CN111557986B - Callicarpa nudiflora extract and application thereof in preparation of medicine for treating allergic purpura - Google Patents
Callicarpa nudiflora extract and application thereof in preparation of medicine for treating allergic purpura Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/85—Verbenaceae (Verbena family)
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/14—Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
Abstract
The invention provides a callicarpa nudiflora extract and application thereof in preparing a medicament for treating allergic purpura, belonging to the technical field of traditional Chinese medicine extraction. In the implementation process, the mixed solvent is adopted for extraction, the concentration of the extraction solvent is controlled, the volume ratio and the concentration of ethanol, acetone and petroleum ether are controlled, so that the extraction rate of the active ingredients of callicarpa nudiflora can be obviously improved, the total extraction rate of flavonoids can be obviously improved, the efficient release of the active ingredients can still be ensured at a lower temperature, the extraction efficiency is improved, and the waste of traditional Chinese medicines is reduced; the callicarpa nudiflora extract provided by the invention can be used for effectively treating anaphylactoid purpura, and is good in treatment effect, safe and effective.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine extraction, and in particular relates to a callicarpa nudiflora extract and application thereof in preparation of a medicine for treating allergic purpura.
Background
The Callicarpa nudiflora (root, stem, leaf of Callicarpa nudiflora (L.) Merr of Verbenaceae) contains condensed tannic acid, flavonoid glycoside, neutral resin, phenols, polysaccharides, hydroxyl compounds, magnesium, calcium, iron salt, etc., and has antiinflammatory, toxic substance removing, astringent, hemostatic, and pain relieving effects. Callicarpa nudiflora can reduce capillary permeability, has obvious function of contracting blood vessels, can inhibit the growth of staphylococcus aureus, streptococcus, salmonella typhi, pseudomonas aeruginosa and other bacteria, and can prevent secondary infection and infection; has obvious anti-inflammatory reaction to early inflammatory exudation and swelling, can accelerate the absorption of wound exudation, and promote the growth of epithelium and wound healing. In addition, the dispersible tablet of Callicarpa nudiflora has antiviral effect. Due to its wide pharmacological action, it is used for treating various skin diseases, such as acne vulgaris, herpes zoster, eczema, contact dermatitis, allergic purpura, cutaneous vasculitis, and after liquid nitrogen freezing operation.
Allergic purpura is also called self-limiting acute hemorrhage, is allergic vasculitis invading the skin and other organs, namely, the arterioles and capillaries, and the disease causes that in vivo IgA or IgG circulating immune complexes are formed due to pathogen infection, certain drug action, allergy and the like and are deposited on the capillaries on the upper layer of dermis to cause vasculitis. It is mainly manifested by purpura, abdominal pain, arthralgia and renal damage, but the blood platelets are not reduced.
The pathogenesis and mechanism of allergic purpura are not completely clarified so far, and may be related to streptococcus infection, virus infection, medicine, food, insect bite and the like, and the pathogenesis is that an immune complex is formed by combining an antigen and an antibody and is deposited on a blood vessel wall to activate complement, so that inflammation is generated on capillaries, small blood vessel walls and the periphery of the capillaries, the permeability of the blood vessel walls is increased, and various clinical manifestations are generated.
The callicarpa nudiflora is mainly distributed in Guangdong, Hainan, Guangxi provinces and other provinces of China by taking dry leaves as the medicine, and takes the five-finger mountain products in Hainan as the top grade. From the book of materia Medica of Tang, the newly added Chinese herbs were selected in the book of Chinese pharmacopoeia 2015. It is bitter, slightly pungent and neutral in flavor; collected all the year round, and root, stem and leaf can be used as medicine, and has the effects of stopping bleeding, diminishing inflammation, dissipating blood stasis, relieving swelling, resisting bacteria, removing toxicity and the like. As a clinical routine medicine, the callicarpa nudiflora is a single-medicine patent medicine and is suitable for hemostasis recovery and inflammation regression. Through acute toxicity test, the medicine is safe and nontoxic within a reasonable dosage concentration range. The chemical components found in Callicarpa nudiflora are mainly flavonoids, phenylpropanoids, iridoids and triterpenes, but the currently disclosed extraction method of Callicarpa nudiflora mainly adopts single-component organic solvent extraction.
For example, chinese patent application 201711285943.3 discloses a callicarpa nudiflora extract, its preparation method and application. The preparation method comprises the following steps: mixing Callicarpa nudiflora with an organic solvent, extracting to obtain an extracting solution, centrifuging, and collecting filtrate; the organic solvent is ethanol, the volume concentration of the ethanol is 50-70%, the content of total flavone in the prepared callicarpa nudiflora extract is at least 0.2mg/mL, the content of verbascoside is at least 2mg/mL, the mass percentage content of the verbascoside in the prepared callicarpa nudiflora extract is 2.6%, the extraction rate of active ingredients is low, and the requirement cannot be met.
For another example, chinese patent application 201910537780.6 discloses the use of beautyberry extract in preparing lipid-lowering drugs. The callicarpa nudiflora extract is prepared by the following method: the method comprises the steps of taking fresh and dried callicarpa nudiflora leaves, naturally drying, crushing and sieving, carrying out ultrasonic extraction by using methanol, cooling, freezing and centrifuging an extracting solution, standing, taking supernate to pass through a microporous filter membrane, and carrying out freeze drying on the filtrate to obtain a callicarpa nudiflora extract.
Therefore, it is necessary to develop a preparation method of callicarpa nudiflora extract which can effectively improve the extraction efficiency of callicarpa nudiflora and reduce the loss of effective components of traditional Chinese medicines.
Disclosure of Invention
Based on the defects and shortcomings in the prior art, one of the purposes of the invention is to provide a callicarpa nudiflora extract, which is extracted by using a multi-component organic solvent and controls the concentration (volume fraction) of an extraction solution, so that the content of active ingredients of a medicament can be effectively improved, the utilization rate of traditional Chinese medicine ingredients can be effectively improved, and the resource waste can be reduced.
In order to achieve the above purpose, the invention provides the following technical scheme:
on one hand, the invention provides a callicarpa nudiflora extract, which comprises flavonoids such as callicarpa pedunculata ketone, apigenin, verbascoside, luteolin and glycoside thereof.
On the other hand, the invention also provides an extraction method of the callicarpa nudiflora extract, which comprises the following steps:
s1, taking clean callicarpa nudiflora roots, stems and leaves, drying, crushing, adding water, ultrasonically soaking, and filtering to obtain filtrate 1 and filter residue 1 for later use;
s2, adding a mixed solvent into the filter residue 1 obtained in the step S1, and performing reflux extraction to obtain a filtrate 2 and a filter residue 2 for later use;
s3, mixing the filtrate 1 obtained in the step S1 and the filtrate 2 obtained in the step S2, filtering, and freeze-drying the filtrate to obtain the callicarpa nudiflora extract.
The crushing particle size in the step S1 is 100-150 meshes; preferably 120-150 mesh.
The ultrasonic frequency in the step S1 is 20 to 50KHz, preferably 30 to 40 KHz; the ultrasonic time is 20-30 minutes; preferably 25-30 minutes.
The mixed solvent in the step S2 is a mixed solution of ethanol, acetone and petroleum ether;
the volume ratio of the ethanol to the acetone to the petroleum ether is 1-5: 1-2: 1-2;
preferably, the volume ratio of ethanol, acetone and petroleum ether is 2-4: 1-2: 1-2;
more preferably, the volume ratio of ethanol, acetone and petroleum ether is 4: 1: 2.
the concentration of the ethanol is 60-80%, the concentration of the acetone is 50-60%, and the concentration of the petroleum ether is 50-60%.
Preferably, the concentration of the ethanol is 80%, the concentration of the acetone is 60%, and the concentration of the petroleum ether is 60%
The temperature of the reflux extraction in the above step S2 is 60 to 80 deg.C, preferably 60 to 70 deg.C.
The reflux extraction time in the step S2 is 5 to 10 minutes, and the reflux extraction times are 1 to 2 times.
The filtration in the step S3 is a microfiltration membrane filtration, and the particle size of the microfiltration membrane is 0.22 μm;
the mass volume ratio of the medicinal materials to the extraction solvent is 1-5 g: 10-50 mL;
preferably, the mass volume ratio of the medicinal materials to the extraction solvent is 1 g: 10mL, 2 g: 20mL, 3 g: 30mL, 4 g: 40mL or 5 g: 50 mL.
In some preferred embodiments, the present invention provides a method for extracting callicarpa nudiflora extract, comprising the steps of:
s1, taking clean callicarpa nudiflora roots, stems and leaves, drying, crushing to obtain particles with the particle size of 100-150 meshes, adding water, carrying out ultrasonic treatment under the condition of frequency of 20-50KHz for 20-30 minutes, and filtering to obtain filtrate 1 and filter residue 1 for later use;
s2, adding the filter residue 1 obtained in the step S1 into a mixture with the volume ratio of 1-5: 1-2: 1-2 times of reflux extraction in a mixed solvent of ethanol, acetone and petroleum ether at the temperature of 60-80 ℃ for 5-10 minutes, and obtaining filtrate 2 and filter residue 2 for later use;
s3, mixing the filtrate 1 obtained in the step S1 and the filtrate 2 obtained in the step S2, filtering the mixture through a 0.22 mu m microporous filter membrane, and freeze-drying the filtrate to obtain the callicarpa nudiflora extract.
The invention also provides application of the callicarpa nudiflora extract in preparation of a medicine for treating anaphylactoid purpura.
The invention also provides a composition which comprises the callicarpa nudiflora extract and pharmaceutically acceptable auxiliary materials.
The composition provided by the invention is in the dosage form of tablets, injections, capsules, granules, suppositories, ointments, creams or sprays, and preferably granules.
The invention also provides application of the composition in preparing a medicament for treating anaphylactoid purpura.
Compared with the prior art, the invention has the beneficial effects that:
(1) the extraction method of the callicarpa nudiflora extract provided by the invention adopts the mixed solvent for extraction, and can obviously improve the release of the active ingredients of the callicarpa nudiflora by controlling the volume ratio of the extraction solvent, so that the total extraction rate of flavonoids can be obviously improved;
(2) the invention can obviously improve the extraction efficiency and shorten the extraction time by controlling the concentration of the extraction solvent, can still ensure the efficient release of effective components at lower temperature, improve the extraction efficiency and reduce the waste of traditional Chinese medicines;
(3) the callicarpa nudiflora extract provided by the invention can be used for effectively treating anaphylactoid purpura, and is good in treatment effect, safe and effective.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
The callicarpa nudiflora used for the extraction test is a medicinal material purchased in a conventional pharmacy.
Example 1A method for extracting Callicarpa nudiflora extract
The method comprises the following steps:
s1, taking clean callicarpa nudiflora roots, stems and leaves, drying, crushing to 100 meshes, adding water, carrying out ultrasonic treatment under the condition of 20KHz frequency for 20 minutes, and filtering to obtain filtrate 1 and filter residue 1 for later use;
s2, adding the filter residue 1 obtained in the step S1 into a mixture with the volume ratio of 1: 1: 1, refluxing and extracting in a mixed solvent of 60% ethanol, 50% acetone and 50% petroleum ether at the temperature of 60 ℃ for 2 times of 10 minutes to obtain filtrate 2 and filter residue 2 for later use;
s3, mixing the filtrate 1 obtained in the step S1 and the filtrate 2 obtained in the step S2, filtering the mixture through a 0.22 mu m microporous filter membrane, and freeze-drying the filtrate to obtain the callicarpa nudiflora extract.
Example 2A method for extracting Callicarpa nudiflora extract
The method comprises the following steps:
s1, taking clean callicarpa nudiflora roots, stems and leaves, drying, crushing to 150 meshes, adding water, carrying out ultrasonic treatment under the condition of 50KHz frequency for 30 minutes, and filtering to obtain filtrate 1 and filter residue 1 for later use;
s2, adding the filter residue 1 obtained in the step S1 into a mixture with the volume ratio of 5: 2: 2, reflux extraction is carried out in a mixed solvent of 80% ethanol, 60% acetone and 60% petroleum ether, the temperature of reflux extraction is 80 ℃, the extraction time is 5 minutes, extraction is carried out for 2 times, and the obtained filtrate 2 and filter residue 2 are reserved;
s3, mixing the filtrate 1 obtained in the step S1 and the filtrate 2 obtained in the step S2, filtering the mixture through a 0.22 mu m microporous filter membrane, and freeze-drying the filtrate to obtain the callicarpa nudiflora extract.
Example 3A method for extracting Callicarpa nudiflora extract
The method comprises the following steps:
s1, taking clean callicarpa nudiflora roots, stems and leaves, drying, crushing to 120 meshes, adding water, carrying out ultrasonic treatment under the condition of 30KHz frequency for 25 minutes, and filtering to obtain filtrate 1 and filter residue 1 for later use;
s2, adding the filter residue 1 obtained in the step S1 into a mixture with the volume ratio of 4: 1: 2, reflux extraction is carried out in a mixed solvent of 80% ethanol, 60% acetone and 60% petroleum ether at the reflux extraction temperature of 70 ℃ for 8 minutes for 2 times, and filtrate 2 and filter residue 2 are obtained for later use;
s3, mixing the filtrate 1 obtained in the step S1 and the filtrate 2 obtained in the step S2, filtering the mixture through a 0.22 mu m microporous filter membrane, and freeze-drying the filtrate to obtain the callicarpa nudiflora extract.
Comparative example 1
The difference from example 3 is that the volume ratio of 80% ethanol, 60% acetone and 60% petroleum ether is 1: 5: 5, other operations and steps are the same as in example 3.
Comparative example 2
The difference from example 3 is that the volume ratio of 80% ethanol, 60% acetone and 60% petroleum ether is 10: 1: 1, other operations and steps are the same as in example 3.
Comparative example 3
The differences from example 3 are that the concentrations of ethanol, acetone and petroleum ether are 50%, 70% and 70%, respectively, and other operations and steps are the same as those of example 3.
Comparative example 4
The differences from example 3 are that the concentrations of ethanol, acetone and petroleum ether are 90%, 50% and 50%, respectively, and other operations and steps are the same as those of example 3.
Comparative example 5
The difference from example 3 is that only 70% ethanol is used and the other operations and steps are the same as example 3.
Comparative example 6
The difference from example 3 is that only 60% acetone is used and the other operations and steps are the same as example 3.
Test example 1 detection of extraction Rate
The extraction rates of verbascoside and luteolin and the total extraction rate of flavonoids in the callicarpa nudiflora extracts obtained in examples 1-3 and comparative examples 1-6 were measured.
The detection method comprises the following steps:
chromatographic conditions are as follows: octadecylsilane chemically bonded silica is used as filler, methanol-0.1% phosphoric acid solution (60: 40) is used as mobile phase, the detection wavelength is 348nm, and the number of theoretical plates is not less than 3000 calculated according to the peak of luteolin.
Octadecylsilane chemically bonded silica is used as filler, acetonitrile-0.1% acetic acid solution (16: 84) is used as mobile phase, the detection wavelength is 332nm, and the number of theoretical plates is not less than 4000 according to the peak of acteoside.
Preparation of a reference solution: weighing appropriate amount of luteolin and verbascoside as reference substances, and adding 70% methanol and 50% methanol respectively to obtain solutions containing 30mg and 40mg per ml.
Preparing a test solution:
and (3) respectively taking the samples of examples 1-3 and comparative examples 1-6, adding a solvent for dissolving, carrying out volume metering and filtering, and recording to obtain a test sample solution.
The determination method comprises the following steps: respectively sucking 10 μ L of reference solution and sample solution, injecting into liquid chromatograph, and measuring.
Specific extraction efficiencies are shown in tables 1-2 below.
TABLE 1 extraction rates of verbascoside and luteolin and total extraction rate of flavonoids from the callicarpa nudiflora extracts obtained in examples 1-3 and comparative examples 1-6
TABLE 2 concentrations of verbascoside, luteolin and flavonoids in the extracts prepared in examples 1-3 and comparative examples 1-6
The extracts prepared in examples 1 to 3 and comparative examples 1 to 6 were dissolved in water to obtain a liquid medicine, and the concentrations of verbascoside, luteolin and flavonoids in the liquid medicine were measured to obtain a liquid medicine concentration of 0.02 g/mL.
| Examples of the invention | Acteoside concentration | Luteolin concentration | Concentration of flavonoids |
| Example 1 | 3.2mg/mL | 0.18mg/mL | 0.29mg/mL |
| Example 2 | 3.4mg/mL | 0.20mg/mL | 0.28mg/mL |
| Example 3 | 3.7mg/mL | 0.24mg/mL | 0.31mg/mL |
| Comparative example 1 | 3.0mg/mL | 0.15mg/mL | 0.24mg/mL |
| Comparative example 2 | 2.9mg/mL | 0.13mg/mL | 0.23mg/mL |
| Comparative example 3 | 2.5mg/mL | 0.12mg/mL | 0.21mg/mL |
| Comparative example 4 | 2.6mg/mL | 0.12mg/mL | 0.22mg/mL |
| Comparative example 5 | 2.0mg/mL | 0.09mg/mL | 0.20mg/mL |
| Comparative example 6 | 2.1mg/mL | 0.10mg/mL | 0.20mg/mL |
Test II, animal test
The therapeutic effects of the extracts prepared in examples 1 to 3 and comparative examples 1 to 6 of the present invention on the mouse model of allergic purpura were studied.
Molding: randomly selecting 110 Kunming mice, female, age of the mice is 8-10 weeks, and body mass is 18-25g, and the mice are purchased from Guangzhou university of traditional Chinese medicine. After the test was started, 10 mice were randomly selected as a blank control group and fed with 5mmol/L HC1 solution, 0.5 ml/mouse, every other day for up to 10 weeks. The IgA nephropathy model was established for the remaining mice as follows: the tail vein is injected with 0.4mg/10g Indian ink 3 weeks before experiment, 1 time per week, and after 3 weeks, mice are fed with 0.1% gliadin, 5mmol/LHC1 solution, 0.5 ml/mouse, and fed every other day for 10 weeks. On the last 3 days of molding, tail vein injection was performed by adding 1mg of gliadin to 5mmol/L HC1 solution, pH7.4 Phosphate Buffered Saline (PBS), 0.2 ml/mouse per mouse, 1 time per day. 3d consecutively, starting at week 11, the model mice were randomly divided into model control and experimental groups of 10 mice each. The experimental mice were gavaged with 300mg/kg of the extract granules prepared in examples 1 to 3 and comparative examples 1 to 6 1 time per day for 3 weeks, and the blank control group and the model control group were each fed with distilled water (20ml/kg) of equal volume.
After 3 weeks of feeding, collecting urine for 6h for each group to perform quantitative determination of urine volume and urine protein; randomly selecting 5 mice in each group the next day, taking eyeballs, taking blood, centrifuging, separating serum, taking serum, and performing 24-hour urine protein quantification, urine erythrocyte count, serum creatinine, serum IL-4 content and IFN-gamma content measurement. The determination method comprises the following steps: 24h quantitative determination of urine protein: collecting urine volume for 6h by using a metabolism cage (one cage is placed for each 2, and 5 cages are placed for each group), detecting the urine volume after collecting the urine, and multiplying 4 times to obtain the total urine volume for 24 h; mouse urine protein quantification was determined by coomassie brilliant blue colorimetry. ② counting urine deformed red blood cells: measured using a phase contrast microscope. ③ measuring the IL-4 content in the serum: enzyme-linked immunosorbent assay is adopted. Measuring IFN-gamma content: enzyme-linked immunosorbent assay is adopted.
TABLE 3 quantitative comparison of urine volume and urine protein in 24 hours for each group of mice
| Examples of the invention | n | Quantitative urine protein (mg/24h) | Urine deformed erythrocyte (root/HP) |
| Blank control group | 10 | 0.304±0.132 | 0 |
| Model control group | 10 | 0.742±0.132 | 14.8±4.2 |
| EXAMPLE 1 group | 10 | 0.546±0.125** | 8.0±3.6** |
| EXAMPLE 2 group | 10 | 0.553±0.118** | 8.2±2.3** |
| EXAMPLE 3 group | 10 | 0.482±0.109** | 7.5±2.8** |
| Comparative example 1 group | 10 | 0.621±0.124* | 9.2±3.2* |
| Comparative example 2 group | 10 | 0.625±0.142* | 9.3±3.4* |
| Comparative example 3 group | 10 | 0.654±0.108* | 9.5±2.6* |
| Comparative example 4 group | 10 | 0.649±0.125* | 9.8±2.9* |
| Comparative example 5 group | 10 | 0.684±0.130* | 9.9±3.1* |
| Comparative example 6 group | 10 | 0.675±0.124* | 9.7±3.0* |
Note: compared to the model set: p is less than or equal to 0.01, P is less than or equal to 0.05
Compared with a blank group, the 24-hour urine protein quantification of the model group is obviously increased (P is less than or equal to 0.01), compared with the model group, the 24-hour urine protein quantification of the groups of examples 1 to 3 and the groups of comparative examples 1 to 6 is reduced to different degrees, the statistical significance is realized, and the 24-hour urine protein quantification of the groups of examples 1 to 3 is obviously reduced compared with the groups of comparative examples 1 to 6, and the statistical significance is realized. The urine examination of the blank group of mice does not find deformed red blood cells, and the rest groups have different numbers of deformed red blood cells, and compared with the blank group, the groups have significant difference, compared with the model group, the number of the denatured red blood cells of the groups of examples 1-3 is obviously reduced, and compared with the groups of comparative examples 1-6, the number of the denatured red blood cells is reduced, but is not obviously reduced compared with the groups of examples.
The results of serum creatinine, serum IL-4 levels, and IFN- γ levels for each group of mice are shown in Table 4.
TABLE 4
| Examples of the invention | n | Creatinine (mu mol/L) | Serum IL-4 content () | Serum IFN-gamma content |
| Blank control group | 10 | 48.6±9.6 | 0.58±0.12 | 50.23±2.56 |
| Model control group | 10 | 56.8±8.5 | 0.85±0.10 | 36.24±3.12 |
| EXAMPLE 1 group | 10 | 49.4±8.4 | 0.61±0.13** | 48.15±3.14** |
| EXAMPLE 2 group | 10 | 49.2±9.2 | 0.63±0.09** | 48.56±2.95** |
| EXAMPLE 3 group | 10 | 48.8±9.0 | 0.60±0.11** | 49.38±3.25** |
| Comparative example 1 group | 10 | 53.4±7.6 | 0.65±0.14* | 42.89±2.96* |
| Comparative example 2 group | 10 | 53.6±8.1 | 0.66±0.12* | 43.15±3.04* |
| Comparative example 3 group | 10 | 52.9±7.9 | 0.68±0.13* | 40.54±3.01* |
| Comparative example 4 group | 10 | 52.4±9.2 | 0.66±0.10* | 41.06±2.62* |
| Comparative example 5 group | 10 | 51.6±9.4 | 0.69±0.09* | 40.07±2.31* |
| Comparative example 6 group | 10 | 50.4±8.6 | 0.71±0.08* | 39.12±3.21* |
Note: compared to the model set: p is less than or equal to 0.01, P is less than or equal to 0.05
According to the detection results in the table 4, it can be seen that the serum creatinine changes are not obvious and are all within the normal range; compared with a blank group, the serum IFN-gamma content of the model group is obviously reduced, compared with the model group, the serum IFN-gamma content of the groups of examples 1-3 and the groups of comparative examples 1-6 can be increased to different degrees, and the statistical significance is achieved; compared with the blank group, the serum IL-4 content of the model group is obviously increased, and compared with the model group, the groups of examples 1-3 and the groups of comparative examples 1-6 can reduce the serum IL-4 content to different degrees, thereby having statistical significance.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (8)
1. A method for extracting a callicarpa nudiflora extract is characterized by comprising the following steps: the method comprises the following steps:
s1, taking clean callicarpa nudiflora roots, stems and leaves, drying, crushing, adding water, ultrasonically soaking, and filtering to obtain filtrate 1 and filter residue 1 for later use;
s2, adding a mixed solvent into the filter residue 1 obtained in the step S1, and performing reflux extraction to obtain a filtrate 2 and a filter residue 2 for later use;
s3, mixing the filtrate 1 obtained in the step S1 and the filtrate 2 obtained in the step S2, filtering, and freeze-drying the filtrate to obtain a callicarpa nudiflora extract;
the mixed solvent in the step S2 is a mixed solution of ethanol, acetone and petroleum ether; the volume ratio of the ethanol to the acetone to the petroleum ether is 1-5: 1-2: 1-2; the concentration of the ethanol is 60-80%, the concentration of the acetone is 50-60%, and the concentration of the petroleum ether is 50-60%.
2. The extraction method according to claim 1, characterized in that: the crushing particle size in the step S1 is 100-150 meshes.
3. The extraction method according to claim 1, characterized in that: the ultrasonic frequency in the step S1 is 20-50 KHz; the ultrasonic time is 20-30 minutes.
4. The extraction method according to claim 1, characterized in that: the volume ratio of the ethanol to the acetone to the petroleum ether is 4: 1: 2.
5. the extraction method according to claim 1, characterized in that: the concentration of the ethanol is 80%, the concentration of the acetone is 60%, and the concentration of the petroleum ether is 60%.
6. Use of the extract of callicarpa nudiflora obtained by the extraction method according to any one of claims 1-5 in the preparation of a medicament for treating allergic purpura.
7. A pharmaceutical composition for treating allergic purpura is characterized in that: comprises the callicarpa nudiflora extract obtained by the extraction method of any one of claims 1-5 and pharmaceutically acceptable auxiliary materials.
8. The pharmaceutical composition of claim 7, wherein: the dosage form of the pharmaceutical composition is tablets, injections, capsules, granules, suppositories, ointments, creams or sprays.
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| CN102205000A (en) * | 2010-10-15 | 2011-10-05 | 江西普正制药有限公司 | Callicarpa nudiflora Hook. et Arn. extract, preparation method and application thereof |
| CN106266262A (en) * | 2015-06-05 | 2017-01-04 | 九芝堂股份有限公司 | A kind of Callicarpa nudiflora extract with antiinflammatory action |
| CN106266263A (en) * | 2015-06-10 | 2017-01-04 | 九芝堂股份有限公司 | A kind of Chinese medicine composition and its production and use |
| CN108524685A (en) * | 2018-07-03 | 2018-09-14 | 江西普正制药有限公司 | A kind of composition that treating children's herpangina and its application |
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2020
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| CN102205000A (en) * | 2010-10-15 | 2011-10-05 | 江西普正制药有限公司 | Callicarpa nudiflora Hook. et Arn. extract, preparation method and application thereof |
| CN106266262A (en) * | 2015-06-05 | 2017-01-04 | 九芝堂股份有限公司 | A kind of Callicarpa nudiflora extract with antiinflammatory action |
| CN106266263A (en) * | 2015-06-10 | 2017-01-04 | 九芝堂股份有限公司 | A kind of Chinese medicine composition and its production and use |
| CN108524685A (en) * | 2018-07-03 | 2018-09-14 | 江西普正制药有限公司 | A kind of composition that treating children's herpangina and its application |
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