A kind of anti-inflammatory and analgesic Li medicine and preparation method

Abstract

On the basis of a traditional Li medicine for relieving cervical spondylosis, a new Li medicine for relieving cervical spondylosis was developed by adding copper pen flower fruit extract. The copper pen flower fruit extract has bergenin, various phenolic acids and alcohol compounds as the main active ingredients, and has analgesic and anti-inflammatory activities. By improving the traditional Li medicine prescription, a new Li medicine for relieving cervical spondylosis was developed. After the hot plate pain threshold reaction experiment on mice, the acetic acid-induced painful writhing experiment on mice, and the anti-inflammatory experiment on xylene-induced ear swelling in mice, it is fully proved that the copper pen flower fruit extract, the traditional Li medicine for relieving cervical spondylosis, and the new Li medicine compounded with the copper pen flower fruit extract and traditional Li medicine all have the effects of dredging meridians, promoting blood circulation and removing blood stasis, reducing swelling and relieving pain, and dispelling wind and cold, and have anti-inflammatory and analgesic effects. The analgesic and anti-inflammatory effects of the new Li medicine prescription for relieving cervical spondylosis are better than those of the copper pen flower fruit extract and the traditional Li medicine prescription for relieving cervical spondylosis.

Description

Technical Field

[0001] The invention relates to a novel Li medicine for relieving cervical spondylosis and a preparation method thereof, and belongs to the field of ethnic medicine. Background Art

[0002] A traditional Li medicine prescription for relieving cervical spondylosis is a prescription of Wu Qianhua, a Li doctor in Tunchang County, Hainan Province. The prescription consists of 10 Li medicinal materials, including Millettia reticulata, Rhizoma Atractylodis Macrocephalae, Eucommia ulmoides, Caulis lappa, Caulis dinggong, Bidens pilosa, Rhizoma lycopodii, Herba striatae, Folium banyan, and Agarwood. Its functions and indications are to relax muscles and activate collaterals, reduce swelling and relieve pain. It is mainly used to treat pain in the cervical spine, shoulders, waist and legs, and injuries from falls.

[0003] Li medicine has a history of more than 3,000 years on Hainan Island. It has solved the problems faced by the Li people due to inconvenient transportation, lack of information, and lack of medical care and medicine. It is a summary of the experience accumulated by the Li people in their struggle with nature in life. It is a precious heritage of the Li medicine culture. According to the symptoms, there are two treatment methods: internal and external application. It is effective for rheumatism, pain in the neck, shoulders, waist and legs, postpartum congestion, traumatic injuries and other symptoms.

[0004] The Li people only have language but no writing. The inheritance of Li medicine used to rely on word of mouth from generation to generation, and there is no systematic Li medicine theory system with written expression. As time goes by, there are fewer and fewer experienced old Li doctors, and the inheritance is also facing a crisis. Therefore, the traditional Li medicine prescriptions need rescue protection and resource excavation, and prescription compilation is imminent. When Li medicine is used externally, most of them adopt the traditional preparation method, and apply the affected area with the fresh leaves of Li medicine after heating or stir-fry the whole plant of Li medicine in a pot and apply it to the affected area. The disadvantage is that the effective ingredients of the medicine are easy to lose, easy to mold, and difficult to preserve. Slice the roots of Li medicine, dry and crush the leaves, stems, fruits, and flowers, mix with a little honey and rice wine, put them into a cloth bag and steam for 20-30 minutes, and apply them to the affected area when the skin is not scalded to achieve the purpose of treatment. This method is cumbersome to operate, lacks quality standards, has poor controllability, and is not easy to transport outside the island and sell to the international market, which limits its use and dissemination. Based on further pharmacological and efficacy experimental analysis, the inventor believes that the original formula still has shortcomings and needs to be further improved.

[0005] Ardisia obtusa Mez (with Figure 1 ) is an evergreen shrub of the genus Ardisia of the Myrsinaceae family. Literature reports that Ardisia plants have the effects of clearing away heat and dampness, relieving swelling and pain, and promoting blood circulation and removing blood stasis, and have obvious effects on traumatic injuries, rheumatism, tuberculosis and various inflammations. Copperpen flower also has a long history of application in the Li ethnic group, mainly used for the treatment of traumatic injuries, rheumatism, sore throat and other diseases. Studies have found that its fruit extract has the effect of anti-inflammatory and analgesic. Therefore, the present invention compounded the above-mentioned traditional Li medicine with the copperpen flower fruit extract to develop a new Li medicine for relieving cervical spondylosis.

[0006] After literature review, there is no report on the use of copper pen flower fruit extract in the above-mentioned medical aspects. Summary of the invention

[0007] The purpose of the present invention is to provide a novel herbal medicine for relieving cervical spondylosis and a preparation method thereof. In order to achieve the above purpose, the present invention adopts the following technical scheme.

[0008] The invention discloses a novel Li medicine for relieving cervical spondylosis and a preparation method thereof, comprising the following steps:

[0009] 1. Preparation of the Herba Ligustici Ervatamiae Extract

[0010] Millettia spatholobi extract: Take 1kg of dried Millettia spatholobi stems and crush them with a grinder. Place the crushed medicinal material in a reflux extraction tank. Add 50% (V / V) ethanol and reflux for 3 times at 60°C, reflux for 1 hour each time. Add 3L of the above ethanol for the first time, 2L for the second time, and 1L for the third time. Combine the three extracts, distill and recover ethanol under reduced pressure, concentrate the extract, and finally freeze-dry the extract to obtain the extract.

[0011] Extract of Drosophila serrata: Dry the collected Drosophila serrata stems in the shade, grind them with a pulverizer, and store the obtained plant powder in a sealed bag in a light-shielded place for later use. Weigh 1 kg of the sample, place it in a reflux extraction tank, add 50% (V / V) ethanol and reflux for 3 times at 60°C, reflux for 1 hour each time. Add 3L of the above ethanol for the first time, 2L for the second time, and 1L for the third time. Combine the three extracts, distill and recover the ethanol under reduced pressure, concentrate the extract, and finally freeze-dry the extract to obtain the extract.

[0012] Eucommia ulmoides extract: Weigh the dried Eucommia ulmoides stem bark and crush it. 1 kg of the obtained powder is placed in a reflux extraction tank. At 60°C, add 50% (V / V) ethanol and reflux for extraction for 3 times, each time for 1 hour. Add 3L of the above ethanol for the first time, 2L for the second time, and 1L for the third time. Combine the three extracts, distill and recover the ethanol under reduced pressure, concentrate the extract, and finally freeze-dry the extract to obtain the extract.

[0013] Extract of Caulis Sinensis: Take 1kg of dried Caulis Sinensis stems, crush them, and place them in a reflux extraction tank. Add 50% (V / V) ethanol and reflux for 3 times at 60°C, reflux for 1h each time. Add 3L of the above ethanol for the first time, 2L for the second time, and 1L for the third time. Combine the three extracts, distill and recover ethanol under reduced pressure, concentrate the extract, and finally freeze-dry the extract to obtain the extract.

[0014] Dinggongteng extract: Take 1kg of dried Dinggongteng stems, crush them, put them in a reflux extraction tank, add 50% (V / V) ethanol and reflux for 3 times at 60°C, reflux for 1h each time. Add 3L of the above ethanol for the first time, 2L for the second time, and 1L for the third time. Combine the three extracts, distill and recover ethanol under reduced pressure, concentrate the extract, and finally freeze-dry the extract to obtain the extract.

[0015] Bidens pilosa extract: Weigh 1kg of dry whole herb powder of Bidens pilosa, place it in a reflux extraction tank, add 50% (V / V) ethanol and reflux for 3 times at 60°C, reflux for 1h each time. Add 3L of the above ethanol for the first time, 2L for the second time, and 1L for the third time. Combine the three extracts, distill and recover ethanol under reduced pressure, concentrate the extract, and finally freeze-dry the extract to obtain the extract.

[0016] Extract of Rhizoma Dioscoreae: Take 1kg of dried Rhizoma Dioscoreae root, crush it, put it in a reflux extraction tank, add 50% (V / V) ethanol and reflux for 3 times at 60℃, reflux for 1h each time. Add 3L of the above ethanol for the first time, 2L for the second time, and 1L for the third time. Combine the three extracts, distill and recover ethanol under reduced pressure, concentrate the extract, and finally freeze-dry the extract to obtain the extract.

[0017] Extract of Herba Lycopodii: Take the dried Herba Lycopodii whole plant and grind it. 1 kg of the powder after grinding is placed in a reflux extraction tank. At 60°C, add 50% (V / V) ethanol for reflux extraction for 3 times, and reflux for 1 hour each time. Add 3L of the above ethanol for the first time, 2L for the second time, and 1L for the third time. Combine the three extracts, distill and recover ethanol under reduced pressure, concentrate the extract, and finally freeze-dry the extract to obtain the extract.

[0018] Banyan tree whisker extract: Take dried banyan tree whisker branches and crush them. After crushing, weigh 1 kg of powder, put it in a reflux extraction tank, add 50% (V / V) ethanol and reflux for 3 times at 60°C, reflux for 1 hour each time. Add 3L of the above ethanol for the first time, 2L for the second time, and 1L for the third time. Combine the three extracts, distill and recover ethanol under reduced pressure, concentrate the extract, and finally freeze-dry the extract to obtain the extract.

[0019] Agarwood extract: Select agarwood that has formed fragrance, wash it, dry it, and crush it into 20-40 meshes. Put the agarwood powder into a supercritical carbon dioxide extraction device for extraction to obtain an extract, wherein the anhydrous ethanol used in the extraction is used as an entrainer, and the amount of anhydrous ethanol added is 15% of the weight of the agarwood. In the step, the supercritical carbon dioxide extraction temperature is 35-50°C, the extraction pressure is 20-40MPa, the carbon dioxide flow rate in the supercritical carbon dioxide extraction is 25L / h, the extraction time is 4-6h, and the extraction device is released after removing impurities in the extract, and the agarwood essential oil and the extract are separated. The extract is filtered and purified to obtain the agarwood extract.

[0020] Copperpen Flower Fruit Extract: Copperpen Flower was collected from the back mountain of Lingzai Village, Fengmu Town, Tunchang County, Hainan Province. It was jointly identified as Copperpen Flower by Professor Zeng Niankai, a doctoral student at Hainan Normal University, and Researcher Wang Yong of the School of Pharmacy of Hainan Medical College. The dried Copperpen Flower fruit was crushed, 1 kg was weighed out, and placed in a reflux extraction tank. At 60°C, 50% (V / V) ethanol was added for reflux extraction for 3 times, each time for 1 hour. 3L of the above ethanol was added for the first time, 2L for the second time, and 1L for the third time. The three extracts were combined, ethanol was distilled and recovered under reduced pressure, the extract was concentrated, and finally the extract was freeze-dried to obtain the extract.

[0021] The extracts of the root, fruit and leaf of Copper Pen Flower were tested by the Analysis and Testing Center of the Institute of Deep Sea Science and Engineering, Chinese Academy of Sciences. The test methods and conditions are as follows:

[0022] (1) Sample preparation: Take 0.2 g of each extract, add 40 mL of 75% methanol to dissolve, centrifuge at 5000 r / min for 5 min, filter through a 0.22 μm filter membrane, and take 1 mL of the solution after filtering for detection.

[0023] (2) Chromatographic conditions: column temperature 30°C, flow rate set at 0.3 mL / min, injection volume 1 μL, mobile phase methanol (A)-0.1% formic acid aqueous solution (B), gradient elution: 0-2 min, 17% A; 2-3.5 min, 17%-23% A; 3.5-6 min, 23%-45% A; 6-13 min, 45%-90% A; 13-15 min, 90% A; 15-16 min, 90%-17% A; 16-19 min, 17% A

[0024] (3) Mass spectrometry conditions: positive and negative ion modes were used, voltage was 3.0 kV, ion transfer tube temperature was 320 °C, ion source temperature was 350 °C, scanning mode was Full MS / dd-MS 2 , Full MS resolution 70,000, dd-MS 2 The resolution was 17500, the scanning range was m / z 80-1200, and the collision energy (NCE) was 30%.

[0025] The test results are as attached Figure 2 As shown, the copper basin flower fruit is rich in bergenin (attached Figure 2 B) Anti-inflammatory and analgesic ingredients such as various acids and alcohols.

[0026] 2. Preparation of a new type of Li medicine for relieving cervical spondylosis

[0027] Implementation Plan 1

[0028] 45% of copper pot flower fruit extract, 7.5% of Millettia spatholobi extract, 5% of Croton tiglium extract, 5% of Eucommia ulmoides extract, 7.5% of Croton tiglium extract, 5% of Trichosanthes kirilowii extract, 5% of Bidens pilosa extract, 5% of Rhizoma Dioscoreae extract, 5% of Lycopodiella ciliata extract, 5% of Banyan tree extract, and 5% of Aquilaria woodii extract.

[0029] The total weight reaches 100%, and the above drugs are ground into powder, fully mixed and made into tablets, each tablet is 1g, and 2 tablets are taken orally before each meal, 3 times a day.

[0030] Implementation Plan 2

[0031] 45g of copper pot flower fruit extract, 7.5g of Millettia spatholobi extract, 5g of Croton tiglium extract, 5g of Eucommia ulmoides extract, 7.5g of Croton tiglium extract, 5g of Trichosanthes kirilowii extract, 5g of Bidens pilosa extract, 5g of Rhizoma Dioscoreae extract, 5g of Herba Lycopodii extract, 5g of Herba Lycopodii extract, 5g of Banyan tree whisker extract, and 5g of Aquilaria wood extract.

[0032] The total weight reaches 100g, and the above drugs are ground into powder, mixed thoroughly and made into tablets, each tablet is 1g, and the patient takes 2 tablets orally before each meal, 3 times a day.

[0033] Implementation Plan 3

[0034] 45g of copper pot flower fruit extract, 7.5g of Millettia spatholobi extract, 5g of Croton tiglium extract, 5g of Eucommia ulmoides extract, 7.5g of Croton tiglium extract, 5g of Trichosanthes kirilowii extract, 5g of Bidens pilosa extract, 5g of Rhizoma Dioscoreae extract, 5g of Herba Lycopodii extract, 5g of Herba Lycopodii extract, 5g of Banyan tree whisker extract, and 5g of Aquilaria wood extract.

[0035] The total weight reaches 100g, and the above drugs are ground into powder, mixed thoroughly and evenly to make a powder. Take an appropriate amount of the powder, dissolve it in water and apply it to the affected area 3 times a day.

[0036] Implementation Plan 4

[0037] 45g of copper pot flower fruit extract, 7.5g of Millettia spatholobi extract, 5g of Croton tiglium extract, 5g of Eucommia ulmoides extract, 7.5g of Croton tiglium extract, 5g of Trichosanthes kirilowii extract, 5g of Bidens pilosa extract, 5g of Rhizoma Dioscoreae extract, 5g of Herba Lycopodii extract, 5g of Herba Lycopodii extract, 5g of Banyan tree whisker extract, and 5g of Aquilaria wood extract.

[0038] The total weight reaches 100g, and the above drugs are ground and mixed evenly to make powder, and then 100g of ordinary honey is added to fully blend, and evenly divided into 50 portions to make honey pills, each weighing 4g. The patient takes 1 pill orally before each meal, 3 times a day.

[0039] Implementation Plan 5

[0040] 45g of copper pot flower fruit extract, 7.5g of Millettia spatholobi extract, 5g of Croton tiglium extract, 5g of Eucommia ulmoides extract, 7.5g of Croton tiglium extract, 5g of Trichosanthes kirilowii extract, 5g of Bidens pilosa extract, 5g of Rhizoma Dioscoreae extract, 5g of Herba Lycopodii extract, 5g of Herba Lycopodii extract, 5g of Banyan tree whisker extract, and 5g of Aquilaria wood extract.

[0041] The total weight reaches 100g, and the above-mentioned drugs are ground into powder, mixed thoroughly and evenly, made into powder, and loaded into medicinal glutinous rice capsules. The net weight of each capsule is 0.5g. The patient takes 4 capsules orally before each meal, 3 times a day.

[0042] Implementation Plan 6

[0043] 45g of copper pot flower fruit extract, 7.5g of Millettia spatholobi extract, 5g of Croton tiglium extract, 5g of Eucommia ulmoides extract, 7.5g of Croton tiglium extract, 5g of Trichosanthes kirilowii extract, 5g of Bidens pilosa extract, 5g of Rhizoma Dioscoreae extract, 5g of Herba Lycopodii extract, 5g of Herba Lycopodii extract, 5g of Banyan tree whisker extract, and 5g of Aquilaria wood extract.

[0044] The total weight reaches 100g, and the above medicines are ground into powder, mixed thoroughly and evenly, made into granules, packed in small medicinal paper bags and sealed. The net weight of each bag is 2g. The patient takes 1 bag before each meal, 3 times a day.

[0045] Implementation Plan 7

[0046] According to the following weight ratio: 45% of copper pot flower fruit extract, 7.5% of Millettia spatholobi extract, 5% of Croton tiglium extract, 5% of Eucommia ulmoides extract, 7.5% of Croton tiglium extract, 5% of Trichosanthes kirilowii extract, 5% of Bidens pilosa L. extract, 5% of Rhizoma Dioscoreae extract, 5% of Lycopodiella cuneata extract, 5% of Banyan tree extract, and 5% of Aquilaria woodii extract.

[0047] The above-mentioned drugs are ground into fine powder, mixed thoroughly and evenly, and made into external preparations such as sprays, tinctures, creams, gels, ointments, plasters, etc.

[0048] Attached Figure 1 :Copperpen flower (A) and copperpen flower fruit (B) copperpen flower root (C) copperpen flower leaf (D)

[0049] Attached Figure 2: LC-MS chromatograms of extracts from the roots, fruits and leaves of Copper Pen Flower (A is the root, B is the fruit, which is rich in bergenin, and C is the leaf).

[0050] Attached Figure 3 : Mouse hot plate pain threshold reaction experiment: Group A was a blank control group (normal saline), Group B was an indomethacin control group, Group C was an experimental group of copper pen flower fruit extract, Group D was a control group of traditional Li medicine for relieving cervical spondylosis; Group E was an experimental group of a new Li medicine for relieving cervical spondylosis.

[0051] Attached Figure 4 :Anti-inflammatory experiment on ear swelling induced by xylene in mice: (Attachment Figure 4 A) The right ears of mice in group A were smeared with xylene on the front and back and the ears were obviously swollen by naked eye observation; (Attachment Figure 4 After xylene was applied to the front and back of the right ear of mice in group B, the ears of mice were slightly swollen by naked eye observation; (Attachment Figure 4 C) The right ears of mice in group C were smeared with xylene on both sides and the swelling was mild after visual observation; (Attachment Figure 4 The degree of swelling of mice in group D, which was observed by naked eye after applying xylene on the front and back of the right ear, was less than that in group C; (Attachment Figure 4 The swelling degree of mice in group E was less severe and close to that in group B when xylene was applied on the front and back of the right ears and observed with naked eyes. DETAILED DESCRIPTION

[0052] 1. Experimental instruments and equipment

[0053] Electronic balance, Changzhou Wande Balance Instrument Co., Ltd. FA12D4G; YLS-6B intelligent hot plate instrument, Anhui Zhenghua Biological Instrument Equipment Co., Ltd.; supercritical extraction equipment HA221-50-06, Nantong Hua'an Supercritical Extraction Equipment Co., Ltd.; Junde brand freeze vacuum dryer FD-503; water bath, oven, incubator, refrigerator, rotary evaporator, centrifuge, clean bench.

[0054] 2. Materials and reagents

[0055] Materials: No. 12 gavage needle, syringe, round-bottom flask, Erlenmeyer flask, heat reflux device, funnel, gauze, measuring cylinder, petri dish, cotton swab, volumetric flask, pipette, suction bulb, 8mm hole puncher, mouse cage, feed, and sawdust for mice.

[0056] Reagents and drugs: glacial acetic acid, Guangdong Guanghua Science and Technology Co., Ltd. batch number 20200312; xylene, Xilong Science Co., Ltd., batch number 2310242; ethanol, normal saline; indomethacin, Yunpeng Pharmaceutical Group Co., Ltd., product batch number G220204; copper pen flower fruit extract, traditional Li medicine for relieving cervical spondylosis and new Li medicine for relieving cervical spondylosis are homemade in the laboratory.

[0057] Experimental animals: White mice, male and female, four weeks old, purchased from Changsha Tianqin Biotechnology Co., Ltd., batch number: 43072623010048425

[0058] 1. Experimental methods

[0059] (1) Drug preparation

[0060] Indomethacin: Grind 5 tablets of indomethacin (5×25 mg) and place in a 100 mL volumetric flask. Add purified water to fully dissolve and make up to the mark. Shake well and gavage 0.2 mL (containing 0.25 mg of drug) to each mouse.

[0061] Copper Pen Flower Fruit Extract: Accurately weigh 0.6g and place it in a 100mL volumetric flask, add purified water to the mark, fully dissolve and shake well, and administer 0.2mL (containing 1.2mg of the drug) to each mouse by gavage.

[0062] Traditional Li medicine for relieving cervical spondylosis: Accurately weigh 0.6g of traditional Li medicine and place it in a 100mL volumetric flask, add purified water to the mark, fully dissolve and shake well, and gavage 0.2mL (containing 1.2mg) to each mouse.

[0063] A new type of Li medicine for relieving cervical spondylosis: Accurately weigh 0.6 g of the new Li medicine and place it in a 100 mL volumetric flask, add purified water to the mark, fully dissolve and shake well, and gavage 0.2 mL (containing 1.2 mg) to each mouse.

[0064] (2) Animal grouping and drug administration

[0065] 70 healthy female mice weighing between 18-22 g were selected (those that licked their hind feet within 5 seconds or did not lick their hind feet within 30 seconds were discarded). 50 mice were retained after screening and divided into 5 groups, with 10 mice in each group.

[0066] A: The blank control group (normal saline) was intragastrically administered with 10 mL / kg body weight of normal saline, 0.2 mL per mouse.

[0067] B: Indomethacin control group 0.2mL (containing 0.25mg of drug).

[0068] C: Experimental group of copper pen flower fruit extract, each mouse was gavaged with 0.2 mL (containing 1.2 mg of drug).

[0069] D: The control group of traditional Li medicine for relieving cervical spondylosis, each mouse was gavaged with 0.2 mL (containing 1.2 mg of medicine).

[0070] E: Experimental group of the new Li medicine for relieving cervical spondylosis, each mouse was gavaged with 0.2 mL (containing 1.2 mg of the medicine).

[0071] Groups A, C, D, and E were gavaged once a day. For 6 consecutive days, group B indomethacin was gavaged only once with 0.2 mL (containing 0.25 mg of the drug).

[0072] (3) Hot plate method for thermal pain in mice

[0073] Adjust the intelligent hot plate instrument so that its temperature is controlled at 55°C and preheat for 30 minutes.

[0074] The pain threshold of mice in the blank control group (normal saline) was measured. One mouse was placed on a hot plate for observation each time. The reaction of the mouse licking its hind paw was recorded with a stopwatch. The average of 10 times was taken as the pain threshold index (see Appendix). Figure 3 A).

[0075] The pain threshold of mice in the indomethacin control group was measured. One mouse was placed on a hot plate for observation and recording using a stopwatch. The reaction of the mouse licking its hind paw was used as the pain threshold index, and the average value of 10 times was taken (see Appendix). Figure 3 B).

[0076] The pain threshold of mice in the C. tongpenhua fruit extract experimental group was measured. One mouse was placed on a hot plate for observation each time. The reaction of the mouse licking its hind paw was recorded with a stopwatch as the pain threshold index. The average value of 10 times was taken (see Appendix). Figure 3 C).

[0077] The pain threshold of mice in the control group of traditional Chinese medicine for relieving cervical spondylosis was measured. One mouse was placed on a hot plate for observation and a stopwatch was used to record the reaction of the mouse licking its hind paw as the pain threshold index, and the average value of 10 times was taken (see Appendix). Figure 3 D).

[0078] The pain threshold of mice in the experimental group of the new drug for relieving cervical spondylosis was measured. One mouse was placed on a hot plate for observation each time. The reaction of the mouse licking its hind paw was recorded with a stopwatch. The average of 10 times was taken (see Appendix). Figure 3 E).

[0079] The pain threshold indices of 30min, 60min, 90min, 120min, 150min and 180min were recorded in the five groups respectively, the differences of pain threshold in each time period were compared among the groups, and the analgesia rate was calculated.

[0080] Pain threshold increase percentage (%) = (pain threshold after administration - pain threshold before administration / pain threshold before administration) × 100%. The results were processed by t-test to calculate the significance of the difference between the administration group and the blank control group (normal saline).

[0081] The experimental results are attached. Figure 3 and Table 1.

[0082] Table 1 Results of the hot plate pain threshold response experiment in mice

[0083]

[0084] Note: Compared with the blank control group, *P<0.05, **P<0.01

[0085] As can be seen from Table 1, compared with the blank control group A (normal saline), the copper pot flower fruit extract group C, the traditional Li medicine group D for relieving cervical spondylosis, and the new Li medicine group E for relieving cervical spondylosis all had analgesic effects at 30min, 60min, 90min, and 120min (P < 0.05). Only the pain threshold increase rate of group D was low at 30min. From the perspective of pain threshold, the analgesic effect of the new Li medicine group E for relieving cervical spondylosis was equivalent to that of the indomethacin control group B. Experimental verification shows that the new Li medicine E for relieving cervical spondylosis has a significant analgesic effect.

[0086] (4) Acetic acid-induced writhing experiment in mice

[0087] 50 male mice were screened and divided into 5 groups, 10 mice in each group, with a body weight between 18-20g. A was a blank control group (normal saline), which was gavaged with 10 ml / kg body weight of normal saline; B was an indomethacin control group, in which each mouse was gavaged with 0.2 ml (containing 0.25 mg of the drug); C was an experimental group of copper-pen flower fruit extract, in which each mouse was gavaged with 0.2 ml (containing 1.2 mg of the drug); D was a control group of traditional Li medicine for relieving cervical spondylosis, in which each mouse was gavaged with 0.2 ml (containing 1.2 mg of the drug); and Group E was an experimental group of new Li medicine for relieving cervical spondylosis, in which each mouse was gavaged with 0.2 ml (containing 1.2 mg of the drug).

[0088] Groups A, C, D, and E were gavaged every day for 6 consecutive days. On the 6th day of the experiment, the control group B indomethacin was gavaged only once. 0.6% acetic acid solution was injected intraperitoneally 1 hour after the last administration of the drug, 0.2 ml / animal. The number of writhing reactions within 20 minutes after the injection of acetic acid solution was observed and recorded. After the experiment was completed, the percentage of drug analgesia was calculated. The writhing reaction was manifested as a sunken abdomen, swollen trunk and hind limbs, and raised buttocks.

[0089] Analgesia rate = (average number of writhing times in the control group - average number of writhing times in the drug-treated group) / average number of writhing times in the control group × 100%.

[0090] Table 2 Results of the acetic acid-induced writhing experiment in mice

[0091]

[0092] Note: Compared with the blank control group, *P<0.05, **P<0.01

[0093] Analysis of results: Compared with the blank control group A (normal saline), the indomethacin control group B, the copper-pen flower fruit extract experimental group C, the traditional Li medicine control group D for relieving cervical spondylosis, and the new Li medicine experimental group E for relieving cervical spondylosis, the number of body twisting of mice was significantly reduced (P < 0.05), indicating that the indomethacin control group, the copper-pen flower fruit extract, the traditional Li medicine for relieving cervical spondylosis, and the new Li medicine for relieving cervical spondylosis have an inhibitory effect on the abdominal pain of mice caused by acetic acid, among which the inhibitory effect of the new Li medicine experimental group for relieving cervical spondylosis on the abdominal pain of mice caused by acetic acid is slightly lower than that of the indomethacin group B, but higher than that of the C and D groups.

[0094] (5) Anti-inflammatory experiment on xylene-induced ear swelling in mice

[0095] First, 50 male mice weighing between 18-22 g were taken and divided into 5 groups, with 10 mice in each group.

[0096] The specific experimental methods are as follows:

[0097] Group A: Each mouse was intragastrically administered with 0.2 mL of normal saline (10 mL / kg body weight) for 6 consecutive days.

[0098] Group B: Indomethacin control group. On the sixth day of the experiment, each mouse was gavaged with 0.2 mL (containing 0.25 mg of the drug) once per day.

[0099] Group C: The fruit extract of Copper Pen Flower was administered intragastrically 0.2 mL (containing 1.2 mg) per mouse every day for 6 consecutive days.

[0100] Group D: Traditional Li medicine for relieving cervical spondylosis, 0.2 mL (containing 1.2 mg) was gavaged into each mouse every day for 6 consecutive days.

[0101] Group E: A new type of Li medicine for relieving cervical spondylosis was administered intragastrically with 0.2 mL (containing 1.2 mg) per mouse per day for 6 consecutive days.

[0102] One hour after the last administration, 0.2 mL of xylene was applied to the front and back of the right ear of each mouse, and the left ear was used as a blank control. The degree of swelling of the right ear of each group of mice was observed with the naked eye. After 25 minutes, the mice were killed by cervical dislocation, and both ears were cut off along the baseline of the auricle. A circular ear piece was punched at the same position of both ears with an 8 mm diameter puncher. The weight of the left and right ear pieces was weighed, and the difference in weight between the left and right ear pieces was taken as the degree of swelling. The degree of swelling and inhibition rate were calculated.

[0103] The degree of ear swelling: the weight of the ear on the inflamed side (right ear) minus the weight of the ear on the control side (left ear).

[0104] Swelling inhibition rate = (auricle swelling rate of the control group - auricle swelling rate of the drug-treated group) / auricle swelling rate of the control group × 100%.

[0105] Results and analysis of the acute ear swelling experiment in mice (see Appendix Figure 4 The right ears of mice in group A were smeared with xylene and the swelling was obvious after naked eye observation (see attached Figure 4 A), after xylene was applied to the front and back of the right ear of mice in group B, the ears of mice were slightly swollen (see attached Figure 4 B, xylene was applied to the front and back of the right ear of mice in group C and the swelling was mild after visual observation (see Appendix Figure 4 C), xylene was applied to the front and back of the right ear of mice in group D and the degree of swelling was less than that in group C (Appendix Figure 4 D), xylene was applied to the front and back of the right ear of mice in group E and the swelling was observed with naked eyes. The degree of swelling was lighter and similar to that of group B (see Appendix Figure 4 E).

[0106] Table 3 Results of anti-inflammatory experiment on ear swelling in mice induced by xylene

[0107]

[0108] Note: Compared with the blank control group, *P<0.05, **P<0.01

[0109] The results showed that the copper pen flower fruit extract, traditional Li medicine for relieving cervical spondylosis, and new Li medicine for relieving cervical spondylosis all had anti-inflammatory effects, and the anti-inflammatory effect of the new Li medicine E for relieving cervical spondylosis was better than that of group C and group D, but lower than that of group B.

Claims

1. An anti-inflammatory and analgesic herbal medicine, characterized in that: The specific composition is as follows: 45% of copper pot flower fruit extract, 7.5% of Millettia spatholobi extract, 5% of Croton tiglium extract, 5% of Eucommia ulmoides extract, 7.5% of Croton tiglium extract, 5% of Trichosanthes kirilowii extract, 5% of Bidens pilosa extract, 5% of Rhizoma Dioscoreae extract, 5% of Eleutherodactylae extract, 5% of Banyan tree extract, and 5% of Aquilaria woodii extract; wherein, the extracts are all ethanol extracts.

2. The method for preparing Li medicine according to claim 1, characterized in that: The extracting method of the extract uses an ethanol solution, performs reflux leaching under heating conditions, then recovers the ethanol, and finally obtains the extract through freeze drying.

3. The Li medicine according to claim 1, or the Li medicine obtained by the preparation method according to claim 2, characterized in that: The above-mentioned Li medicine is made into one of gel, spray, patch, plaster or tincture.

Images