CN106075295B - Application of osteopathy and anesthesia medicine in preparing anti-inflammatory and analgesic medicine - Google Patents

Application of osteopathy and anesthesia medicine in preparing anti-inflammatory and analgesic medicine Download PDF

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CN106075295B
CN106075295B CN201610546385.0A CN201610546385A CN106075295B CN 106075295 B CN106075295 B CN 106075295B CN 201610546385 A CN201610546385 A CN 201610546385A CN 106075295 B CN106075295 B CN 106075295B
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吴广文
陈俊
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Fujian University of Traditional Chinese Medicine
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Abstract

The invention discloses application of the following raw material medicines in weight ratio in preparing anti-inflammatory and analgesic medicines, wherein the raw material medicines comprise the following components in weight ratio: 0.5 to 1.5 parts of ephedra, 0.5 to 1.5 parts of eggplant, 0.5 to 1.5 parts of turmeric, 0.5 to 1.5 parts of common monkshood mother root, 0.5 to 1.5 parts of kusnezoff monkshood root and 1 to 3 parts of Chinese azalea flower; and further provides a gel prepared by taking the compound as an active ingredient, and the gel can fully exert the anti-inflammatory and analgesic effects. The pharmacodynamic experiment shows that the raw material medicines in the weight ratio have obvious anti-inflammatory and analgesic effects, the effect is superior to that of a commercial medicine, namely nataline, the medicine has no obvious skin irritation, the safety is good, and the medicine has wide clinical application prospects.

Description

Application of osteopathy and anesthesia medicine in preparing anti-inflammatory and analgesic medicine
Technical Field
The invention relates to application of a bone-setting anesthetic in preparing an anti-inflammatory and analgesic drug, and belongs to the field of medicines.
Background
Inflammation is a defense response of the body to stimuli and is usually manifested by redness, swelling, heat, pain, dysfunction, and the like, including infectious and non-infectious inflammation. At present, the anti-inflammatory drugs commonly used in clinic are mainly divided into two main categories: one class is steroidal anti-inflammatory drugs, such as hydrocortisone and its artificially synthesized derivatives; another class is non-steroidal anti-inflammatory drugs, which are antipyretic, analgesic and anti-inflammatory drugs in medical practice such as aspirin, phenylbutazone, celecoxib, etc.
Pain is an unpleasant sensation and mood, a symptom of many diseases, that is associated with substantial and potential tissue damage, as well as physiological and psychological factors. In many cases, analgesic treatment of patients is required. There are two main classes of drugs commonly used for analgesia, analgesics that inhibit prostaglandin biosynthesis, and analgesics that interact with opioid receptors.
Long-term clinical practice shows that the anti-inflammatory and analgesic drugs commonly used at present have adverse reactions of different degrees, for example, aspirin has serious gastrointestinal side effects, and liver cell cholestasis and granulomatous hepatitis caused by phenylbutazone can cause death of some patients.
Therefore, the problem to be solved is to provide a substitute drug which has definite anti-inflammatory and analgesic effects and no obvious toxic or side effect.
The bone-setting anesthetic is from 'the secret medical prescription of record in Qingtai hospital, prescription and surgical injury gate', and the usage and dosage of the bone-setting anesthetic are recorded in the book: "you take the arrowhead five times when the tea and wine are taken, and there is no pain when the tea and wine are taken any time. Decoct gan Cao and pour it into decoction, and take it out. The formula medicine is taken orally for anesthesia in clinical traditional Chinese medicine.
At present, no report about the anti-inflammatory and analgesic effects of the osteopathy and anesthesia medicine is found.
Disclosure of Invention
The invention aims to provide the application of the osteopathy anesthetic in preparing anti-inflammatory and analgesic drugs.
The invention provides application of the following raw material medicines in preparing anti-inflammatory and analgesic medicines, wherein the raw material medicines comprise the following components in parts by weight: 0.5 to 1.5 parts of ephedra, 0.5 to 1.5 parts of eggplant, 0.5 to 1.5 parts of turmeric, 0.5 to 1.5 parts of common monkshood mother root, 0.5 to 1.5 parts of kusnezoff monkshood root and 1 to 3 parts of Chinese azalea flower.
Preferably, the raw material medicines comprise the following components in parts by weight: 1 part of ephedra, 1 part of eggplant, 1 part of turmeric, 1 part of monkshood, 1 part of kusnezoff monkshood root and 2 parts of Chinese azalea flower.
Further, the medicine is an external medicine.
Furthermore, the medicine is an external preparation prepared by adding pharmaceutically acceptable auxiliary materials or auxiliary components into crude medicine powder, water or organic solvent extract of the raw medicine materials according to the weight ratio.
Further, the preparation is gel, paste, membrane, tincture, liniment, cataplasm or paste.
The invention provides an external pharmaceutical composition with anti-inflammatory and analgesic effects, which is a gel prepared from the following raw material medicines and auxiliary materials in parts by weight:
0.5 to 1.5 parts of ephedra, 0.5 to 1.5 parts of eggplant, 0.5 to 1.5 parts of turmeric, 0.5 to 1.5 parts of common monkshood mother root, 0.5 to 1.5 parts of kusnezoff monkshood root and 1 to 3 parts of Chinese azalea flower;
0.5-4 parts of sodium carboxymethylcellulose, 1-8 parts of propylene glycol and 1-8 parts of water.
Preferably, the external pharmaceutical composition is a gel prepared from the following raw material medicines and auxiliary materials in parts by weight:
1 part of ephedra, 1 part of eggplant, 1 part of turmeric, 1 part of monkshood, 1 part of kusnezoff monkshood root and 2 parts of Chinese azalea flower;
2.5 parts of sodium carboxymethylcellulose, 5 parts of propylene glycol and 5 parts of water.
The invention provides a preparation method of the external pharmaceutical composition, which comprises the following steps:
a. weighing the raw material medicines according to the weight ratio, adding water or an organic solvent for extraction, and obtaining an extract for later use;
b. adding water into the extract obtained in the step a, and uniformly mixing for later use;
c. and c, uniformly grinding the sodium carboxymethylcellulose and the propylene glycol in a mortar, adding the solution obtained in the step b while grinding, swelling and uniformly grinding to obtain the gel.
Wherein the water extract is prepared by the following method: taking the raw materials according to the weight ratio, adding 12-15 parts of water each time, decocting for 2 times, 2 hours for the first time and 1.5 hours for the second time, collecting water decoction, and concentrating to obtain the traditional Chinese medicine.
Wherein the organic solvent extract is an ethanol extract, and the extraction method comprises the following steps: taking the raw material medicines according to the weight ratio, adding 60% v/v ethanol water solution for extraction, extracting for 2 times at 80-90 ℃ according to the material-liquid ratio of 1:6, extracting for 2 hours for the first time and extracting for 1.5 hours for the second time, collecting the extracting solution, and concentrating to obtain the traditional Chinese medicine composition.
The invention provides the application of the raw material medicines in the weight ratio in preparing anti-inflammatory and analgesic medicines, and further provides a gel prepared by taking the raw material medicines as active ingredients, so that the anti-inflammatory and analgesic effects of the gel can be fully exerted. The pharmacodynamic experiment shows that the raw material medicines in the weight ratio have obvious anti-inflammatory and analgesic effects, the effect is superior to that of a commercial medicine, namely nataline, the medicine has no obvious skin irritation, the safety is good, and the medicine has wide clinical application prospects.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Detailed Description
The raw materials and equipment used in the embodiment of the present invention are known products and obtained by purchasing commercially available products.
Example 1 preparation of aqueous extract of osteopontic hemp
Soaking herba Ephedrae 50g, fructus Solani Melongenae 50g, Curcuma rhizome 50g, radix Aconiti Kusnezoffii 50g, and flos Rhododendri mollis 100g in water for 0.5 hr, decocting for 2 times, adding 5250mL of water for the first time, and decocting for 2 hr; adding 4200mL of water for the second time, and decocting for 1.5 h. Filtering the water decoction, mixing filtrates, concentrating, drying, pulverizing with small pulverizer to obtain dry extract 50.9116g, placing in sealed bag, and drying in a desiccator.
Example 2 preparation of an alcoholic extract of an orthopaedic anesthetic
Taking 30g of ephedra, 30g of solanum torvum bunge, 30g of curcuma, 30g of radix aconiti agrestis and 60g of flos rhododendri mollis, adding 60% v/v ethanol water solution for extraction, wherein the material-liquid ratio is 1:6, extracting for 2 times at 80-90 ℃, extracting for 2 hours for the first time and 1.5 hours for the second time, filtering, combining filtrates, concentrating, drying, crushing by a small crusher to obtain 25.9415g of dry paste powder, placing in a sealed bag, and drying for later use.
EXAMPLE 3 preparation of gel of osteoplastic drugs according to the invention
Prescription: 1.224g of dry extract powder of whole bone and anaesthesia prepared according to example 2
Sodium carboxymethylcellulose (CMC-Na) 1.224g
Propylene glycol 15mL
15mL of distilled water
The preparation method comprises the following steps: and uniformly grinding CMC-Na and propylene glycol in a mortar, slowly adding the aqueous solution in which the bone-setting anesthetic alcohol-extracted dry powder is dissolved while grinding, and uniformly grinding after swelling to obtain the bone-setting anesthetic gel I.
EXAMPLE 4 preparation of gel of osteoplastic drugs according to the invention
Prescription: total bone anesthesia alcohol extract dry paste powder 2.448g prepared according to example 2
Sodium carboxymethylcellulose (CMC-Na) 2.448g
Propylene glycol 30mL
30mL of distilled water
The preparation method comprises the following steps: and uniformly grinding CMC-Na and propylene glycol in a mortar, slowly adding the aqueous solution in which the bone-setting anesthetic alcohol-extracted dry powder is dissolved while grinding, and uniformly grinding after swelling to obtain the bone-setting anesthetic gel II.
EXAMPLE 5 preparation of gel of osteoplastic drugs according to the invention
Prescription: total bone anesthesia alcohol extract dry paste powder 4.896g prepared according to example 2
Sodium carboxymethylcellulose (CMC-Na) 2.448g
Propylene glycol 32mL
Distilled water 20mL
The preparation method comprises the following steps: and uniformly grinding CMC-Na and propylene glycol in a mortar, slowly adding the aqueous solution in which the bone-setting anesthetic alcohol-extracted dry powder is dissolved while grinding, and uniformly grinding after swelling to obtain the bone-setting anesthetic gel III.
Comparative example 1 preparation of a gel blank matrix
Prescription: sodium carboxymethylcellulose (CMC-Na) 1.224g
Propylene glycol 15mL
15mL of distilled water
The preparation method comprises the following steps: firstly, uniformly grinding CMC-Na and propylene glycol in a mortar, then slowly adding distilled water while grinding, and uniformly grinding after swelling to obtain the blank matrix gel.
The beneficial effects of the invention are proved by the pharmacodynamic experiments below.
Experimental example 1 comparison of anti-inflammatory effects of aqueous extract and alcoholic extract prepared in examples 1 and 2
1 Material
1.1 animals SD rats 18, with body mass 250 + -10 g, half each female and half, provided by Shanghai Si Laike laboratory animals, Inc., laboratory animal license number: SCXK (Shanghai) 2012-0002.
1.2 aqueous extract of osteopontin (prepared according to example 1), alcoholic extract of osteopontin (prepared according to example 2).
1.3 reagent 1% carrageenan (SIGMA-ALDRICH, batch No.: 9000-07-1).
1.4 instruments ZH-YLS-7B toe volume measuring instrument (Anhui Zhenghua biological instruments, Inc.), AL204 type electronic balance (Metler-Tollido instruments, Shanghai, Inc.).
2 method
2.1 healthy SD rats were grouped into 18, male and female halves, which were randomly divided: the blank group, the external group for water extraction of the bone-setting anesthetic and the external group for alcohol extraction of the bone-setting anesthetic are 3 groups, and each group comprises 6.
2.2 modeling 0.1mL of 1% carrageenan was injected between the three and four toes of the right hind paw of each rat (the palm of the right hind paw was inserted into the ankle joint).
2.3 the external application group for applying the water extract of the osteopathy medicament is applied to the foot soles of the foot joints of rats according to the clinical oral equivalent dose (1.1025g crude drug/kg) of a human body; the external application group of the bone-setting narcotic alcohol is coated with the bone-setting narcotic alcohol extract according to the clinical oral equivalent dose (1.1025g crude drug/kg) of a human at the foot joint sole of a rat; the blank group was given 2 times daily for 3 days, with equal amounts of blank matrix (prepared according to comparative example 1) applied to the same site.
2.4 index detection 1h after last administration, marking the joint of the right hind paw, measuring the volume of the right hind paw of each rat 2 times by a paw volume measuring instrument, taking the average value as the volume of the normal paw before inflammation of each rat, injecting 0.1mL of carrageenan into the space between the three four toes of the right hind paw of each rat (the center of the paw of the right hind paw is inserted towards the ankle joint), simultaneously administering once more, taking the left paw as a normal control, measuring the volume of the paw of the rat at 0.67h, 2h and 4h after injection, respectively, taking the difference of the volume of the toes before and after inflammation as the swelling degree of the paw (the swelling degree is the average volume of the paw after inflammation-the average volume of the paw before inflammation), and calculating the inflammation inhibition rate (the inhibition rate is the average swelling degree of the blank group-the average swelling degree of the administration group)/the average swelling degree of the blank group).
2.5 statistical treatment of the experimental data
Figure BDA0001047355200000051
Indicate, adoptSPSS20.0 statistical software for one-way anova with a t-test for comparison between groups, p<A difference of 0.05 is statistically significant.
3 results, see table 1.
The experimental results are as follows: compared with the blank group, the external application of the water extract and the alcohol extract of the osteopontin can generate obvious inhibition effect on the swelling reaction of the feet of the rat caused by the carrageenan; wherein, the anti-inflammatory effect of the alcohol extract of the osteopontin is better than that of the water extract.
Experimental example 2 anti-inflammatory action of osteopontic
1 Material
1.1 animals SD rats 60, with body mass 250 + -10 g, half each female and half, provided by Shanghai Si Laike laboratory animals, Inc., laboratory animal license number: SCXK (Shanghai) 2012-0002.
1.2 osteopontic anesthetic gels I to III (prepared according to examples 3 to 5).
1.3 reagent 1% carrageenan (SIGMA-ALDRICH, batch No.: 9000-07-1).
1.4 instruments ZH-YLS-7B toe volume measuring instrument (Anhui Zhenghua biological instruments, Inc.), AL204 type electronic balance (Metler-Tollido instruments, Shanghai, Inc.).
2 method
2.1 healthy SD rats were grouped into 60, male and female halves, and randomly divided into: blank group, low, medium and high dosage groups of the osteopontic anesthetic gel, wherein the low, medium and high dosage groups are divided into three measurement groups of 0.67h, 2h and 4h, 10 groups are provided, and 6 are provided in each group.
2.2 modeling 0.1mL of 1% carrageenan was injected between the three and four toes of the right hind paw of each rat (the palm of the right hind paw was inserted into the ankle joint).
2.3 administering the osteopathy anesthetic gel the low, medium and high dose groups are respectively administered with osteopathy anesthetic gel I, II and III, the administration doses of the low, medium and high dose groups are respectively 1.1025g crude drug/kg, 2.205g crude drug/kg and 4.41g crude drug/kg according to the set dose (the low dose group is clinical oral equivalent dose, the medium and high dose groups are respectively 2 times and 4 times of clinical oral equivalent dose), the administration doses of the low, medium and high dose groups are respectively 1.1025g crude drug/kg, 2.205g crude drug/kg and 4.41g crude drug/kg), the blank group is applied with an equivalent blank matrix (prepared according to the comparative example 1); it is continued for 2 times at intervals of 30 min.
2.4 index detection 1h after last administration, marking the joint of the right hind paw, measuring the volume of the right hind paw of each rat 2 times by a paw volume measuring instrument, taking the average value as the volume of the normal paw before inflammation of each rat, injecting 0.1mL of carrageenan into the space between the three four toes of the right hind paw of each rat (the center of the paw of the right hind paw is inserted towards the ankle joint), simultaneously administering once more, taking the left paw as a normal control, measuring the volume of the paw of the rat at 0.67h, 2h and 4h after injection, respectively, taking the difference of the volume of the toes before and after inflammation as the swelling degree of the paw (the swelling degree is the average volume of the paw after inflammation-the average volume of the paw before inflammation), and calculating the inflammation inhibition rate (the inhibition rate is the average swelling degree of the blank group-the average swelling degree of the administration group)/the average swelling degree of the blank group).
2.5 statistical treatment of the experimental data
Figure BDA0001047355200000061
Showing that the SPSS20.0 statistical software is used for one-way anova, and the comparison among groups is performed by t test and p is used<A difference of 0.05 is statistically significant.
3 results, see table 2.
The experimental results are as follows: compared with the blank group, the low-dose group has a significant difference in the foot swelling degree only 2h after the administration (P < 0.05); the swelling degree of feet of the medium-dose group is significantly different at 2h and 4h after administration (P <0.05 or P < 0.01); the high-dose groups have significant difference in foot swelling degree at 0.67h, 2h and 4h after administration (P <0.05 or P < 0.01).
The results show that the osteopathy anesthetic has obvious inhibition effect on rat paw swelling reaction caused by carrageenan, and the anti-inflammatory effect of medium and high dose is better than that of low dose.
Experimental example 3 comparison of anti-inflammatory Effect of osteopontic anesthetic and Futaline
1 Material
1.1 animals SD rats 60, with body mass 250 + -10 g, half each female and half, provided by Shanghai Si Laike laboratory animals, Inc., laboratory animal license number: SCXK (Shanghai) 2012-0002.
1.2 medicinal osteopontic anesthetic gel (prepared according to examples 4, 5); futallin emulsion (Beijing Nowa pharmaceutical Co., Ltd., batch No. X5086).
1.3 reagent 1% carrageenan (SIGMA-ALDRICH, batch No.: 9000-07-1).
1.4 instruments ZH-YLS-7B toe volume measuring instrument (Anhui Zhenghua biological instruments, Inc.), AL204 type electronic balance (Metler-Tollido instruments, Shanghai, Inc.).
2 method
2.1 healthy SD rats were grouped into 60, male and female halves, and randomly divided into: blank group, hibirine group, medium and high dosage group of the osteopathy anesthetic gel, wherein the middle and high dosage groups of the hibirine group, the osteopathy anesthetic gel are divided into three measurement groups of 0.67h, 2h and 4h, 10 groups are provided, and each group comprises 6.
2.2 modeling 0.1mL of 1% carrageenan was injected between the three and four toes of the right hind paw of each rat (the palm of the right hind paw was inserted into the ankle joint).
2.3 administration of each group, the middle and high dose groups of the osteopontic anesthetic gel (the administration dose of the middle and high dose groups is 2.205g crude drug/kg and 4.41g crude drug/kg respectively) are coated with corresponding drugs on the soles of the foot joints of rats according to the set dose [ the hibirine group (0.5g/kg), the blank group is coated with the same amount of blank matrix (prepared according to the comparative example 1) continuously for 2 times, and each time is 30 min.
2.4 index detection 1h after last administration, marking the joint of the right hind paw, measuring the volume of the right hind paw of each rat 2 times by a paw volume measuring instrument, taking the average value as the volume of the normal paw before inflammation of each rat, injecting 0.1mL of carrageenan into the space between the three four toes of the right hind paw of each rat (the center of the paw of the right hind paw is inserted towards the ankle joint), simultaneously administering once more, taking the left paw as a normal control, measuring the volume of the paw of the rat at 0.67h, 2h and 4h after injection, respectively, taking the difference of the volume of the toes before and after inflammation as the swelling degree of the paw (the swelling degree is the average volume of the paw after inflammation-the average volume of the paw before inflammation), and calculating the inflammation inhibition rate (the inhibition rate is the average swelling degree of the blank group-the average swelling degree of the administration group)/the average swelling degree of the blank group).
2.5 statistical treatment of the number of experimentsAccording to all are provided with
Figure BDA0001047355200000071
Showing that the SPSS20.0 statistical software is used for one-way anova, and the comparison among groups is performed by t test and p is used<A difference of 0.05 is statistically significant.
3 results, see table 3.
The experimental results are as follows: compared with the blank group, the medium-dose group has significant difference (P <0.05 or P <0.01) in the foot swelling degrees at 2h and 4h after the administration, and the high-dose group has significant difference (P <0.05 or P <0.01) in the foot swelling degrees at 0.67h, 2h and 4h after the administration, which indicates that the anti-inflammatory effect of the high-dose group is slightly better than that of the medium-dose group. Compared with the hibitane control group, the swelling inhibition rates of the osteopontic anesthetic high-dose groups 2h and 4h are 66.7% and 57.44% respectively, and are respectively higher than 60.68% and 44.61% of the hibitane group.
The results of the experiments are combined, which shows that the osteopathy anesthetic has obvious anti-inflammatory effect and can reach the level equivalent to that of common anti-inflammatory drug, namely, sitagliptin.
Experimental example 4 Hot plate method was used to examine analgesic effect of osteopontic anesthetic
1 Material
1.1 animals SPF-class Kunming species mice 70, 4 weeks old, female, body mass 20 + -2 g, provided by Shanghai Si Laike laboratory animals, Inc., laboratory animal license number: SCXK (Shanghai) 2012-0002.
1.2 medicinal bone setting anesthetic gel (self-made in laboratories); futallin emulsion (Beijing Nowa pharmaceutical Co., Ltd., batch No. X5086).
1.3 instruments RB-200 Intelligent hotplate instrument (Chengtai alliance software Co., Ltd.), AL204 electronic balance (Mettler-Tollido instruments Shanghai Co., Ltd.).
2 method
The mice are placed on a hot plate instrument (55 +/-0.5) DEG C one by one, timing is carried out immediately, and the time(s) required by the mice from the placement on the hot plate to the reaction of feet after licking is recorded as the normal pain threshold value of the mice. Selecting mice with pain threshold within 5-30 s, and discarding the mice with time less than or greater than the time of adding feet and unstable jumping.
Taking 60 qualified mice, randomly dividing the mice into a blank group, a model group (blank matrix group), a positive control group (hibirine group), a low, medium and high bone-setting anesthetic gel agent dosage group, wherein each group comprises 10 mice, and smearing the drugs for 2 times by each group according to the set dosage (the equivalent dosage conversion ratio of the mice to human bodies is 12:1, the equivalent dosage conversion ratio of the hibirine group and the bone-setting anesthetic gel agent is 0.95g/kg, the equivalent dosage conversion ratio of the mice to the human bodies is 2.1g of crude drugs/kg, the equivalent dosage conversion ratio of the model group to the low, medium and high bone-setting anesthetic gel agent is 4.2g of crude drugs/kg, the equivalent dosage conversion ratio of the model group to the human bodies is 8.4g of crude drugs/kg, and smearing the blank matrix. Each group of mice is placed in a hot plate, the time from the mice to the occurrence of the reaction of licking the hind paw is recorded by using the licking hind paw as an index of the pain reaction, for example, the mice which do not react within 60s are immediately taken out, and the pain threshold value is calculated according to 60 s. The pain threshold value is measured after smearing and administration for 0.5h, 1h and 2h, and the pain threshold percentage is calculated.
Pain threshold increase rate (post-dose pain threshold-pre-dose pain threshold)/pre-dose pain threshold 100%
3 results, see table 4.
The experimental results are as follows: compared with the blank group, the pain threshold values of the low-dose group and the high-dose group of the osteopathy have significant difference (P <0.01) at 1h and 2h after the administration, and the pain threshold values of the medium-dose group and the high-dose group of the osteopathy have significant difference (P <0.01) at 0.5h, 1h and 2h after the administration. Compared with the blank matrix group, the pain threshold values of the low-dose group of the osteopathy anesthetic are significantly different (P is less than 0.01) at 2h after the administration, and the pain threshold values of the medium-dose group and the high-dose group of the osteopathy anesthetic are significantly different (P is less than 0.01) at 0.5h, 1h and 2h after the administration. Compared with a sitagliptin control group, the pain threshold values of the low-dose group and the medium-dose group are not significantly different at 0.5h, 1h and 2h after administration (P is more than 0.05); the pain threshold values of the high-dose groups of the osteopontic anesthetics at 1h and 2h after administration are both significantly different (P < 0.05).
The results show that the high dose of the osteopathic anesthetic has better inhibition effect on pain caused by mouse hot plate, and the curative effect is better than that of the sitagliptin.
Experimental example 5 analgesic action of osteopontic anesthetic was examined by acetic acid-induced writhing response experiment in mice
1 Material
1.1 animals SPF-level Kunming mice 60, 4 weeks old, female, body mass 20 + -2 g, provided by Shanghai Si Laike laboratory animals, Inc., laboratory animal license number: SCXK (Shanghai) 2012-0002.
1.2 medicinal bone setting anesthetic gel (self-made in laboratories); futalin emulsion (Beijing Nowa pharmaceutical Co., Ltd., batch No. X5086); acetic acid (national chemical group, Ltd., lot number: 20141215).
1.3 instruments RB-200 Intelligent hotplate instrument (Chengtai alliance software Co., Ltd.), AL204 electronic balance (Mettler-Tollido instruments Shanghai Co., Ltd.).
2 method
60 SPF mice are randomly divided into blank groups, model groups, positive control groups (hibirine groups), low, medium and high-dose groups of the osteopathy anesthetic, and 10 mice in each group, wherein each group is coated and administrated on the abdomen for 2 times according to set doses (hibirine groups, low, medium and high-dose groups of osteopathy anesthetic gel are respectively 0.95g/kg, 2.1g crude drugs/kg, 4.2g crude drugs/kg and 8.4g crude drugs/kg, the model groups are coated with equal amount of blank matrix), the drugs are coated for 1 time every 30min, and the blank groups are not treated. A0.6% acetic acid solution was injected into the abdominal cavity (0.2 mL/mouse) 1h after the last administration to induce a large area and long lasting painful irritation in the deep part, resulting in a writhing response in the mouse. The abdomen depression and the extension of the two hind limbs of the mouse are used as the index of the writhing response. The time (latency) for the writhing response of each group of mice and the number of writhing responses within 15min were observed.
3 results, see table 5.
The experimental results are as follows: compared with the blank group, the low-dose group of the osteopontin can prolong the latency period of writhing response of mice, but the difference is not statistically significant (P is more than 0.05); the frequency of writhing is reduced within 15min in the low dose group of the osteopathy and anesthesia medicine, and the difference has statistical significance (P is less than 0.05); the medium and high dose groups of the osteopontic anesthetic can obviously prolong the latency period of writhing response of mice (P <0.05 or P <0.01), and the writhing frequency is obviously reduced within 15min (P < 0.01). Compared with the blank matrix group, the low-dose group of the osteopontin can prolong the latency period of writhing response of mice, the times of writhing are reduced within 15min, but the difference is not statistically significant (P is more than 0.05); the medium and high dose groups of the osteopontic anesthetic can obviously prolong the latency period of writhing response of mice (P <0.05 or P <0.01), and the number of writhing times is obviously reduced within 15min (P <0.05 or P < 0.01). Compared with a hibitane control group, the middle and high dose groups of the osteopontic anesthetics can prolong the latency period of writhing response of mice, the number of writhing times is reduced within 15min, but the difference has no statistical significance (P is more than 0.05).
The results of the experiments show that the bone-setting anesthetic can play a significant analgesic role by external application, and the effect is superior to or equivalent to that of sitagliptin.
Experimental example 6 Experimental study on skin irritation of osteoplastic drugs
1. Material
1.1 animals SPF-level Kunming species mice 20, 4 weeks old, half female and half male, body mass 20 + -2 g, provided by Shanghai Si Laike laboratory animals, Inc., laboratory animal license number: SCXK (Shanghai) 2012-0002.
1.2 pharmaceutical osteopontin gel (prepared according to example 5); excipient: vaseline (Jiangxi division of medical science and technology, Inc., Shandong Rierkang, batch No. 150529).
2. Method of producing a composite material
2.1 animals 20 mice were divided into groups and randomized: intact skin test agent, vehicle group and damaged skin test agent, vehicle group, total 2 groups, each group 10.
2.2 test method ① setting excipient control, using the same body left and right self-contrast method, i.e. coating vaseline on the left side depilatory area, coating whole bone anesthesia gel on the right side depilatory area, depilating treatment on the administration area (both sides of back vertebral column) 24h before ② administration, checking whether the depilated skin is damaged before administration of 3cm × 3 cm. on the left and right sides of depilatory area, and the damaged skin should not be tested completely, when researching the damaged skin, cutting a 'well' on the depilatory area with a blade, and using the degree of blood seepage, ③ whole bone anesthesia gel is coated on the right depilatory area, coating vaseline on the left depilatory area, then covering with two layers of gauze and a layer of cellophane or the like, fixing with medical rubber plaster, after ④ 24h, washing out warm water whole bone anesthesia gel, removing gel after removing the test substance, observing whether there is erythema and edema reaction on the coated area 1, 24, 48, 72h, ⑤ test results are scored standard tables 6 and 7.
3 results, see Table 8
The experimental results are as follows: the skin of mice in the intact skin excipient control group and the damaged skin excipient control group has no skin irritation reaction such as erythema, edema and the like, and the skin of the administration part of the osteopathy and anesthesia has no other abnormal reaction. The general state, behavior, physical signs and the like of the animals in each group are not abnormal.
The results show that the bone-setting anesthetic has no irritation to skin and good safety.
Figure BDA0001047355200000111
Figure BDA0001047355200000131
Figure BDA0001047355200000151
Figure BDA0001047355200000161

Claims (7)

1. The application of the following raw material medicines in preparing the anti-inflammatory and detumescent medicine in weight ratio is as follows: 0.5 to 1.5 parts of ephedra, 0.5 to 1.5 parts of eggplant, 0.5 to 1.5 parts of turmeric, 0.5 to 1.5 parts of common monkshood mother root, 0.5 to 1.5 parts of kusnezoff monkshood root and 1 to 3 parts of Chinese azalea flower; the medicine is an external medicine and is an external preparation prepared by adding pharmaceutically acceptable auxiliary materials or auxiliary components into crude medicine powder, water or organic solvent extract of raw medicine materials according to the weight ratio; the external preparation is gel, paste, membrane, tincture, liniment, cataplasm or cataplasm.
2. Use according to claim 1, characterized in that: the weight ratio of the raw material medicines is as follows: 1 part of ephedra, 1 part of eggplant, 1 part of turmeric, 1 part of monkshood, 1 part of kusnezoff monkshood root and 2 parts of Chinese azalea flower.
3. Use according to claim 1, characterized in that: the gel is prepared from the following raw material medicines and auxiliary materials in parts by weight:
0.5 to 1.5 parts of ephedra, 0.5 to 1.5 parts of eggplant, 0.5 to 1.5 parts of turmeric, 0.5 to 1.5 parts of common monkshood mother root, 0.5 to 1.5 parts of kusnezoff monkshood root and 1 to 3 parts of Chinese azalea flower;
0.5-4 parts of sodium carboxymethylcellulose, 1-8 parts of propylene glycol and 1-8 parts of water.
4. Use according to claim 3, characterized in that: the gel is prepared from the following raw material medicines and auxiliary materials in parts by weight:
1 part of ephedra, 1 part of eggplant, 1 part of turmeric, 1 part of monkshood, 1 part of kusnezoff monkshood root and 2 parts of Chinese azalea flower; 2.5 parts of sodium carboxymethylcellulose, 5 parts of propylene glycol and 5 parts of water.
5. Use according to claim 4, characterized in that: the preparation method of the gel comprises the following steps:
a. weighing the raw material medicines according to the weight ratio, adding water or an organic solvent for extraction, and obtaining an extract for later use;
b. adding water into the extract obtained in the step a, and uniformly mixing for later use;
c. and c, uniformly grinding the sodium carboxymethylcellulose and the propylene glycol in a mortar, adding the solution obtained in the step b while grinding, swelling and uniformly grinding to obtain the gel.
6. Use according to claim 1, characterized in that: the water extract is prepared by the following method: taking the raw materials according to the weight ratio, adding 12-15 parts of water each time, decocting for 2 times, 2 hours for the first time and 1.5 hours for the second time, collecting water decoction, and concentrating to obtain the traditional Chinese medicine.
7. Use according to claim 1, characterized in that: the organic solvent extract is an ethanol extract, and the extraction method comprises the following steps: taking the raw material medicines according to the weight ratio, adding 60% v/v ethanol water solution for extraction, extracting for 2 times at 80-90 ℃ according to the material-liquid ratio of 1:6, extracting for 2 hours for the first time and extracting for 1.5 hours for the second time, collecting the extracting solution, and concentrating to obtain the traditional Chinese medicine composition.
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Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
"祖国医学关于麻醉药的文献";余尚德;《上海中医药杂志1956年5月号》;19560531;28-29 *
"祖国医学关于麻醉药的文献";余尚德;《上海中医药杂志1956年5月号》;19560531;575 *
"闹羊花治疗类风湿性关节炎258例疗效观察";孙健 等;《新医学1989年第20卷第11期》;19891231;28-29 *

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