CN113274449A - Nine-ingredient gardenia red liniment for treating acute soft tissue injury and preparation method thereof - Google Patents
Nine-ingredient gardenia red liniment for treating acute soft tissue injury and preparation method thereof Download PDFInfo
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- CN113274449A CN113274449A CN202110704687.7A CN202110704687A CN113274449A CN 113274449 A CN113274449 A CN 113274449A CN 202110704687 A CN202110704687 A CN 202110704687A CN 113274449 A CN113274449 A CN 113274449A
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Abstract
The invention discloses a nine-ingredient gardenia red liniment for treating acute soft tissue injury and a preparation method thereof, and relates to the technical field of medicines. The nine-ingredient gardenia red liniment comprises: 700-1300 parts of cape jasmine, 700-1300 parts of safflower, 350-650 parts of clematis root, 350-650 parts of angelica, 210-390 parts of ground beetle, 210-390 parts of borneol, 70-130 parts of frankincense, 70-130 parts of myrrh and 70-130 parts of dragon's blood. The preparation method comprises the following steps: weighing medicinal materials except borneol according to the prescription proportion, uniformly mixing, crushing, sieving, soaking for 48 hours by using 4-6 times of 70-90% ethanol in volume according to the proportion of 1g of the medicinal materials to 1ml of solvent, slowly percolating, collecting initial percolate for later use, then slowly percolating by adding 4-6 times of 70-90% ethanol in volume, concentrating the percolate under reduced pressure until no alcohol smell exists, combining the percolate with the initial percolate, adding crushed borneol according to the prescription proportion, adding a proper amount of 95% ethanol to uniformly disperse, then adding pure water to adjust the total amount of the solution to be 6.21-6.61 times of the total amount of the medicinal materials in volume of ethanol solution, shaking uniformly, and subpackaging to obtain the borneol-mixed with the total amount. Has better detumescence and pain-relieving effects on treating acute soft tissue injury.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to application of nine-ingredient gardenia red liniment in preparation of medicines for treating acute soft tissue injury.
Background
Acute soft tissue injury (acute soft tissue injury) is a common clinical orthopedic injury, and generally refers to closed injury of muscles, tendons, ligaments and the like of a human body caused by falling, pounding, flashing, twisting, bumping and the like [ Cao Qing, Shen, Yan Lian, and the like. 244-246.]. Common soft tissue injuries such as skin, subcutaneous tissue, fascia, muscle, tendon ligament, periosteum, joint capsule and the like are caused, and peripheral nerve vessel injuries are combined; the main symptoms are manifested by bluish purple ecchymoses, swelling and pain of affected parts of patients, or accompanied by dysfunction [ Zhang Gui Fu, Wang Shi Yi, research progress of action mechanism of traditional Chinese medicine for treating soft tissue injury, science and technology of traditional Chinese medicine, 2011, 18 (2): 174-175.]. Generally, the acute soft tissue injury of a patient has no serious consequence, but also affects the work, study and life of the patient to a certain extent. The key to acute soft tissue injury treatment lies in pain relief and spasm relief [ Zhou Ben Cheng, Shen Jiang, wrist and ankle needle combined with acupuncture "Ashi point" and body needle treatment for acute soft tissue injury 166 cases, Zhejiang journal of traditional Chinese medicine, 2008, 43 (8): 469-470.]. Modern medicine mainly adopts methods such as early braking, bandaging, local sealing, medicines, physical therapy and the like for treatment, generally oral or external antipyretic analgesics and non-steroidal anti-inflammatory drugs are used for controlling symptoms, and hormones are added in a short time when the symptoms are serious. The above drugs are often not good in curative effect in the using process, and are easily accompanied with adverse reactions [ McGettigan P, Henry d. current schemes with non-specific Cox inhibitors. current pharmaceutical design, 2000, 6 (17): 1693-1724 Andres BM, Murrefi GAC. treatment of tendinopathy, what works, what doss not, and what is on the horizons, clinical Orthopaedics and Related Research, 2008, 466 (7): 1539 1554. severe organ damage can even occur [ Wright HN. viral toxicity students of acquired and antigenic drugs and regulators. toxicol and Applied Pharmacology, 1967, 11 (2): 280-292, heartray WFW.hematology effects of antigenic industries, drug-induced aggregations.American Journal of Medicine, 1983, 75 (5): sharma S, Prasad A, Anand KS. nonsteroidal anti-reflective drugs in the management of a pad and an reflective: A basis for a drug selection, American Journal of Therapeutics, 1999, 6(1):3., 1999, 6(1): hawkins C, Hanks GW, the structural sensitivity of non-organic anti-reflective drugs, A review of the performance, journal of paint and Symptom Management, 2000, 20 (2): 140-151.].
From ancient times to present, traditional Chinese medicine has abundant clinical practice experience in the aspect of treating acute soft tissue injury, and increasingly attracts attention of medical workers due to unique mechanism and good curative effect. The traditional Chinese medicine theory considers that acute soft tissue injury belongs to the category of 'tendon injury', is tendon and muscle injury caused by external violence acting on a human body, and mainly comprises qi stagnation, blood stasis and vein obstruction [ Shanzhong school, Gardenia red rhubarb liniment treats 121 cases of soft tissue injury, Shanxi Chinese medicine, 1994, 15 (6): 256 of Lujianhua, Yanhuarui, Liuwanjun, and the like, the experimental study on the external application of gardenia honey paste for treating the acute soft tissue injury of rabbits, the journal of orthopedics and traumatology in Chinese, 2017, 25 (8): 6-10.]. The book of the theory of yin-yang correspondence in Huangdi's internal classic & Su Wen (yellow emperor's classic of things) is in large term: "Qi impairment pain, form impairment and swelling". The general prescription and the door of fracture are recorded in the book: "if the blood circulation is obstructed due to injury and rupture, internal movement of tendons and collaterals, and fatigue and accumulation are not dispersed, it is considered as swelling and pain, and it is advisable to remove fatigue and circulate qi and blood. It is considered that "qi stagnation and blood stasis and vein obstruction" occur during injury of tendons and tendons, blood stasis occurs due to qi stagnation, and blood stasis causes qi stagnation, thereby causing blood to be retained on the surface of skin, and the skin to present gall and bluish purple [ Yuan Di, Zhu Shao, Shenyan, etc.. the clinical research progress of the external treatment of acute soft tissue injury in the traditional Chinese medicine, journal of external treatment in the traditional Chinese medicine, 2008, 17 (1): 41-43.].
In recent years, with the continuous and deep development of the research on the mechanism of treating soft tissue injury in traditional Chinese medicine, the action mechanism of the medicine is continuously discovered, and a solid theoretical basis is provided for clinical application. After traumatic injury occurs, qi stagnation and blood stasis, vein blockage and blockage caused by blockage of local meridians and unsmooth blood circulation cause pain [ Cao Qing, Shen Lin, Yan Ping Zi Cao, etc.. the peril III clinical research of the peril-staltic-spur-bone spray for treating acute soft tissue injury (qi stagnation and blood stasis syndrome). the combined research of traditional Chinese and western medicines, 2009, 1 (5): 244, 246, leaf of Chaohui brightness, discussing the treatment of sinew injury in yellow emperor's internal classic, Henan TCM, 2013, 33 (10): 1620-1623.]. Severe pain, local swelling and dysfunction are the major clinical symptoms of acute soft tissue injury [ both, song together. People health press, 1990: 59-61.]. The traditional Chinese medicine mainly uses medicines with the functions of promoting blood circulation to remove blood stasis, promoting qi circulation to relieve pain, clearing heat and cooling blood and the like for treating the soft tissue injury [ zhaolili, chenweiliang ] traditional Chinese medicine external treatment acute soft tissue injury research near condition shallow analysis [ J ]. Chinese traditional medicine orthopedics journal, 2007, 15 (3): 60-61 Xuebin, xi Xiao Bing, Li Fei, clinical application progress of external application medicine for treating soft tissue injury [ J ]. Chinese J of orthopedics, 2014, 22 (6): 78-80.]. For example, cold compress applied at the early stage of injury can cause the blood vessels at the injury to contract, reduce bleeding and relieve pain; the treatment in the convalescent period is mainly the topical application of the medicine for relieving pain, promoting blood circulation and removing blood stasis; the blood circulation promoting and blood stasis removing medicine can relieve inflammatory reaction, promote tissue exudate absorption, improve pain threshold, and relieve pain.
Therefore, a Chinese medicinal compound external preparation, namely a nine-ingredient gardenia red liniment, is researched and prepared by penmen and is used for treating acute soft tissue injury.
Disclosure of Invention
The invention provides a nine-ingredient gardenia red liniment for treating acute soft tissue injury and a preparation method thereof.
The nine-ingredient gardenia red liniment is prepared from nine medicinal materials, namely gardenia, safflower, clematis root, Chinese angelica, ground beeltle, borneol, frankincense, myrrh and dragon's blood, according to the following parts by mass: 700-1300 parts of cape jasmine, 700-1300 parts of safflower, 350-650 parts of clematis root, 350-650 parts of angelica, 210-390 parts of ground beetle, 210-390 parts of borneol, 70-130 parts of frankincense, 70-130 parts of myrrh and 70-130 parts of dragon's blood.
Preferably, the composition is prepared from the following components in parts by mass: 800-1200 parts of cape jasmine, 800-1200 parts of safflower, 400-600 parts of radix clematidis, 400-600 parts of angelica sinensis, 240-360 parts of ground beetle, 240-360 parts of borneol, 80-120 parts of frankincense, 80-120 parts of myrrh and 80-120 parts of dragon's blood.
Further preferably, the composition is prepared from the following components in parts by mass: 900-1100 parts of cape jasmine, 900-1100 parts of safflower, 450-550 parts of clematis root, 450-550 parts of angelica, 270-330 parts of ground beetle, 270-330 parts of borneol, 90-110 parts of frankincense, 90-110 parts of myrrh and 90-110 parts of dragon's blood.
The nine-ingredient gardenia red liniment is prepared by the following preparation method: weighing the medicinal materials of gardenia, safflower, clematis root, angelica, ground beeltle, frankincense, myrrh and dragon blood according to the prescription proportion, uniformly mixing, crushing and sieving; soaking 1g of each medicinal material in 70-90% ethanol with the volume of 4-6 times of that of 1g of the medicinal material to 1ml of a solvent for 48 hours, slowly percolating, collecting initial percolate for later use, adding 70-90% ethanol with the volume of 4-6 times of that of the medicinal material to slowly percolating, concentrating the percolate under reduced pressure until no alcohol smell exists, combining the percolate with the initial percolate, adding borneol with the crushed formula amount proportion, adding a proper amount of 95% ethanol to uniformly disperse, adding pure water to adjust the total amount of the solution to be 6.21-6.61 times of that of the total amount of the medicinal materials, shaking uniformly, and subpackaging to obtain the traditional Chinese medicine.
Further preferably, the nine-ingredient gardenia red liniment is prepared by the following preparation method: weighing gardenia, safflower, clematis root, angelica, ground beeltle, frankincense, myrrh and dragon's blood according to the prescription proportion, uniformly mixing and crushing the materials, wherein each part of the materials is 1g, sieving the materials by a 80-mesh sieve, soaking the materials by using 5 times volume of 80% ethanol for 48h according to the proportion of 1g of the materials to 1ml of solvent, slowly percolating the materials, collecting initial percolate for later use, then adding 5 times volume of 80% ethanol for slow percolation, decompressing and concentrating the percolate until no ethanol smell exists, merging the percolate with the initial percolate, adding crushed borneol according to the prescription proportion, adding 95% ethanol with the volume of 0.17 time volume of the total amount of the materials for uniform dispersion, adding pure water to adjust the total amount of the solution to be 6.41 times volume of ethanol solution of the total amount of the materials, shaking the solution uniformly, and subpackaging the mixture with the density of 0.90 g/ml-0.94 g/ml to obtain the traditional Chinese medicine.
The application effect of the nine-ingredient gardenia red liniment in preparing the medicine for treating acute soft tissue injury is verified through the following animal experiments and clinical trials:
1. animal experiments: selecting male Kunming mice, randomly dividing into 4 groups, respectively externally coating nine gardenia red liniments, safflower oil, 60% ethanol solution and normal saline, and inspecting the influence of the nine gardenia red liniments on the increase of the permeability of capillary vessels in the abdominal cavity of the mice caused by acetic acid through the concentration of Evans blue in the recovered abdominal cavity liquid. Selecting male big-ear white rabbits, randomly dividing the male big-ear white rabbits into 4 groups, manufacturing an acute soft tissue injury animal model, respectively externally coating nine gardenia red liniments, safflower oil, 60% ethanol solution and normal saline for treatment, detecting the plasma viscosity through peripheral blood, measuring the content of interleukin-1 beta through homogenate of muscle tissues of an impacted part, and inspecting the influence of the nine gardenia red liniments on the plasma viscosity of the acute soft tissue injury rabbits and the content of the interleukin-1 beta of tissues of the injured part. The results show that the nine-ingredient gardenia red liniment can obviously inhibit the increase of the permeability of the capillary vessels in the abdominal cavity of a mouse caused by acetic acid, obviously reduce the blood plasma viscosity of rabbits with acute soft tissue injury, obviously reduce the IL-1 beta content of local tissues and has good effects of promoting blood circulation and removing blood stasis.
2. And (3) clinical trials: selecting 2 groups of patients with acute soft tissue injury after exercise, and respectively externally coating nine gardenia red liniments and safflower oil within 24-48 h after injury for 2 times/day with the dosage of 0.5ml/cm each time2The clinical efficacy of the nine-ingredient gardenia red liniment is evaluated by observing the change conditions of blood stasis, swelling and pain symptoms at the soft tissue injury part before and after treatment. The results show that the nine-ingredient gardenia red liniment can obviously improve the swelling and pain of the acute soft tissue injury of the patient, has better treatment effect on the acute soft tissue injury, and is an ideal medicine for treating the acute soft tissue injury.
The nine-ingredient gardenia red liniment has the following advantages:
1. the nine-ingredient gardenia red liniment is derived from a traditional Chinese medicine classic formula gardenia red flower medicine pair, clematis root, Chinese angelica, ground beeltle, borneol, frankincense, myrrh and dragon blood are added according to the syndrome differentiation, the prescription is composed of nine medicinal materials of gardenia, safflower, clematis root, Chinese angelica, ground beeltle, borneol, frankincense, myrrh and dragon blood, and all the medicinal materials are nontoxic and harmless, so that the nine-ingredient gardenia red liniment is a pure natural Chinese medicine compound external preparation.
2. The nine-ingredient gardenia red liniment conforms to the formula principle of the traditional Chinese medicine, the pair of gardenia red anther is a good medicine for treating tendon injury, namely acute soft tissue injury in the traditional Chinese medicine, gardenia has the functions of clearing heat and promoting diuresis, cooling blood and removing toxicity, and relieving swelling and pain, and safflower has the functions of activating blood and stimulating the menstrual flow, dissipating blood stasis and relieving pain, and inducing diuresis to alleviate edema; the other traditional Chinese medicines comprise clematis root, radix clematidis, clematis root, borneol, myrrh, frankincense and myrrh, wherein the clematis root is used for dispelling wind and removing dampness, dredging collaterals and relieving pain, the angelica is used for replenishing blood and activating blood circulation, removing blood stasis and relieving swelling, regulating menstruation and relieving pain, the ground beeltle is used for breaking blood stasis and reuniting muscles and bones, the borneol is used for inducing resuscitation and inducing resuscitation, clearing heat and relieving pain, the dragon's blood is used for activating blood and removing stasis, and promoting granulation and healing wound, the frankincense and the myrrh are essential medicines for activating blood and resolving hard mass, and the frankincense and the myrrh can be used for relieving pain, relieving swelling and promoting granulation in the cloud of Ben Cao gang mu, so that the two medicines are used together; the medicines are combined to be used for clearing away heat and toxic materials, promoting qi and blood circulation, removing blood stasis and swelling, and dredging collaterals and relieving pain.
3. The nine-ingredient gardenia red liniment highlights the basic characteristics of the combination of syndrome differentiation, etiological treatment and symptomatic treatment of the traditional Chinese medicine, can obviously reduce capillary permeability, reduce plasma viscosity and reduce the IL-1 beta content of local tissues aiming at the 'tendon injury' symptoms of qi stagnation and blood stasis and obstruction of veins during acute soft tissue injury and the main clinical manifestations of pain and swelling, has good effects of activating blood circulation to dissipate blood stasis, obviously improves the swelling and pain of the acute soft tissue injury of a patient, effectively reduces swelling and relieves pain, and has better treatment effect on the acute soft tissue injury.
4. The nine-ingredient gardenia red liniment is mainly used for treating acute soft tissue injury, adopts an alcohol-containing liquid liniment form, is convenient for patients to use, easy to absorb, quick in effect, convenient to carry and store, high in safety of external preparations, and has no toxic or side effect when used under normal dosage.
Detailed Description
The present invention is further illustrated by the following examples, but the practice of the invention is not limited thereto.
Example 1: preparation of nine-ingredient gardenia red liniment
Weighing 1000g of cape jasmine, 1000g of safflower, 500g of clematis root, 500g of angelica, 300g of ground beeltle, 100g of frankincense, 100g of myrrh and 100g of dragon's blood according to the prescription proportion of the nine-ingredient gardenia red liniment, uniformly mixing the medicinal materials, crushing the medicinal materials, sieving the medicinal materials by a 80-mesh sieve, soaking the medicinal materials by using 80% ethanol with the volume 5 times of the medicinal materials for 48 hours according to the proportion of 1g of the medicinal materials to 1ml of solvent, slowly percolating the medicinal materials, collecting initial percolate for later use, then adding 80% ethanol with the volume 5 times of the medicinal materials for slow percolation, concentrating the percolate under reduced pressure until no alcohol smell exists, merging the percolate with the initial percolate, adding 300g of crushed borneol, adding 650ml of 95% ethanol for uniform dispersion, then adding pure water to adjust the total amount of the solution to 25000ml, shaking the mixture evenly, measuring the split charging density to be 0.917g/ml, and filling the mixture in a bottle with 30 ml/bottle to obtain the gardenia red liniment.
Example 2: influence of nine-ingredient gardenia red liniment on permeability of capillary vessels in abdominal cavity of mouse
1 materials and methods
1.1 instruments and reagents
Electronic balance, UTP-313 type (maximum load 2000g, index 0.01g), product of Shanghai Ficko tools, Inc. A medical low speed centrifuge, model B600B, product of beijing white ocean centrifuge ltd. Ultraviolet spectrophotometer model 752N, Shanghai apparatus electric analyzer product, Inc. The water is double distilled water, and the ethanol and other reagents are analytically pure.
1.2 reagent
Jiuweizhihong liniment (Jiuweizhihong liniment) provided by people hospital pharmacy department of Taixing city, with specification of 30 ml/bottle and batch number 181202. Regular safflower oil (red flower oil), available from Wanyuan (Fuzhou) pharmaceutical Co., Ltd., 20 ml/bottle, lot number 110906. 60% ethanol solution (60% alcohol solution), prepared in pharmacy department of people hospital in Thxing City. 0.9% physiological saline solution (0.9% saline solution), 250 ml/bag, lot No. 09072884, manufactured by Erodies Geranii, Anhui, Co., Ltd.
1.3 Experimental animals
40 Kunming mice are male, have the body mass of 18 g-22 g and are clean, are provided by the experimental animal center of Xuzhou medical university, are bred in the animal room of the experimental center of hospital (the breeding temperature is 19-24 ℃, the relative humidity is 50-60 percent and the ventilation is good) with the production license number SCXK (Su) 2010-0003 and the use license number SYXK (Su) 2010-0011, and are fed with standard feed, are drunk freely and are fed adaptively for 1 week before experiments.
1.4 Experimental methods
Reference is made to the literature [ Blackham A, Woods FAM. Immune cMultiplex media information in the motor property of A method for inducing inhibition of metabolism of arachis salt method journal of Pharmacological Methods, 1986, 15 (1): 77-85. Liu loop, Chenglifang, Liu Yongzhi, and the like, research on analgesic and anti-inflammatory effects of hyperosteogeny analgesic plaster, college of Liaoning traditional Chinese medicine, 2009, 11 (12): 197-198.]The method is used for carrying out experiments for influencing the permeability of the capillary vessels in the abdominal cavity. 40 Kunming mice are randomly divided into 4 groups, namely nine gardenia red liniment groups, safflower oil groups, ethanol solution groups and normal saline control groups, wherein each group comprises 10 mice. Each group of mice had their abdomen cut into hairs with an exposed area of about 2cm2The nine-ingredient gardenia red liniment, the safflower oil, the 60 percent ethanol solution and the normal saline solution are respectively and correspondingly coated, the dosage is 0.8ml/kg for each time and 1 time/day for 3 days. After 1h on day 3, mice in each group were injected intravenously with 10ml/kg caudate 0.5% Evans blue (evans blue) solution followed by intraperitoneal injection of 10ml/kg 0.6% acetic acid solution. After 20min, the cervical vertebrae were removed and the groups of mice were sacrificed. Cutting the skin of the abdomen of the mouse, injecting 5ml of physiological saline for washing, gently kneading the abdomen of the mouse for 1min, sucking the washing liquid in the abdominal cavity by using a syringe, injecting the washing liquid into a 10ml centrifugal tube, and centrifuging the tube in a centrifuge at 3000r/min for 15 min. Taking 2ml of the supernatant, placing the supernatant in a sample pool of an ultraviolet spectrophotometer for colorimetric determination at 590nm, recording the OD value of absorbance, calculating the concentration of evans blue in the peritoneal liquid, and judging the influence of the nine-ingredient gardenia red liniment on the increase of the permeability of the capillary vessels in the peritoneal cavity of the mouse caused by acetic acid.
1.5 statistical treatment
The experimental results are mean ± standard deviation
Representing statistical treatment using SPSS 21.0 software, comparison of differences between groups using one-way analysis of variance, pairwise comparison between groups by LSD-t test, and P<0.05 is statistically significant.
2 results
On the 3 rd day after treatment, the recovery concentration of evans blue in the abdominal liquid of the mice of the nine-ingredient gardenia red liniment group is respectively and obviously lower than that of a normal saline control group and that of an ethanol solution group (P is less than 0.05); the recovery concentration of evans blue of the nine gardenia red liniment groups is lower than that of safflower oil, but the difference between the two groups has no statistical significance (P is more than 0.05). The results show that the nine-ingredient gardenia red liniment has the effect of remarkably reducing the recovery of evans blue in the abdominal liquid of the mice, namely the nine-ingredient gardenia red liniment can remarkably inhibit the increase of capillary permeability of the abdominal cavity of the mice caused by acetic acid, and the results are shown in table 1.
TABLE 1 results of measurements of the effect of increased permeability of capillary vessels in the abdominal cavity of mice caused by acetic acid
Note: compared with the normal saline control group,*P<0.05; compared with the ethanol solution group, the method has the advantages that,#P<0.05
3 conclusion
The nine-ingredient gardenia red liniment can obviously reduce the permeability of capillary vessels in the abdominal cavity of a mouse. After acute soft tissue injury occurs, the nine-ingredient gardenia red liniment is continuously applied for treatment in the early stage and can obviously inhibit the liquid exudation in capillary vessels at the injury part.
Example 3: influence of fructus Gardeniae red liniment on acute soft tissue injury rabbit plasma viscosity and local tissue interleukin-1 beta content
1 materials and methods
1.1 instruments and reagents
Electronic analytical balance, Quintix 224-1CN type (maximum load 220g, index 0.1mg), product of Sartorius, Germany. Electronic balance, UTP-313 type (maximum load 2000g, index 0.01g), product of Shanghai Ficko tools, Inc. A medical low speed centrifuge, model B600B, product of beijing white ocean centrifuge ltd. A full-automatic blood rheology rapid measuring instrument, FASCO-3010D type, product of Chongqing Victoria technology, Inc. Iodine [ 2 ]125I]Interleukin-1 beta (interleukin-1 beta, IL-1 beta) radioimmunoassay kit, product of research institute of radioimmunoassay of general Hospital science and technology development center of the people's liberation force, China, batch No. 090630. The water is double distilled water, and the ethanol and other reagents are analytically pure.
1.2 reagent
A Jiuwei Zhihong liniment, which is provided by people hospital pharmacy in Taixing city, with the specification of 30 ml/bottle and the batch number of 181202. Safflower oil, available from Wanyuan (Fuzhou) pharmaceutical Co., Ltd., 20 ml/bottle, lot No. 110906. 60% ethanol solution, prepared by pharmacy department of people hospital in Taixing city. 0.9% physiological saline solution, 250 ml/bag, lot No. 09072884, manufactured by Anhui double Crane pharmaceuticals Co. Heparin sodium injection (heparin sodium injection), 1.25 wu/count, lot number 1001115, manufactured by Jiangsu Wanbang Biochemical medicine group, Inc.
1.3 Experimental animals
20 big-ear white rabbits are male, the body mass is 2.0 kg-2.5 kg, the common level is provided by the experimental animal center of Xuzhou medical university, the production license number SCXK (Su) 2010-containing 0003 of the experimental animal and the use license number SYXK (Su) 2010-containing 0011 of the experimental animal are raised in the animal room of the experimental center of hospital (the raising temperature is 19-24 ℃, the relative humidity is 50-60%, the ventilation is good), the standard feed is adopted for feeding, the water is freely drunk, and the adaptive feeding is carried out for 1 week before the experiment.
1.4 Experimental methods
1.4.1 animal Molding
Reference is made to the literature [ cuulou, zhuyiping, kaiweijuan, etc.. experimental studies on the effect of ankle comforter on acute soft tissue injury PGE-2. traditional chinese medicine bonesetting, 2004, 16 (1): 15-17, zhangchen, wangho, daubing, etc. experimental study of using icy yellow detumescence analgesic to treat rabbit acute soft tissue injury, journal of orthopedics and traumatology in china, 2008, 16 (8): 42-43.]The method is improved to carry out experiments for influencing the viscosity of the blood plasma. 20 big-ear white rabbits are randomly divided into 4 groups, namely nine gardenia red liniment groups, safflower oil groups, ethanol solution groups and normal saline control groups, wherein each group comprises 5 rabbits. Shaving the inner side of the upper part of the right rear thigh of each group of rabbits, and preparing skin, wherein the exposed area is about 6cm multiplied by 5 cm; fixing the thigh with wood plate, marking the striking action point at the thick part of skin muscle, and using striking rod (head area of striking rod is 2 cm)2Impact impulse is 2kg · s) is hit on the mark by free fall at a height of 60cm, causing acute soft tissue injury. The visible small bleeding points on the skin of the beating part are used as the success of molding.
1.4.2 methods of administration
After molding for 1h, the rabbit of each group is respectively and correspondingly coated with nine gardenia red liniment, safflower oil, 60% ethanol solution and normal saline solution at the striking part for treatment, and then uniformly coated and massaged for 5min by using a cotton swab, wherein the dosage is 0.2ml/kg each time and 2 times/day for 7 days.
1.4.3 plasma viscometry
On days 1, 2, 3, 5 and 7 after administration, peripheral blood of each rabbit was collected and subjected to plasma viscosity examination. Referring to the literature [ Wanzhou Wu, Liarmy, syringe sleeve siliconizing reagent and siliconizing method thereof, China patent, CN 200510111312.0, 2007-06-13 ], a siliconized 5ml syringe was used, 0.1ml of heparin sodium solution was accurately aspirated, shaken to uniformly disperse the heparin sodium solution in the syringe, 3ml of blood was slowly drawn from the auricular vein, injected into a 5ml centrifuge tube, and centrifuged in a centrifuge at 3000r/min for 15 min. Taking 2ml of supernatant, placing the supernatant in a full-automatic blood rheology rapid tester, and measuring the viscosity value of blood plasma.
1.4.4 local tissue IL-1 beta content determination
After the peripheral blood is taken at the 7 th day after the medicine is taken, all groups of rabbits are sacrificed, the skin of the struck part of the rabbits is cut by a scalpel, subcutaneous tissues are separated, muscle tissues with bluish purple color at the center of the struck part are cut, the muscle tissues are rinsed for 1min by using normal saline, water is absorbed by filter paper, 80mg of tissues are taken, the tissues are accurately weighed by an electronic analytical balance, the normal saline is quantitatively added according to the proportion of 20mg/ml, the mixture is slowly and uniformly ground for 15min until uniform serous fluid is obtained, the uniform serous fluid is transferred to a 5ml centrifugal tube, the centrifugal tube is placed in a centrifugal machine for centrifugation for 15min at 3000r/min, 2ml of supernatant fluid is taken, and the IL-1 beta content of the tissue homogenate fluid is measured strictly according to the operation steps of an IL-1 beta radioimmunoassay kit.
1.5 statistical treatment
The experimental results are mean ± standard deviation
Shows that statistical processing was performed using SPSS 21.0 software, differences between groups were compared using one-way analysis of variance,pairwise comparisons between groups by LSD-t test, with P<0.05 is statistically significant.
2 results
2.1 Effect on plasma viscosity test results
The plasma viscosity of the rabbit of the nine-ingredient gardenia red liniment group is respectively and obviously higher than that of the normal saline control group (P <0.05) on 3 days, 5 days and 7 days after treatment, and the plasma viscosity of the rabbit of the nine-ingredient gardenia red liniment group is respectively and obviously higher than that of the ethanol solution group (P <0.05) on 3 days and 5 days. However, at each time point after treatment, the plasma viscosity of the nine gardenia red liniment groups was lower than that of the normal safflower oil group, but the difference between the two groups was not statistically significant (P > 0.05). On day 7 after treatment, the plasma viscosity of safflower oil was significantly higher than that of the normal saline control group (P <0.05), respectively. The nine-ingredient gardenia red liniment is shown to be capable of remarkably reducing the plasma viscosity of the rabbits with acute soft tissue injuries, and can take effect by applying more days, but the application time of the effect taking is shorter than that of the safflower oil. The results are shown in Table 2.
TABLE 2 measurement of rabbit plasma viscosity
Note: compared with the normal saline control group,*P<0.05，**P<0.01; compared with the ethanol solution group, the method has the advantages that,#P<0.05
2.2 Experimental results for influencing IL-1 beta content in local tissues
On the 7 th day after treatment, the IL-1 beta content of the local tissues of the nine-ingredient gardenia red liniment group rabbits is respectively and obviously lower than that of a normal saline control group and an ethanol solution group (P is less than 0.05); the IL-1 beta content of the nine-ingredient gardenia red liniment group is lower than that of the normal safflower oil group, but the difference between the two groups has no statistical significance (P is more than 0.05). The content of IL-1 beta in the safflower oil group is obviously lower than that in a normal saline control group (P < 0.05). The results show that the nine-ingredient gardenia red liniment can obviously reduce the IL-1 beta content of local tissues of rabbits, and the action intensity of the liniment is equivalent to that of the normal safflower oil, and the results are shown in a table 3.
TABLE 3 measurement results of IL-1. beta. content in rabbit local tissue
Note: compared with the normal saline control group,**P<0.01; compared with the ethanol solution group, the method has the advantages that,##P<0.01
3 conclusion
The nine-ingredient gardenia red liniment can obviously reduce the plasma viscosity and the IL-1 beta content of local tissues of rabbits with acute soft tissue injuries. After acute soft tissue injury occurs, the nine-ingredient gardenia red liniment is continuously applied externally for treatment at an early stage, so that the rheological property and blood circulation of blood can be improved, and swelling and blood stasis of an affected part can be effectively relieved; obviously inhibit the generation of local tissue inflammatory factors, inhibit early inflammatory exudation, relieve local pain symptoms and provide a good tissue repair environment for the injured part.
Comprehensively considered, the action mechanism of the nine-ingredient gardenia red liniment for treating acute soft tissue injury is possibly related to reducing the high blood viscosity state, inhibiting platelet aggregation, preventing thrombosis, expanding local capillary vessels, accelerating blood flow speed, improving microcirculation, inhibiting secretion of inflammatory factors, promoting absorption of inflammation and dilution operation of inflammatory mediators, inhibiting local pain and the like.
Example 4: clinical curative effect of treating acute soft tissue injury by applying nine-ingredient gardenia red liniment
1 data and method
1.1 case selection and grouping
Selecting 98 college patients with acute soft tissue injury on legs after movement of a certain college who visits the clinic of the college as study objects in total, wherein 72 male students and 26 female students are selected; the age is 18-26 years, and the average age (20.91 +/-1.70) years. Before the study began, the subjects of the two groups of patients were informed of the purpose, method and content of the study, and informed consent was obtained. By adopting a test control method, 98 patients are randomly divided into nine gardenia red liniment groups and positive safflower oil groups, wherein 48 patients in the nine gardenia red liniment groups and 50 patients in the positive safflower oil groups. Compared with the prior art, the sex distribution, the age and the difference of the injured parts of the two groups of patients have no statistical significance (P >0.05), and have consistency.
1.2 case inclusion criteria and exclusion criteria
1.2.1 case inclusion criteria
All patients had a clear history of trauma in their legs; the damaged part meets the diagnosis standard of acute soft tissue injury; swelling and pain or ecchymosis at the injury part, and accompanied with movement disorder; the course of disease does not exceed 1 day, and the patients are not treated by oral administration, external application of treatment medicines and other methods; the age is between 16 and 70 years old; no nerve and blood vessel damage; diseases of the heart, liver, kidney and hematopoietic system, mental diseases, tuberculosis, tumor, etc.
1.2.2 case exclusion criteria
Those under 15 years of age or above 71 years of age; pregnant or lactating women, people with allergic constitution; after X-ray radiography examination, orthopedists diagnose the patients with fracture and dislocation; wounds exist in the injured part, or other organ injuries are combined; completely broken soft tissues such as muscles, tendons, and ligaments; skin injury, skin eczema or pain, red swelling, and bluish purple due to inflammatory infection; if the drug is not administered according to the prescription, the curative effect or the data are not complete and the effect or the safety is not judged. Any one of the above is excluded.
1.3 methods of treatment
1.3.1 test drugs
Jiuweizhihong liniment (Jiuweizhihong liniment) produced by people hospital pharmacy in Taixing city with specification of 100 ml/bottle and batch number 181202. Regular safflower oil (red flower oil), manufactured by Wanyuan (Fuzhou) pharmaceutical Co., Ltd, having a specification of 20 ml/bottle, lot number 110906.
1.3.2 methods of use
The medicine is used for the first time within 24-48 h after injury, the medicine is smeared on the skin of soft tissue injury, nine gardenia red liniment are smeared on patients with nine gardenia red liniment groups, and normal safflower oil is smeared on patients with normal safflower oil for 2 times/day, wherein the dosage is 0.5ml/cm per time2Five days is a course of treatment. Observing and recording the symptom change of the injury part of two groups of patients on days 1, 3 and 5 before and after treatment; after one course of treatment, the clinical efficacy of the medicine is judged.
1.4 clinical symptom Observation index
1.4.1 extravasated blood
Subcutaneous extravasated blood is more than 2cm multiplied by 2cm and is counted as level II; subcutaneous extravasated blood is less than 2cm multiplied by 2cm and is marked as I grade; the color is normal and is rated as 0.
1.4.2 swelling
Muscle swelling is obvious, and deep fingerprints are marked after the finger pressure is 30s and are counted as level II; moderate swelling, obvious fingerprint after pressing with fingers for 30s, and is classified as grade I; there was no swelling, no fingerprint after 30s of finger pressure, and was rated 0.
1.4.3 pain
Severe pain, intolerable pain, no pain and no pain, and is counted as grade IV; the pain is very painful, the pain is more severe and the pain is aggravated by light touch, and the pain is rated as grade III; mild pain, limited pain, no pain, rated as level II; slight pain, similar to pain and non-pain, often appears in combination with other feelings, such as itching, tingling, heaviness, discomfort and the like, and is rated as I; no pain: no pain, no pain on touch, and a rating of 0.
1.5 determination of clinical efficacy
The clinical curative effect is divided into effective, effective and ineffective according to the improvement degree of the injury symptom. Wherein, the pain and the swelling are obviously relieved or disappeared, the blood circulation disorder is obviously improved, the extravasated blood is not increased and begins to be absorbed; the obvious effects are that the pain, the swelling, the blood circulation disorder and the extravasated blood are improved to a certain extent; the ineffectiveness was no significant change in pain, swelling, impaired blood circulation and congestion. The total effective rate of the treatment is calculated by the sum of effective and significant number of people.
1.6 statistical methods
Experimental results the data were measured as mean. + -. standard deviation
Representing, the counting data is represented by category grouping counting, the grade data is represented by grade grouping counting, SPSS 21.0 software is used for statistical processing, the nonparametric test is adopted to compare the difference between the symptoms of congestion, swelling and pain of the nine-ingredient gardenia red liniment group and the positive safflower oil group, and Chi is adopted2The clinical curative effect of the nine-ingredient gardenia red liniment group and the positive safflower oil group is compared by testing, and P is used<0.05 is statistically significant.
2 results
2.1 clinical symptom Change before and after treatment of two groups
The difference of the symptoms of congestion, swelling and pain of the patients in the two groups before treatment has no statistical significance (P is more than 0.05). Compared with the prior treatment, the symptoms of congestion, swelling and pain of the two groups of patients on the 1 st day after the treatment are not obviously improved. After 1 day of treatment, the pain symptom of the nine-ingredient gardenia red liniment is better than that of the normal safflower oil (P is less than 0.05). On the 3 rd day after treatment, the symptoms of congestion, swelling and pain of the two groups of patients are improved to a certain extent; the swelling and pain symptoms of the nine-ingredient gardenia red liniment group are better than those of the safflower oil (P is less than 0.05). On the 5 th day after treatment, the symptoms of congestion, swelling and pain of the two groups of patients continue to improve; the swelling symptom of the nine-ingredient gardenia red liniment group is better than that of the safflower oil (P <0.05), and the pain symptom is obviously better than that of the safflower oil (P < 0.01). Compared with regular safflower oil, the nine-ingredient gardenia red liniment has better effect on treating swelling and pain symptoms caused by acute soft tissue injury, and the results are shown in tables 4, 5 and 6.
TABLE 4 change in extravasated blood symptoms before and after treatment in two groups of patients
TABLE 5 Change in swelling symptoms before and after treatment in two groups of patients
TABLE 6 changes in pain symptoms before and after treatment in two groups of patients
2.2 comparison of clinical efficacy
After a treatment course, the total treatment effective rate of the patients with the nine-ingredient gardenia red liniment group is higher than that of the patients with the normal safflower oil group, and the difference has statistical significance (P is less than 0.05). The clinical efficacy of the nine-ingredient gardenia red liniment for treating acute soft tissue injury is better than that of the regular safflower oil, and the results are shown in table 7.
TABLE 7 clinical efficacy of two groups of patients
3 conclusion
The nine-ingredient gardenia red liniment has better effects of detumescence and acesodyne, and the clinical curative effect of treating acute soft tissue injury is better than the common safflower oil. The nine-ingredient gardenia red liniment is an ideal external medicine for treating acute soft tissue injury.
Claims (5)
1. The nine-ingredient gardenia red liniment for treating acute soft tissue injury is characterized by being prepared from the following components in parts by mass: 700-1300 parts of cape jasmine, 700-1300 parts of safflower, 350-650 parts of clematis root, 350-650 parts of angelica, 210-390 parts of ground beetle, 210-390 parts of borneol, 70-130 parts of frankincense, 70-130 parts of myrrh and 70-130 parts of dragon's blood.
2. The nine-ingredient gardenia red liniment for treating acute soft tissue injury according to claim 1, which is prepared from the following ingredients in parts by mass: 800-1200 parts of cape jasmine, 800-1200 parts of safflower, 400-600 parts of radix clematidis, 400-600 parts of angelica sinensis, 240-360 parts of ground beetle, 240-360 parts of borneol, 80-120 parts of frankincense, 80-120 parts of myrrh and 80-120 parts of dragon's blood.
3. The nine-ingredient gardenia red liniment for treating acute soft tissue injury according to claim 1, which is prepared from the following ingredients in parts by mass: 900-1100 parts of cape jasmine, 900-1100 parts of safflower, 450-550 parts of clematis root, 450-550 parts of angelica, 270-330 parts of ground beetle, 270-330 parts of borneol, 90-110 parts of frankincense, 90-110 parts of myrrh and 90-110 parts of dragon's blood.
4. The preparation method of the nine-ingredient gardenia red liniment for treating acute soft tissue injury according to any one of claims 1 to 3, which is characterized in that the nine-ingredient gardenia red liniment is prepared according to the following preparation method: weighing the medicinal materials of gardenia, safflower, clematis root, angelica, ground beeltle, frankincense, myrrh and dragon blood according to the prescription proportion, uniformly mixing, crushing and sieving; soaking 1g of medicinal materials in 70-90% ethanol with the volume of 4-6 times of that of 1g of medicinal materials to 1ml of solvent for 48 hours, slowly percolating, collecting initial percolate for later use, adding 70-90% ethanol with the volume of 4-6 times of that of the medicinal materials to slowly percolate, concentrating the percolate under reduced pressure until no alcohol smell exists, mixing the percolate with the initial percolate, adding borneol with the crushed formula amount proportion, adding a proper amount of 95% ethanol to uniformly disperse, adding pure water to adjust the total amount of the solution to be 6.21-6.61 times of the total amount of the medicinal materials by volume of ethanol solution, shaking uniformly, and subpackaging to obtain the traditional Chinese medicine.
5. The method for preparing the nine-ingredient gardenia red liniment for treating acute soft tissue injury according to claim 4, wherein gardenia, safflower, clematis root, angelica, ground beetle, frankincense, myrrh and dragon's blood are mixed evenly, crushed and sieved by a 80-mesh sieve; according to the ratio of 1g of medicinal materials to 1ml of solvent, firstly soaking the medicinal materials in 80% ethanol with the volume 5 times that of the medicinal materials for 48 hours, slowly percolating the medicinal materials, collecting initial percolate for later use, then adding 80% ethanol with the volume 5 times that of the medicinal materials for slowly percolating, concentrating the percolate under reduced pressure until no alcohol smell exists, then combining the percolate with the initial percolate, adding borneol with the formula amount ratio after crushing, adding 95% ethanol with the volume 0.17 time that of the total amount of the medicinal materials for uniform dispersion, then adding pure water to adjust the total amount of the solution to be 6.41 times of the total amount of the medicinal materials in volume of ethanol solution, shaking the mixture uniformly, adjusting the density to be 0.90-0.94 g/ml, and subpackaging the medicinal materials to obtain the composition.
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