Application of folium artemisiae argyi polysaccharide in preparation of medicine for promoting wound healing

Abstract

The invention relates to the technical field of biological medicine, and discloses application of folium artemisiae argyi polysaccharide in preparation of a medicine for promoting wound healing. The folium artemisiae argyi polysaccharide is prepared according to the following steps: (1) performing steam distillation on folium artemisiae argyi, separating out a solid-phase product, and then drying the solid-phase product to obtain folium artemisiae argyi dregs; (2) mixing the folium artemisiae argyi dregs with water, performing ultrasonic extraction, and then filtering the obtained extract to obtain a folium artemisiae argyi crude dreg polysaccharide solution; (3) mixing the folium artemisiae argyi crude residue polysaccharide solution with a mixed solution of trichloromethane and n-butyl alcohol, and then standing for layering to obtain a deproteinized folium artemisiae argyi polysaccharide solution; and (4) mixing the deproteinized folium artemisiae argyi polysaccharide solution with ethanol, and carrying out alcohol precipitation. According to the technical scheme, the folium artemisiae argyi polysaccharide extracted from the folium artemisiae argyi dregs can be used for preparing the medicine for promoting wound healing, and meanwhile environmental pollution can be avoided.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to the application of Artemisia argyi polysaccharide in the preparation of drugs that promote wound healing. Background Technology

[0002] Artemisia argyi Levl. et Vant., a plant belonging to the genus Artemisia in the family Asteraceae, is a traditional Chinese medicine with a long history of medicinal use. It is recorded in classic medical books such as "Shennong's Classic of Materia Medica" and "Compendium of Materia Medica". It has the effects of warming the meridians and stopping bleeding, dispelling cold and relieving pain, and removing dampness and relieving itching. In modern clinical practice, it is widely used in the treatment of gynecological diseases, respiratory diseases and skin diseases.

[0003] Modern pharmacological studies have shown that Artemisia argyi contains various active ingredients such as volatile oils, polysaccharides, flavonoids, and terpenes. Among them, Artemisia argyi polysaccharides, as important water-soluble active ingredients, have significant anti-inflammatory, antibacterial, antioxidant, and immunomodulatory pharmacological effects. In the field of skin wound repair, uncontrolled inflammatory responses, insufficient cell proliferation and migration capacity, and abnormal tissue remodeling are key factors leading to delayed wound healing. Related reports indicate that polysaccharides can reduce wound inflammation by inhibiting the release of inflammatory factors (such as TNF-α and IL-6); at the same time, they can promote the proliferation and migration of epidermal cells (such as HaCaT cells) and fibroblasts, accelerate the formation of new granulation tissue and wound re-epithelialization, thereby promoting the wound healing process and demonstrating good potential for wound repair applications.

[0004] Currently, the application of mugwort in the medicinal and daily chemical fields mainly focuses on extracting volatile oils and flavonoids. The resulting residue is typically discarded as waste, leading to significant resource waste and potential environmental problems. However, research has found that even after extraction of volatile oils and flavonoids, the residue still contains a large amount of polysaccharides, which retain good biological activity. If polysaccharides can be efficiently extracted from mugwort residue, the entire industrial chain of mugwort resources can be utilized, reducing production costs and improving resource utilization.

[0005] Currently, research on Artemisia argyi polysaccharides mainly focuses on their direct extraction from fresh or dried Artemisia argyi leaves. Research on optimizing the extraction process of polysaccharides from Artemisia argyi residue and their application in the treatment of excisional wounds in mice has not been extensively conducted. Therefore, extracting polysaccharides from Artemisia argyi residue and exploring their effects on the healing of excisional wounds in mice has significant theoretical and practical value, and lays the foundation for the high-value utilization of Artemisia argyi resources and the development of novel wound repair products. Summary of the Invention

[0006] The purpose of this invention is to overcome the problems of resource waste and environmental pollution caused by the dregs of Artemisia argyi generated during the production process in existing technologies, and to provide an application of Artemisia argyi polysaccharide in the preparation of drugs that promote wound healing. According to the technical solution of this invention, Artemisia argyi polysaccharide extracted from Artemisia argyi dregs can be used to prepare drugs that promote wound healing, while also avoiding environmental pollution.

[0007] To achieve the above objectives, the present invention provides an application of Artemisia argyi polysaccharide in the preparation of a wound-healing drug, wherein the Artemisia argyi polysaccharide is prepared according to the following steps:

[0008] (1) The mugwort leaves are steam distilled and the solid product is separated. Then the solid product is dried to obtain mugwort residue.

[0009] (2) The mugwort residue and water are mixed and ultrasonically extracted, and then the resulting extract is filtered to obtain a mugwort crude residue polysaccharide solution.

[0010] (3) The Artemisia argyi crude residue polysaccharide solution and the mixed solution of chloroform and n-butanol are mixed and then allowed to stand and separate into layers to obtain a deproteinized Artemisia argyi polysaccharide solution.

[0011] (4) Mix the deproteinized Artemisia argyi polysaccharide solution with ethanol and perform alcohol precipitation.

[0012] Preferably, in the process of preparing a wound-healing drug using Artemisia argyi polysaccharide, the dosage form of the drug is at least one of ointment, tablet, pill, powder, decoction, granule, injection, capsule, and decocted paste.

[0013] Preferably, in step (1), the conditions for steam distillation include: a temperature of 80-160°C and a time of 2-10 hours.

[0014] Preferably, in step (1), the drying conditions include a temperature of 40-50°C and a time of 20-24 hours.

[0015] Preferably, in step (2), the conditions for ultrasonic extraction include: a temperature of 60-100℃, a time of 30-240 min, and a material-to-liquid ratio of 1g:(10-40)mL.

[0016] Preferably, in step (3), the volume ratio of chloroform to n-butanol in the mixed solution of chloroform and n-butanol is 3-6:1.

[0017] Preferably, in step (3), the volume ratio of the Artemisia argyi crude residue polysaccharide solution and the mixed solution of chloroform and n-butanol is 1:1-2.

[0018] Preferably, in step (3), the mixing time is 20-30 minutes.

[0019] Preferably, in step (4), the volume ratio of the deproteinized Artemisia argyi polysaccharide solution to the ethanol is 1:4-6.

[0020] Preferably, in step (4), the alcohol precipitation time is 10-12 hours.

[0021] According to the technical solution of the present invention, the polysaccharide extracted from the residue of Artemisia argyi can be used to prepare a drug that promotes wound healing, while avoiding environmental pollution. Attached Figure Description

[0022] Figure 1 The polysaccharide yield-time curves of Artemisia argyi polysaccharides extracted in Examples 1-5 of this invention are shown.

[0023] Figure 2 The polysaccharide yield-solid-liquid ratio curves of Artemisia argyi polysaccharides extracted in Examples 6-10 of this invention are shown.

[0024] Figure 3 The polysaccharide yield-extraction temperature curves of Artemisia argyi polysaccharides extracted in Examples 11-15 of this invention are shown below.

[0025] Figure 4 This is a curve showing the polysaccharide yield versus extraction number of the extracted Artemisia argyi polysaccharides from Examples 16-19 of this invention.

[0026] Figure 5 This is a comparison chart of wound healing rates in mice from the control group, positive control group, low-dose group, medium-dose group, and high-dose group in the test examples of this invention;

[0027] Figure 6 This is a schematic diagram showing the skin wound healing status of the control group, positive control group, low-dose group, medium-dose group, and high-dose group in the test examples of this invention;

[0028] Figure 7 The images show the HE staining results of mouse skin tissues from the control group, positive control group, low-dose group, medium-dose group, and high-dose group in the test examples of this invention.

[0029] Figure 8 This is a statistical chart of Masson-stained collagen fiber area in the control group, positive control group, low-dose group, medium-dose group, and high-dose group in the test examples of this invention;

[0030] Figure 9 The images show the results of Masson staining of mouse skin tissue in the control group, positive control group, low-dose group, medium-dose group, and high-dose group in the test examples of this invention. Detailed Implementation

[0031] The following provides a detailed description of specific embodiments of the present invention. It should be understood that the specific embodiments described herein are for illustrative and explanatory purposes only and are not intended to limit the scope of the invention.

[0032] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0033] This invention provides the application of Artemisia argyi polysaccharide in the preparation of drugs that promote wound healing, wherein the Artemisia argyi polysaccharide is prepared according to the following steps:

[0034] (1) The mugwort leaves are steam distilled and the solid product is separated. Then the solid product is dried to obtain mugwort residue.

[0035] (2) The mugwort residue and water are mixed and ultrasonically extracted, and then the resulting extract is filtered to obtain a mugwort crude residue polysaccharide solution.

[0036] (3) The Artemisia argyi crude residue polysaccharide solution and the mixed solution of chloroform and n-butanol are mixed and then allowed to stand and separate into layers to obtain a deproteinized Artemisia argyi polysaccharide solution.

[0037] (4) Mix the deproteinized Artemisia argyi polysaccharide solution with ethanol and perform alcohol precipitation.

[0038] In the process of preparing a wound-healing drug using Artemisia argyi polysaccharide, the dosage form of the drug can be at least one selected from ointment, tablet, pill, powder, decoction, granule, injection, capsule, and decocted paste. In the most preferred embodiment, the dosage form of the drug is an ointment.

[0039] In step (1), the conditions for steam distillation may include: a temperature of 80-160°C and a time of 2-10 hours. Preferably, the conditions for steam distillation include: a temperature of 90-100°C and a time of 6-8 hours. In a specific embodiment, the heating method for steam distillation is water bath heating. The steam distillation process can be carried out in an electrically heated constant-temperature water bath.

[0040] In step (1), the drying conditions may include a temperature of 40-50°C and a time of 20-24 hours. Preferably, the drying conditions include a temperature of 42-48°C and a time of 21-23 hours. The drying process can be carried out in an electrically heated forced-air drying oven.

[0041] In this invention, the mugwort leaves can be collected from the resource nursery of Hubei University of Traditional Chinese Medicine. The mugwort leaves are the dried leaves of Artemisia argyi, a plant in the Asteraceae family.

[0042] In step (2), the conditions for ultrasonic extraction may include: a temperature of 60-100℃, a time of 30-240 min, and a solid-liquid ratio of 1 g:(10-40) mL. Preferably, the conditions for ultrasonic extraction include: a temperature of 70-90℃, a time of 50-220 min, and a solid-liquid ratio of 1 g:(15-35) mL.

[0043] In step (2), the ultrasonic extraction can be performed once or multiple times, preferably multiple times.

[0044] In step (3), the volume ratio of chloroform to n-butanol in the mixed solution of chloroform and n-butanol can be 3-6:1. In the most preferred embodiment, the volume ratio of chloroform to n-butanol is 4:1.

[0045] In step (3), the volume ratio of the Artemisia argyi crude residue polysaccharide solution and the mixed solution of chloroform and n-butanol can be 1:1-2, preferably 1:1-1.5.

[0046] In step (3), the mixing time can be 20-30 min, preferably 26-30 min.

[0047] In step (4), the volume ratio of the deproteinized Artemisia argyi polysaccharide solution to the ethanol can be 1:4-6, preferably 1:5-5.5.

[0048] In step (4), the alcohol precipitation time can be 10-12 hours, preferably 10.5-11.5 hours.

[0049] The following examples further illustrate the application of Artemisia argyi polysaccharide in the preparation of wound-healing drugs according to the present invention. These examples are implemented based on the technical solution of the present invention, providing detailed implementation methods and specific operating procedures; however, the scope of protection of the present invention is not limited to the following examples.

[0050] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods in the art. Unless otherwise specified, the experimental materials used in the following embodiments are commercially available.

[0051] Example 1

[0052] (1) 1g of Artemisia argyi leaves (collected from the resource garden of Hubei University of Traditional Chinese Medicine, dried leaves of Artemisia argyi, belonging to the Asteraceae family, the same below) were placed in a distillation apparatus and steam distilled at 100℃ for 6 hours. The remaining solid product was collected and placed in an electric heating drying oven and dried at 45℃ for 22 hours to obtain Artemisia argyi residue.

[0053] (2) The Artemisia argyi residue obtained in step (1) is mixed with water and subjected to ultrasonic extraction. The conditions for ultrasonic extraction include: temperature of 90℃, time of 30min, material-liquid ratio of 1g:25mL, extraction once. The obtained extract is then filtered to obtain Artemisia argyi crude residue polysaccharide solution.

[0054] (3) The Artemisia argyi crude residue polysaccharide solution obtained in step (2) and the mixed solution of chloroform and n-butanol (the volume ratio of chloroform and n-butanol is 1:4) are mixed at a volume ratio of 1:1, shaken for 30 minutes and allowed to stand to separate into layers to obtain the deproteinized Artemisia argyi polysaccharide solution.

[0055] (4) The deproteinized Artemisia argyi polysaccharide solution obtained in step (3) and 95% ethanol were mixed at a volume ratio of 1:5 and subjected to alcohol precipitation for 11 hours to obtain Artemisia argyi polysaccharide A1.

[0056] Example 2

[0057] Artemisia argyi polysaccharide was prepared according to the method in Example 1, except that in step (2), the extraction time was adjusted to 60 min to obtain Artemisia argyi polysaccharide A2.

[0058] Example 3

[0059] Artemisia argyi polysaccharide was prepared according to the method of Example 1, except that in step (2), the extraction time was adjusted to 120 min to obtain Artemisia argyi polysaccharide A3.

[0060] Example 4

[0061] Artemisia argyi polysaccharide was prepared according to the method in Example 1, except that in step (2), the extraction time was adjusted to 180 min to obtain Artemisia argyi polysaccharide A4.

[0062] Example 5

[0063] Artemisia argyi polysaccharide was prepared according to the method in Example 1, except that in step (2), the extraction time was adjusted to 240 min to obtain Artemisia argyi polysaccharide A5.

[0064] Example 6

[0065] (1) Place 1g of Artemisia argyi in a distillation apparatus and steam distill at 120℃ for 8 hours. Collect the remaining solid product and dry it in an electric heating drying oven at 48℃ for 22 hours to obtain Artemisia argyi residue.

[0066] (2) The mugwort residue obtained in step (1) is mixed with water and subjected to ultrasonic extraction. The conditions for ultrasonic extraction include: temperature of 90℃, time of 60min, material-liquid ratio of 1g:10mL, extraction once. The extracted extract is then filtered to obtain a mugwort crude residue polysaccharide solution.

[0067] (3) The Artemisia argyi crude residue polysaccharide solution obtained in step (2) and the mixed solution of chloroform and n-butanol (the volume ratio of chloroform and n-butanol is 1:4) are mixed at a volume ratio of 1:1, shaken for 30 minutes and allowed to stand to separate into layers to obtain the deproteinized Artemisia argyi polysaccharide solution.

[0068] (4) The deproteinized Artemisia argyi polysaccharide solution obtained in step (3) and 95% ethanol were mixed at a volume ratio of 1:5 and subjected to alcohol precipitation for 12 hours to obtain Artemisia argyi polysaccharide A6.

[0069] Example 7

[0070] Artemisia argyi polysaccharide was prepared according to the method of Example 6, except that in step (2), the material-liquid ratio was adjusted to 1g:20mL to obtain Artemisia argyi polysaccharide A7.

[0071] Example 8

[0072] Artemisia argyi polysaccharide was prepared according to the method of Example 6, except that in step (2), the material-liquid ratio was adjusted to 1g:25mL to obtain Artemisia argyi polysaccharide A8.

[0073] Example 9

[0074] Artemisia argyi polysaccharide was prepared according to the method of Example 6, except that in step (2), the material-liquid ratio was adjusted to 1g:30mL to obtain Artemisia argyi polysaccharide A9.

[0075] Example 10

[0076] Artemisia argyi polysaccharide was prepared according to the method of Example 6, except that in step (2), the material-liquid ratio was adjusted to 1g:40mL to obtain Artemisia argyi polysaccharide A10.

[0077] Example 11

[0078] (1) Place 1g of Artemisia argyi in a distillation apparatus and steam distill at 100℃ for 5 hours. Collect the remaining solid product and dry it in an electric heating drying oven at 43℃ for 24 hours to obtain Artemisia argyi residue.

[0079] (2) The mugwort residue obtained in step (1) is mixed with water and subjected to ultrasonic extraction. The conditions for ultrasonic extraction include: temperature of 60℃, time of 60min, material-liquid ratio of 1g:20mL, extraction once. The obtained extract is then filtered to obtain mugwort crude residue polysaccharide solution.

[0080] (3) The Artemisia argyi crude residue polysaccharide solution obtained in step (2) and the mixed solution of chloroform and n-butanol (the volume ratio of chloroform and n-butanol is 1:4) are mixed at a volume ratio of 1:1, shaken for 30 minutes and allowed to stand to separate into layers to obtain the deproteinized Artemisia argyi polysaccharide solution.

[0081] (4) The deproteinized Artemisia argyi polysaccharide solution obtained in step (3) and 95% ethanol at a volume ratio of 1:5 were mixed and precipitated for 12 hours to obtain Artemisia argyi polysaccharide A11.

[0082] Example 12

[0083] Artemisia argyi polysaccharide was prepared according to the method of Example 11, except that in step (2), the extraction temperature was adjusted to 70°C to obtain Artemisia argyi polysaccharide A12.

[0084] Example 13

[0085] Artemisia argyi polysaccharide was prepared according to the method of Example 11, except that in step (2), the extraction temperature was adjusted to 80°C to obtain Artemisia argyi polysaccharide A13.

[0086] Example 14

[0087] Artemisia argyi polysaccharide was prepared according to the method of Example 11, except that in step (2), the extraction temperature was adjusted to 90°C to obtain Artemisia argyi polysaccharide A14.

[0088] Example 15

[0089] Artemisia argyi polysaccharide was prepared according to the method of Example 11, except that in step (2), the extraction temperature was adjusted to 100°C to obtain Artemisia argyi polysaccharide A15.

[0090] Example 16

[0091] (1) Place 1g of Artemisia argyi in a distillation apparatus and steam distill at 100℃ for 6 hours. Collect the remaining solid product and dry it in an electric heating drying oven at 45℃ for 22 hours to obtain Artemisia argyi residue.

[0092] (2) The mugwort residue obtained in step (1) is mixed with water and subjected to ultrasonic extraction. The conditions for ultrasonic extraction include: temperature of 90℃, time of 60min, material-liquid ratio of 1g:25mL, extraction once. The obtained extract is then filtered to obtain mugwort crude residue polysaccharide solution.

[0093] (3) The Artemisia argyi crude residue polysaccharide solution obtained in step (2) and the mixed solution of chloroform and n-butanol (the volume ratio of chloroform and n-butanol is 1:4) are mixed at a volume ratio of 1:1, shaken for 30 minutes and allowed to stand to separate into layers to obtain the deproteinized Artemisia argyi polysaccharide solution.

[0094] (4) The deproteinized Artemisia argyi polysaccharide solution obtained in step (3) and 95% ethanol were mixed at a volume ratio of 1:4 and subjected to alcohol precipitation for 10 hours to obtain Artemisia argyi polysaccharide A16.

[0095] Example 17

[0096] Artemisia argyi polysaccharide was prepared according to the method of Example 16, except that in step (2), the number of extractions was adjusted to 2 times to obtain Artemisia argyi polysaccharide A17.

[0097] Example 18

[0098] Artemisia argyi polysaccharide was prepared according to the method of Example 16, except that in step (2), the number of extractions was adjusted to 3 times to obtain Artemisia argyi polysaccharide A18.

[0099] Example 19

[0100] Artemisia argyi polysaccharide was prepared according to the method of Example 16, except that in step (2), the number of extractions was adjusted to 4 times to obtain Artemisia argyi polysaccharide A19.

[0101] Test case

[0102] (1) Determination of polysaccharide content: Glucose solutions with concentrations of 10, 40, 80, 120, and 160 μg / mL were prepared, with distilled water as the blank group. 200 μL of each solution was pipetted into a 50 mL centrifuge tube, followed by 5 mL of concentrated sulfuric acid and 1 mL of 5% phenol. The solutions were allowed to stand for 5 min, then boiled in a water bath for 15 min. After cooling to room temperature, the absorbance at 490 nm was measured using a microplate reader (Bio-Tek Synergy2, Gene Biosciences, USA). A standard curve and regression equation were plotted with polysaccharide concentration on the x-axis and absorbance on the y-axis. The standard curve equation was: Y = 0.0025x + 0.0801, R0 = 0.0025x + 0.0801. 2 =0.9966. The formula for calculating polysaccharide content is as follows:

[0103] Polysaccharide content = (C × D / W) × 100%

[0104] In the formula: C is the glucose concentration of the sample (μg / mL); D is the dilution factor of the sample; W is the concentration of the sample (μg / mL).

[0105] The polysaccharide yield can be calculated based on the obtained polysaccharide content.

[0106] Figure 1 The polysaccharide yield-time curves of Artemisia argyi polysaccharides A1-A5 extracted in Examples 1-5 are shown. Figure 1 It was observed that the polysaccharide yield gradually increased with increasing extraction time, reaching 11.17 mg / g after 1 hour of extraction. The yield then began to decrease after 1 hour. This is likely because polysaccharides require time to diffuse from the powder into the water, resulting in a lower polysaccharide concentration in the water during the first hour of extraction. After 1 hour, the polysaccharides in the water reached saturation, and further extraction time no longer increased the polysaccharide yield. Furthermore, the increased extraction time may alter the polysaccharide structure, leading to a decrease in yield.

[0107] Figure 2 The diagram shows the polysaccharide yield-solid-liquid ratio curves of Artemisia argyi polysaccharides A1-A6 extracted in Examples 6-10. Figure 2 It was observed that the yield of Artemisia argyi polysaccharides initially increased and then decreased with increasing material-to-liquid ratio, reaching a maximum of 9.16 mg / g when the ratio reached 1 g:25 mL. This result is likely due to the increased concentration difference between the intracellular and extracellular fluids of Artemisia argyi, accelerating the diffusion of the solution into the cells and thus increasing the amount of polysaccharides dissolved, resulting in a higher yield. However, excessively high water content during extraction leads to a diverse and mixed composition of dissolved substances. These impurities may affect the effective release and utilization of polysaccharides, impacting the overall yield.

[0108] Figure 3 The polysaccharide yield-extraction temperature curves of Artemisia argyi polysaccharides A11-A15 extracted in Examples 11-15 are shown. Figure 3 As shown, the extraction yield of Artemisia argyi polysaccharides initially increased and then decreased with increasing extraction temperature. The extraction rate reached its maximum at 90℃, with a yield of 13.53 mg / mg. The extraction rate peaked at 90℃ and then gradually decreased. This may be because higher temperatures accelerate solvent molecule movement, leading to more efficient dissolution of polysaccharides. However, as the temperature rises above the optimum, the polysaccharide structure is damaged. Polysaccharides are macromolecules, and high temperatures cause the glycosidic bonds to break, resulting in degradation. The degraded smaller molecule fragments may no longer belong to the target polysaccharide or have altered solubility, leading to a decrease in extraction rate.

[0109] Figure 4 The diagram shows the polysaccharide yield-extraction number curves of Artemisia argyi polysaccharides A16-A19 extracted in Examples 16-19. Figure 4 It can be seen that the polysaccharide yield gradually increases with the number of extractions of Artemisia argyi polysaccharide, but the increase is not significant. When the number of extractions reaches 3, the yield of Artemisia argyi polysaccharide reaches 11.24 mg / g. After the number of extractions reaches 3, the polysaccharide yield decreases slightly. This may be because as the number of extractions increases, the solvent can quickly penetrate the raw material and preferentially dissolve a large amount of easily soluble free polysaccharides. At this time, each extraction can obtain a higher single extraction yield, and the total extraction rate increases accordingly. As the number of extractions increases, impurities such as proteins, tannins, and pigments in the raw material will continue to dissolve, and their content will gradually increase. Impurities may combine with polysaccharides to form insoluble complexes, such as polysaccharide-protein complexes and polysaccharide-tannin precipitates, which cannot be detected, resulting in a decrease in the calculated extraction rate.

[0110] (2) Evaluation of wound healing efficacy: Male ICR mice weighing 5-6 weeks (18-22g) were selected and fed at 22-25℃. After one week of observation without mortality or mental abnormalities, the mice were randomly divided into 5 groups of 6 mice each. After hair removal from the back, the mice were anesthetized by intraperitoneal injection of 0.375% sodium pentobarbital solution (0.2mL / 10g). A round hole was made in the back of the mouse, and a full-thickness skin with a diameter of 10.0mm was excised. After hemostasis, the skin was bandaged with sterile gauze. The control group was given petrolatum without polysaccharides, the positive control group was given 5000 IU / mouse / d recombinant bovine basic fibroblast growth factor gel, and the low, medium, and high dose groups of Artemisia argyi polysaccharides were given 10mg / mouse / d, 20mg / mouse / d, and 40mg / mouse / d, respectively, for 2 consecutive weeks. The wound healing of the mice was photographed and the mice were weighed daily. After administration, the mice were euthanized, and the skin tissue from the wound was cut off with sterile scissors and fixed in 4% paraformaldehyde solution at 4°C for histopathological sectioning and staining.

[0111] Figure 5 The skin healing rates of mouse wounds are shown in the control group, positive control group, low-dose group, medium-dose group, and high-dose group. Figure 6 The study showed the wound healing of mice in the control group, positive control group, low-dose group, medium-dose group, and high-dose group. Compared with the control group, the wounds in the positive control group and the medium-dose and high-dose groups of Artemisia argyi polysaccharide showed different degrees of healing, with significant wound healing (P<0.05 or P<0.01). The results indicate that Artemisia argyi polysaccharide can promote the healing of surgical wounds in mice.

[0112] Figure 7The results of HE staining of mouse skin tissue from the control group, positive control group, low-dose group, medium-dose group, and high-dose group are shown. In the control group, the epidermis of the back skin tissue of mice showed significant thickening, hyperkeratosis, and downward extension of epidermal ridges, with extensive inflammatory cell infiltration in the dermis. In the low-dose Artemisia argyi polysaccharide group, the epidermis was slightly thickened, with Munro microabscesses forming in the stratum corneum and partial inflammatory cell infiltration in the dermis. In the positive control group and the medium- and high-dose Artemisia argyi polysaccharide groups, the epidermal thickness approached normal, with a small amount of inflammatory cell infiltration in the dermis, significantly reduced compared to the model group. These results indicate that Artemisia argyi polysaccharide can improve the condition of skin damage from a histopathological perspective.

[0113] Figure 8 The following is a statistical chart showing the area of ​​Masson-stained collagen fibers in the control group, positive control group, low-dose group, medium-dose group, and high-dose group. Figure 9 The results of Masson staining are shown for the control group, positive control group, low-dose group, medium-dose group, and high-dose group. Masson staining was used to observe the collagen fiber formation in the skin tissue of each group. Compared with the control group, the area of ​​collagen fibers in the low, medium, and high-dose groups of Artemisia argyi polysaccharide was increased, indicating that Artemisia argyi polysaccharide has the effect of promoting collagen fiber formation, thereby promoting wound healing in mice.

[0114] As can be seen from the results of the above embodiments, according to the technical solution of the present invention, the polysaccharide extracted from the residue of Artemisia argyi can be used to prepare a drug that promotes wound healing, while avoiding environmental pollution.

[0115] The preferred embodiments of the present invention have been described in detail above; however, the present invention is not limited thereto. Within the scope of the inventive concept, various simple modifications can be made to the technical solutions of the present invention, including combinations of various technical features in any other suitable manner. These simple modifications and combinations should also be considered as the content disclosed in the present invention and are all within the protection scope of the present invention.

Claims

1. The application of Artemisia argyi polysaccharide in the preparation of wound-healing drugs, characterized in that, The Artemisia argyi polysaccharide was prepared according to the following steps: (1) The mugwort leaves are steam distilled and the solid product is separated. Then the solid product is dried to obtain mugwort residue. (2) The mugwort residue and water are mixed and ultrasonically extracted, and then the resulting extract is filtered to obtain a mugwort crude residue polysaccharide solution. (3) The Artemisia argyi crude residue polysaccharide solution and the mixed solution of chloroform and n-butanol are mixed and then allowed to stand and separate into layers to obtain a deproteinized Artemisia argyi polysaccharide solution. (4) Mix the deproteinized Artemisia argyi polysaccharide solution with ethanol and perform alcohol precipitation.

2. The application according to claim 1, characterized in that, In the process of preparing a wound-healing drug using Artemisia argyi polysaccharide, the dosage form of the drug is at least one of ointment, tablet, pill, powder, decoction, granule, injection, capsule and decoction paste.

3. The application according to claim 1, characterized in that, In step (1), the conditions for steam distillation include: a temperature of 80-160℃ and a time of 2-10h.

4. The application according to claim 1 or 3, characterized in that, In step (1), the drying conditions include a temperature of 40-50°C and a time of 20-24 hours.

5. The application according to claim 1, characterized in that, In step (2), the conditions for ultrasonic extraction include: a temperature of 60-100℃, a time of 30-240 min, and a material-to-liquid ratio of 1g:(10-40)mL.

6. The application according to claim 1, characterized in that, In step (3), the volume ratio of chloroform to n-butanol in the mixed solution of chloroform and n-butanol is 3-6:

1.

7. The application according to claim 1 or 6, characterized in that, In step (3), the volume ratio of the Artemisia argyi crude residue polysaccharide solution and the mixed solution of chloroform and n-butanol is 1:1-2.

8. The application according to claim 1, 6, or 7, characterized in that, In step (3), the mixing time is 20-30 minutes.

9. The application according to claim 1, characterized in that, In step (4), the volume ratio of the deproteinized Artemisia argyi polysaccharide solution to the ethanol is 1:4-6.

10. The application according to claim 1 or 9, characterized in that, In step (4), the alcohol precipitation time is 10-12 hours.

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