet are added, and the pain threshold value change between the medicine group and the control group is compared by taking the time as an observation index to verify the analgesic effect of the medicine. The writhing experiment of the mouse is a sensitive, simple and good-repeatability method for screening the effect of the non-steroidal anti-inflammatory drug, and the model is suitable for the research of central and peripheral analgesics. In the experiment, acetic acid is injected into the abdominal cavity of the mouse to cause lasting pain reaction of the abdominal cavity, so that the mouse has writhing reaction. Many studies have shown that the effect strength of the drug and the clinical analgesic effect have an obvious correlation when the model is used for studying the analgesic effect of the drug. The 4 inflammation and analgesia models selected in the research are classic test methods for verifying the anti-inflammatory and analgesic effects of the medicine, and can be used for researching the anti-inflammatory and analgesic effects of the compound osteitis cataplasm.
The test result shows that: the effect of the invention is obvious, the positive control group has excellent effect, the effect of treating osteoarthritis is very obvious and is superior to the high and low dose groups, which shows that the preferable traditional Chinese medicine formula and the substrate formula of the compound osteitis cataplasm are scientific and reasonable.
Toxicology of compound osteitis cataplasm and related research
1 materials and methods
1.1 materials
Selecting 72 healthy Wistar rats and 30 white guinea pigs (experimental animals are provided by Chongqing Tengxin biotechnology, Inc.), self-preparing compound osteitis cataplasm and a compound osteitis cataplasm matrix,
1.2 methods
30 healthy Wistar rats with half male and female bodies and weight of 200-250g are selected for acute toxicity test and randomly divided into two groups, namely 20 in the group A and 10 in the group B. Rats were dehaired on their backs with 8% sodium sulfide 24h prior to the experiment. The group A is pasted with a large dose of compound osteitis cataplasm patch, and the group B is pasted with a compound osteitis cataplasm matrix for 24 h. After 24h, the drug was washed off and the rats were observed for 2 weeks according to the index of table one.
TABLE 12 animal reactivity index observed in acute toxicity test
The skin irritation test selects 42 Wisar natural rats with half weight and 200-250g weight, and the rats are randomly divided into 7 groups, namely a complete skin high-dose group, a complete skin medium-dose group, a complete skin low-dose group, a damaged skin high-dose group, a damaged skin medium-dose group, a damaged skin low-dose group and a negative control group, wherein each group comprises 6 rats. Rats were dehaired on their backs with 8% sodium sulfide 24h prior to the experiment. Rats were dehaired on their backs with 8% sodium sulfide 24h prior to the experiment. The administration is continued for 14 days, and local skin irritation is observed for 30-60min, 24h, 48h, and 72h after the last administration. And scoring according to the standard of the table two, and evaluating according to the table three.
TABLE 13 skin irritation response Scoring criteria
TABLE 14 evaluation criteria for skin irritation Strength
The skin active allergy test selects 30 healthy white guinea pigs with half male and female, 6-8 weeks old and weight of 250-300g, and randomly divides the guinea pigs into 3 groups of 10 guinea pigs. Guinea pigs were deprived of flank hair 24h prior to the trial with 8% sodium sulfide. The test group and the negative control group were 0.2 g/mouse, the positive group was 0.2 ml/mouse, and the drugs were administered 1 time each on days 0, 7, and 14 of the sensitization phase and 28 of the challenge phase. The test group was given compound osteitis cataplasm, the negative control group was given compound osteitis cataplasm matrix, and the positive control group was given 2, 4-dinitrochlorobenzene, with a sensitization period concentration of 1% and an excitation period concentration of 0.1% (prepared with 70% ethanol). The drug is administered to the left depilatory part in the sensitization phase and to the right depilatory part in the stimulation phase. After excitation, observation was carried out for 6, 24, 48, 72 h. The scores were scored according to the criteria of Table four and the evaluations were made in Table five.
TABLE 15 Scoring criteria for skin allergy
TABLE 16 evaluation criteria for skin allergy
2 results
During the acute toxicity test, general behavioral signs of animals in each group are uniform, local skin at the administration part is normal, and no local and systemic toxicity reaction of rats is observed; the big tolerance of the rats to the osteomyelitis is more than 21.5 g/kg.
During the skin irritation test period, general behavioral signs and local skin at the administration part of each group of animals are not abnormal, and obvious irritation reactions such as skin edema, erythema and the like are not seen in the broken skin and the intact skin groups; according to the skin irritation evaluation standard, the skin irritation evaluation result is shown in the sixth table, and the compound osteitis cataplasm has no irritation to the skin of a rat. The compound osteitis cataplasm has no obvious irritation to the damaged and intact skin of rats. However, long-term continuous administration of the drug may have some effect on wound healing.
TABLE 17 skin irritation results of Compound GUYAN cataplasma
During the active skin allergy test period of the guinea pig, the general behavioral signs of the animals in the test group and the negative control group and the local skin at the administration position are not abnormal, the general behavioral signs of the animals in the positive control group are normal, and erythema and incrustation are occasionally generated on the local skin in different degrees. The scoring result is shown in that after the compound osteitis babu plaster and the skin of the guinea pig are pasted for multiple times, no obvious allergic reaction such as skin erythema, edema and the like appears after one guinea pig is found, on the contrary, all the guinea pigs of the positive control group have skin erythema, edema and even scab reaction with different degrees after being excited, the skin reaction integral is 5.9, the sensitization rate is 100 percent, and the skin reaction is extreme sensitization; the compound osteitis cataplasm has no obvious sensitization on guinea pig skin.
TABLE 18 evaluation results of compound osteitis cataplasma guinea pig active skin allergy test
Discussion of 3
The research results of three toxicological tests, namely an acute toxicity test, a skin irritation test and a skin active allergy test, show that the compound osteitis cataplasm belongs to a skin external application drug, is only used on the skin, has low toxicity after transdermal administration, has no toxic reaction on the intact skin or the damaged skin, has no influence on the whole body physiological function of animals, and is a high-safety external application drug for treating the osteitis.
The specific implementation mode is as follows:
example 1: preparation of babu plaster for treating osteitis
(1) Weighing 4g of rhizoma drynariae, 4g of valeriana jatamansi, 4g of eucommia ulmoides, 4g of cibotium barometz, 4g of toddalia asiatica, 4g of caulis spatholobi, 4g of periplocae forrestii, 4g of caulis perllae margaritae, 4g of lycopodium clavatum and 4g of sargentgloryvine stem, adding 8 times of water, decocting for 2 times, 1.5 hours each time, filtering, recovering filtrate, concentrating, drying, crushing and sieving with a 200-mesh sieve to obtain the traditional Chinese medicine extract.
(2) Precisely weighing 4g of sodium polyacrylate and 6g of titanium dioxide, uniformly mixing, adding 130g of glycerol, and uniformly grinding to obtain a phase A. Precisely weighing 4g of sodium carboxymethylcellulose, 6g of gelatin, 18g of polyvinylpyrrolidone and 6g of polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (soluplus), uniformly mixing, adding 45g of water, and adding in an oil bath to fully swell to obtain a phase B. A, B, grinding and mixing evenly, then adding 0.1 times of the matrix of the traditional Chinese medicine extract obtained in the step (1), 0.8g of tartaric acid and 0.1g of dihydroxyaluminium, and mixing evenly. Coating the cataplasm on a back lining, and drying to obtain the cataplasm.
Example 2: preparation of babu plaster for treating osteitis
(1) Weighing 5g of rhizoma drynariae, 5g of valeriana jatamansi, 5g of eucommia ulmoides, 5g of cibotium barometz, 5g of toddalia asiatica, 5g of caulis spatholobi, 5g of periploca forrestii, 5g of herba epimedii, 5g of caulis perllae, 5g of lycopodium clavatum and 5g of sargentgloryvine stem, adding 7 times of water, decocting for 2 times, 1.5 hours each time, filtering, recovering filtrate, concentrating, drying, crushing and sieving with a 200-mesh sieve to obtain the traditional Chinese medicine extract.
(2) Precisely weighing 5g of sodium polyacrylate and 7g of titanium dioxide, uniformly mixing, adding 140g of glycerol, and uniformly grinding to obtain a phase A. Then precisely weighing 5g of sodium carboxymethylcellulose, 7g of gelatin, 20g of polyvinylpyrrolidone and 6g of polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (soluplus), uniformly mixing, adding 50g of water, and adding in an oil bath to fully swell to obtain a phase B. A, B, grinding and mixing evenly, adding 0.1 times of the matrix of the traditional Chinese medicine extract obtained in the step (1), 1g of tartaric acid and 0.1g of dihydroxyaluminium glycolate, and mixing evenly. Coating the cataplasm on a back lining, and drying to obtain the cataplasm.
Example 3: preparation of babu plaster for treating osteitis
(1) Weighing 8g of rhizoma drynariae, 8g of valeriana jatamansi, 8g of eucommia ulmoides, 8g of cibotium barometz, 8g of toddalia asiatica, 8g of caulis spatholobi, 8g of periplocae, 8g of herba epimedii, 8g of caulis perllae, 8g of lycopodium clavatum and 8g of sargentgloryvine stem, adding 8 times of water, decocting for 2 times, 1.5 hours each time, filtering, recovering filtrate, concentrating, drying, crushing and sieving with a 200-mesh sieve to obtain the traditional Chinese medicine extract.
(2) Precisely weighing 6g of sodium polyacrylate and 8g of titanium dioxide, uniformly mixing, adding 160g of glycerol, and uniformly grinding to obtain a phase A. Precisely weighing 6g of sodium carboxymethylcellulose, 8g of gelatin, 30g of polyvinylpyrrolidone and 8g of polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (soluplus), uniformly mixing, adding 70g of water, and adding in an oil bath to fully swell to obtain a phase B. A, B, grinding and mixing evenly, adding 0.3 times of the matrix of the traditional Chinese medicine extract obtained in the step (1), 1.5g of tartaric acid and 0.5g of dihydroxyaluminium, and mixing evenly. Coating the cataplasm on a back lining, and drying to obtain the cataplasm.
Example 4: preparation of babu plaster for treating osteitis
(1) Weighing 10g of rhizoma drynariae, 10g of valeriana jatamansi, 10g of eucommia ulmoides, 10g of cibotium barometz, 10g of toddalia asiatica, 10g of caulis spatholobi, 10g of periploca forrestii, 10g of herba epimedii, 10g of caulis perllae, 10g of lycopodium clavatum and 10g of sargentgloryvine stem, adding 9 times of water, decocting for 2 times, 1.5 hours each time, filtering, recovering filtrate, concentrating, drying, crushing and sieving with a 200-mesh sieve to obtain the traditional Chinese medicine extract.
(2) Precisely weighing 7g of sodium polyacrylate and 9g of titanium dioxide, uniformly mixing, adding 180g of glycerol, and uniformly grinding to obtain a phase A. Precisely weighing 7g of sodium carboxymethylcellulose, 9g of gelatin, 40g of polyvinylpyrrolidone and 9g of polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (soluplus), uniformly mixing, adding 90g of water, and adding in an oil bath to fully swell to obtain a phase B. A, B, grinding and mixing evenly, adding 0.5 times of the matrix of the traditional Chinese medicine extract obtained in the step (1), 2g of tartaric acid and 1g of dihydroxyaluminium glycolate, and mixing evenly. Coating the cataplasm on a back lining, and drying to obtain the cataplasm.
Example 5: preparation of babu plaster for treating osteitis
(1) Weighing 11g of rhizoma drynariae, 11g of valeriana jatamansi, 11g of eucommia ulmoides, 11g of cibotium barometz, 11g of toddalia asiatica, 11g of caulis spatholobi, 11g of periplocae, 11g of herba speranskiae tuberculatae, 11g of lycopodium clavatum and 11g of sargentgloryvine stem, adding 9 times of water, decocting for 1.5 hours each time, filtering, recovering filtrate, concentrating, drying, crushing and sieving with a 200-mesh sieve to obtain the traditional Chinese medicine extract.
(2) Precisely weighing 9g of sodium polyacrylate and 11g of titanium dioxide, uniformly mixing, adding 190g of glycerol, and uniformly grinding to obtain a phase A. Precisely weighing 8g of sodium carboxymethylcellulose, 10g of gelatin, 45g of polyvinylpyrrolidone and 10g of polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (soluplus), uniformly mixing, adding 100g of water, and adding in an oil bath to fully swell to obtain a phase B. A, B, grinding and mixing evenly, adding 0.6 times of the matrix of the traditional Chinese medicine extract obtained in the step (1), 2.2g of tartaric acid and 1.1g of dihydroxyaluminium, and mixing evenly. Coating the cataplasm on a back lining, and drying to obtain the cataplasm.
Claims (3)
1. A compound osteitis cataplasm is characterized in that: is prepared from the following substrates and traditional Chinese medicine extracts: matrix: according to parts by weight, the material is prepared from 7-9 parts of polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer, 7-9 parts of gelatin, 5-7 parts of sodium polyacrylate, 7-9 parts of titanium dioxide, 5-7 parts of sodium carboxymethylcellulose, 20-40 parts of polyvinylpyrrolidone, 1-2 parts of tartaric acid, 0.1-1 part of dihydroxyaluminum glycinate, 180 parts of glycerol 140 and 50-90 parts of distilled water; traditional Chinese medicine extract: the traditional Chinese medicine composition is prepared from, by weight, 5-10 parts of drynaria rhizome, 5-10 parts of valeriana jatamansi jones, 5-10 parts of eucommia ulmoides, 5-10 parts of cibotium barometz, 5-10 parts of toddalia asiatica, 5-10 parts of caulis spatholobi, 5-10 parts of sargentgloryvine stem, 5-10 parts of herba epimedii, 5-10 parts of caulis perllae, 5-10 parts of lycopodium clavatum and 5-10 parts of sargentgloryvine stem, and is prepared from a matrix and a traditional Chinese medicine extract in a mass ratio of 10:1-5, wherein the traditional Chinese medicine extract is prepared by the following steps: weighing rhizoma Drynariae, Valeriana jatamansi Jones, Eucommiae cortex, rhizoma Cibotii, radix Toddaliae Asiaticae, caulis Spatholobi, caulis Seu folium Schisandrae Henryi, caulis et folium Periplocae Forrestii, herba Epimedii, caulis et folium Gaultheriae Yunnanensis, herba Lycopodii, and caulis Sargentodoxae, adding 7-9 times of water, decocting for 2 times (1.5 hr each time), filtering, recovering filtrate, concentrating, drying, pulverizing, and sieving with 200 mesh sieve to obtain traditional Chinese medicine extract; the preparation method of the compound osteitis cataplasm patch comprises the following steps: precisely weighing sodium polyacrylate and titanium dioxide, uniformly mixing, adding glycerol, and uniformly grinding to obtain phase A; precisely weighing carboxymethylcellulose sodium, gelatin, polyvinylpyrrolidone, polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer, mixing uniformly, adding water, and adding in oil bath to make it fully swell to obtain phase B; grinding A, B phases, mixing, adding tartaric acid and aluminum glyceroxide, adding Chinese medicinal extract, and mixing; coating the cataplasm on a back lining, and airing to obtain the cataplasm.
2. The compound osteitis cataplasm of claim 1, which is characterized in that: is prepared from the following substrates and traditional Chinese medicine extracts: matrix: the adhesive is prepared from 8 parts by weight of polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer, 8 parts by weight of gelatin, 6 parts by weight of sodium polyacrylate, 8 parts by weight of titanium dioxide, 6 parts by weight of sodium carboxymethylcellulose, 30 parts by weight of polyvinylpyrrolidone, 1.5 parts by weight of tartaric acid, 0.5 part by weight of dihydroxyaluminium glycinate, 160 parts by weight of glycerol and 70 parts by weight of distilled water; traditional Chinese medicine extract: the traditional Chinese medicine composition is prepared by extracting 8 parts by weight of rhizoma drynariae, 8 parts by weight of valeriana jatamansi jones, 8 parts by weight of eucommia ulmoides, 8 parts by weight of cibotium barometz, 8 parts by weight of asiatic toddalia root, 8 parts by weight of suberect spatholobus stem, 8 parts by weight of periploca forrestii, 8 parts by weight of epimedium herb, 8 parts by weight of caulis perllae, 8 parts by weight of common clubmoss.
3. The compound osteitis babu plaster as claimed in claim 1 or 2, which is prepared from a matrix and a traditional Chinese medicine extract in a mass ratio of 10: 3.
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| 徐剑等.正交试验优化骨炎消巴布剂的基质处方.《中国实验方剂学杂志》.2012,第18卷(第24期),参见第39页左栏末段、右栏末段，第40页第2.2、2.4节. * |
| 正交试验优化骨炎消巴布剂的基质处方;徐剑等;《中国实验方剂学杂志》;20121231;第18卷(第24期);第39-41页，尤其是第39页左栏末段、右栏末段，第40页第2.2、2.4节 * |
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Address after: No. 50, Shidong Road, Nanming District, Guiyang City, Guizhou Province Patentee after: Guizhou University of Traditional Chinese Medicine Address before: No. 50, Nanming District, Nanming District, Guiyang, Guizhou Patentee before: GUIYANG College OF TRADITIONAL CHINESE MEDICINE |
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Application publication date: 20170613 Assignee: First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine Assignor: Guizhou University of Traditional Chinese Medicine Contract record no.: X2021520000001 Denomination of invention: A compound Guyan cataplasm and its preparation method Granted publication date: 20200925 License type: Common License Record date: 20210305 |